Abstract
Sulforaphane is a natural compound with neuroprotective activity, but its effects on hypothalamus remain unknown. In line with this, astrocytes are critical cells to maintain brain homeostasis, and hypothalamic astrocytes are fundamental for sensing and responding to environmental changes involved in a variety of homeostatic functions. Changes in brain functionality, particularly associated with hypothalamic astrocytes, can contribute to age-related neurochemical alterations and, consequently, neurodegenerative diseases. Thus, here, we investigated the glioprotective effects of sulforaphane on hypothalamic astrocyte cultures and hypothalamic cell suspension obtained from aged Wistar rats (24 months old). Sulforaphane showed anti-inflammatory and antioxidant properties, as well as modulated the mRNA expression of astroglial markers, such as aldehyde dehydrogenase 1 family member L1, aquaporin 4, and vascular endothelial growth factor. In addition, it increased the expression and extracellular levels of trophic factors, such as glia-derived neurotrophic factor and nerve growth factor, as well as the release of brain-derived neurotrophic factor and the mRNA of TrkA, which is a receptor associated with trophic factors. Sulforaphane also modulated the expression of classical pathways associated with glioprotection, including nuclear factor erythroid-derived 2-like 2, heme oxygenase-1, nuclear factor kappa B p65 subunit, and AMP-activated protein kinase. Finally, a cell suspension with neurons and glial cells was used to confirm the predominant effect of sulforaphane in glial cells. In summary, this study indicated the anti-aging and glioprotective activities of sulforaphane in aged astrocytes.
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Introduction
The hypothalamus is a crucial brain region that regulates energy balance, body temperature, circadian rhythm, reproduction, as well as vision and emotions [1,2,3,4,5,6,7,8]. It acts as a bridge between the central nervous system (CNS) and the periphery of the body by receiving information and generating an appropriate response. Evidence suggests that the hypothalamus actively participates in the aging process and can be a target for development of anti-aging and therapeutical strategies [9, 10]. In this sense, aging is a biological process that leads to a progressive decline in physical and cognitive abilities, resulting in impaired adaptive responses to environmental stressors. In the CNS, aging changes several neural functions, and hypothalamic astrocytes can contribute to age-related neurochemical alterations [11,12,13].
Astrocytes are a type of glial cells that play multiple roles in maintaining brain homeostasis. They regulate synaptic plasticity, neurotransmitter metabolism, and ionic balance; provide metabolic support to neurons; produce and release growth factors, inflammatory mediators, and antioxidants; and maintain the blood–brain barrier [11, 14,15,16,17,18]. In particular, hypothalamic astrocytes are fundamental for sensing and responding to environmental changes since they have receptors and transporters for hormones and molecules involved in a variety of homeostatic functions [19,20,21,22,23]. To promote an overall improvement of the CNS functions, glioprotection relies on specific responses of glial cells to protect themselves and neighbouring cells from damage, and it can be achieved by using specific molecules [17].
Sulforaphane is a naturally occurring compound found in cruciferous vegetables, such as broccoli, kale, and Brussels sprouts as a system of defense against pathogen attack [24, 25]. It has antioxidant and anti-inflammatory properties and has been shown to have several health benefits, including the improvement of cardiovascular health [17, 25,26,27,28,29,30]. The concentration of sulforaphane in these dietary sources varies depending on the type of cruciferous vegetable and even on the degree of maturation [31]. In rats, after oral administration of 50 μmol sulforaphane, the peak of plasma concentration reaches 20 μM after 4 h, a concentration that is relevant in in vitro cell studies [32]. Additionally, sulforaphane can cross the blood–brain barrier and reach the CNS to exert neuroprotective effects in pathological conditions such as stroke, traumatic brain injury, and neurodegenerative diseases [33,34,35]. However, the effects of sulforaphane on aged astrocytes remain unclear.
In this context, the purpose of the present study was to characterize the glioprotective effects of sulforaphane in the hypothalamus of aged rats, focused on astrocytes. For this, we evaluated neurochemical parameters related to inflammation and trophic factors (extracellular content and mRNA expression), as well as mRNA expression of genes associated with antioxidant activity/redox homeostasis, cytoprotective responses, astroglial and senescence markers in hypothalamic astrocyte cultures obtained from aged Wistar rats. In addition, to confirm the predominant effect of sulforaphane on glial cells, we evaluated the expression of the same parameters measured in cultured astrocytes in a cellular suspension of hypothalamic tissue containing both neurons and glial cells obtained from aged Wistar rats. To our knowledge, this is the first study that explores the anti-aging and glioprotective activities of sulforaphane in aged astrocytes.
Materials and Methods
Reagents
Dulbecco’s modified Eagle’s medium/F12 (DMEM/F12), TRIzol Reagent, other materials for cell culture, ELISA kits for interleukin-1β (IL-1β), interleukin-6 (IL-6), interleukin-10 (IL-10), BDNF, nerve growth factor brain-derived neurotrophic factor (BDNF) and tumor necrosis factor-ɑ (TNF-ɑ) were purchased from Gibco/Invitrogen (Carlsbad, CA, USA). Poly-l-lysine, sulforaphane and methylthiazolyldiphenyl-tetrazolium bromide (MTT) were purchased from Sigma–Aldrich (St. Louis, MO, USA). ELISA kit for glia-derived neurotrophic factor (GDNF) was purchased from R&D Systems (Minneapolis, MN, USA). High Capacity cDNA Reverse Transcription Kit, TaqMan real-time RT-PCR system, primers and probes were purchased from Applied Biosystems (Thermo Fisher Scientific).
All other chemicals were from common commercial suppliers.
Animals
Male Wistar rats (24 months old) were obtained from the breeding colony of Department of Biochemistry (Federal University of Rio Grande do Sul, Porto Alegre, Brazil) and maintained under a controlled environment (12 h-light/12 h-dark cycle, 22 ± 1 °C; ad libitum access to food and water). The animals received regular laboratory chow (Nuvilab-CR1, from Nuvital, Brazil). All animal experiments were performed in accordance with the National Institute of Health (NIH) Guide for the Care and Use of Laboratory Animals and were approved by the Federal University of Rio Grande do Sul Animal Care and Use Committee (process number 35387).
Primary Hypothalamic Astrocyte Cultures from Aged Rats
The protocol described previously by Santos et al. [11] was used. Wistar rats at 24 months old had their hypothalamus dissected, and the meninges removed. The tissue was enzymatically digested in Hank’s balanced salt solution (HBSS) containing 0.05% trypsin at 37 °C for 7 min. The tissue was then mechanically dissociated for 7 min and centrifuged at 100×g for 5 min. The pellet was resuspended in HBSS and again mechanically dissociated until complete homogenization, and then centrifuged at 100×g for 5 min. Then, cells were resuspended in DMEM/F12, supplemented with 10% fetal bovine serum (FBS), 15 mM HEPES, 14.3 mM NaHCO3, 2.5 μg/mL amphotericin B, and 0.05 mg/mL gentamicin. Cells were seeded (approximately 2–4 × 105 cells/cm2) into 6-well plates pre-coated with poly-l-lysine and cultured at 37 °C in a 5% CO2 incubator. After 24 h, the culture medium was exchanged; during the first week, the medium was replaced once every 2 days, and from the second week on, once every 4 days. From the second week on, the astrocytes received medium supplemented with 20% FBS until they reached confluence (at approximately the fourth week). No dibutyryl-cAMP was added to the culture medium. To determine whether the culture contained microglia or neurons after reaching confluence, we used anti-ß-tubulin III, anti-NeuN, and anti-CD11, and less than 5% of cells were labeled [11].
Hypothalamic Cell Suspension from Aged Rats
In addition to the primary culture, a cell suspension from the hypothalamic tissue was prepared. The hypothalamus from 24 months old Wistar rats were enzymatically digested in HBSS containing 0.05% trypsin, followed by a mechanical dissociation and centrifugation 100×g for 5 min. Cells were resuspended in serum-free DMEM/F12 and immediately incubated with sulforaphane.
Sulforaphane Treatments
To evaluate the effect of sulforaphane on hypothalamic astrocyte cultures, the cells were incubated in the absence (control conditions) or presence of 5 µM sulforaphane dissolved in dimethyl sulfoxide (DMSO) during 24 h at 37 °C in an atmosphere with 5% CO2 in serum-free DMEM/F12 medium [36]. The hypothalamic cell suspension was incubated in the presence or absence of 5 µM sulforaphane dissolved in DMSO during 1 h at 37 °C in an atmosphere with 5% CO2 in serum-free DMEM/F12 medium. It is noteworthy that a shorter time of incubation with sulforaphane was used in cell suspension from hypothalamus due to the viability of this tissue preparation. It should be also noted that the final concentration of DMSO (used as vehicle) did not present any effect on astroglial cells neither hypothalamic tissue [36, 37]. The concentration of sulforaphane utilized in this study is readily achievable in rat plasma after a single dose of sulforaphane [32, 38].
Cell Viability and Membrane Integrity Assays
The viability of astrocyte cultures following sulforaphane treatment was determined by performing the MTT reduction assay. MTT was added at a final concentration of 50 µg/mL and incubated for 3 h at 37 °C in a 5% CO2 atmosphere. After removing the medium, the MTT crystals were dissolved in DMSO, and the absorbance was measured at 560 and 650 nm [39].
Furthermore, the integrity of the cell membrane was evaluated using the propidium iodide (PI) incorporation assay. Astrocyte cultures were incubated with 7.5 µM PI for 30 min before the end of the sulforaphane treatment at 37 °C in an atmosphere with 5% CO2. Loss of membrane integrity results in fluorescent nuclei labeling with PI, which was quantified using OptiQuant software (Packard Instrument Company).
Trophic Factors and Inflammatory Response Measurements
BDNF, GDNF, and NGF levels were measured in the extracellular medium of astrocyte cultures, using commercial ELISA kits. The results are expressed in pg/mL. Cytokine levels were measured in the extracellular medium of astrocyte cultures using ELISA kits for TNF-ɑ, IL-1β, IL-6, and IL-10 and the results are expressed in pg/mL.
RNA Extraction and Quantitative RT-PCR
Total RNA was isolated from astrocyte cultures or hypothalamic cell suspension using TRIzol Reagent. Extracted RNA (1 μg) was submitted to cDNA synthesis by High-Capacity cDNA Reverse Transcription Kit. Quantitative PCR determination of the mRNAs encoding adenosine receptors A1 (#Rn00567668_m1), A2A (#Rn00583935_m1), and A2B (#Rn00567697_m1), aldehyde dehydrogenase 1 family member L1 (ALDH1L1) (#Rn00674034_m1), AMP-activated protein kinase (AMPK) (#Rn00576935_m1), aquaporin 4 (AQP4) (#Rn00563196_m1), β-actin (#Rn00667869_m1), BDNF (#Rn02531967_s1), cyclooxygenase 2 (COX-2; #Rn01483828_m1), glutamate-cysteine ligase (GCL; #Rn00689046_ m1), GDNF (#Rn07311775_m1), glial fibrillary acidic protein (GFAP; #Rn00566603_m1), glutamine synthetase (GS; #Rn01483107_m1), heme oxygenase-1 (HO-1) (#Rn01536933_ m1), IL-1β (#Rn00580432_m1), IL-6 (#Rn01410330_m1), IL-10 (#Rn00563409_m1), inducible nitric oxide synthase (iNOS; #Rn00561646_m1), NGF (Rn01533872_m1), nuclear factor kappa B p65 subunit (NFκB p65) (#Rn01502266_m1), nuclear factor erythroid-derived 2-like 2 (Nrf2) (#Rn00582415_m1), peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α; #Rn00580241_m1), p21 (#Rn 00589996_m1), sirtuin 1 (SIRT1; #Rn01428096_ m1), superoxide dismutase 1 (SOD1; #Rn00566938_m1), superoxide dismutase 2 (SOD2; #Rn00690588_g1), SRY-box transcription factor 10 (SOX10; #Rn00569909_m1), TNF-α (#Rn99999017_m1), tyrosine protein kinase receptor A (TrkA; #Rn00572130_m1), tyrosine protein kinase receptors B (TrkB; #Rn01441749_m1), and vascular endothelial growth factor (VEGF) (#Rn01511602_m1) were performed using the TaqMan real-time RT-PCR system with inventory primers and probes, as referred for each gene. Target mRNA levels were normalized to β-actin levels. Results were analyzed employing the 2−ΔΔCt method [40] and expressed relative to the control levels.
Statistical Analyses
Results are presented as mean ± standard deviation (S.D). The normal distribution was confirmed by Shapiro–Wilk test and then data were statistically analyzed using Student’s t-test. P values < 0.05 were considered significant. *Indicates differences between control and sulforaphane groups. All analyses were performed using GraphPad Prism 9.
Results
Anti-inflammatory Effects of Sulforaphane in Hypothalamic Astrocyte Cultures from Aged Rats
First, we tested the cellular viability (measured by MTT reduction) and membrane integrity (by PI incorporation) of three concentrations of sulforaphane (1, 5, and 10 μM) for 24 h in astrocyte cultures. Only 10 μM of sulforaphane decreased MTT reduction and increased PI incorporation (data not shown). Therefore, based on these data and a previous study of our group with sulforaphane in C6 astroglial cells [36], we choose the safely applicable concentration of 5 μM of sulforaphane to treat aged hypothalamic astrocyte cultures. In addition, 5 μM is a commonly used dose since higher doses were proven to be cytotoxic in certain experimental models [28, 36, 37, 41].
In line with this, as shown in the Fig. 1a–d, sulforaphane was able to decrease both the release and mRNA expression of the pro-inflammatory mediators TNF-ɑ (P < 0.001 and P = 0.01) and IL-1β (both P < 0.001), and the release of IL-6 (P < 0.001; Fig. 1e). In contrast, sulforaphane increased the release and mRNA expression of the anti-inflammatory cytokine IL-10 (P < 0.001 and P = 0.004, respectively; Fig. 1g and h).
The mRNA expression of COX-2 was also measured and sulforaphane was able to decrease it (P = 0.002; Fig. 1i). In addition, we observed that sulforaphane increased the mRNA expression of both adenosine receptors A1 and A2A (P = 0.009 and P = 0.002; Fig. 1j and k), while A2B was not affected (Fig. 1l).
Antioxidant-Related Effects of Sulforaphane in Astrocyte Cultures from Aged Rats
Our results showed that sulforaphane did not induce changes in the mRNA expression of SOD 1 and 2 (Fig. 2a and b). However, we found that sulforaphane was able to increase the mRNA expression of GCL and PGC-1α (both P < 0.001; Fig. 2c and e), and to decrease the mRNA expression of iNOS (P < 0.001; Fig. 2d).
Effects of Sulforaphane on mRNA of Proteins Related to Astrocyte Markers
Our results showed that sulforaphane had not effect on GFAP and GS mRNA expressions (Fig. 3a and b). On the other hand, sulforaphane increased ALDHL1 (P < 0.001; Fig. 3c), SOX10 (P = 0.002; Fig. 3d) and VEGF (P = 0.002; Fig. 3f) mRNA expressions and dowregulated AQP4 (P = 0.03; Fig. 3e).
Effects of Sulforaphane on Trophic Factors
In this study, we observed that sulforaphane was able to increase the mRNA expression of GDNF and NGF (P < 0.001; Fig. 4d and f), in addition to increase the release of these trophic factors and BDNF (P < 0.001 for all; Fig. 4a, c and e). Moreover, sulforaphane increased TrkA mRNA expression (P < 0.001; Fig. 4g), but did not change TrkB mRNA levels (Fig. 4h).
Potential Mechanisms Associated with Sulforaphane-Induced Glioprotection
Sulforaphane has a broad range of protective effects, which have been linked to several signaling pathways, including Nrf2, which activate cytoprotective genes [26, 42]. Furthermore, Nrf2 is known to act as an upstream signal for NFκB and HO-1 [43]. Here, we observed that sulforaphane increased the mRNA expressions of Nrf2 and HO-1 (both P < 0.001; Fig. 5a and b), while in the p65 NFκB expression (P < 0.001; Fig. 5c) it showed an opposite effect.
In addition, sulforaphane did not alter SIRT1 mRNA expression (Fig. 5d), but decreased AMPK and p21 mRNA expressions (P = 0.004 and P < 0.001, respectively; Fig. 5e and f).
Effects of Sulforaphane on Hypothalamic Tissue
We further evaluated the effects of sulforaphane in hypothalamic tissue, by using a cell suspension containing both neurons and glial cells. The cell suspension was incubated with 5 μM sulforaphane for 1 h to maintain complete viability of this hypothalamic preparation. Specific parameters were evaluated, and some differences were observed. Figure 6 displays that sulforaphane increased the mRNA expression of TNF-ɑ (P < 0.001; Fig. 6a), IL-6 (P < 0.001; Fig. 6c), IL-10 (P < 0.001; Fig. 6d), Nrf2 (P < 0.001; Fig. 6f), HO-1 (P < 0.001; Fig. 6g), SIRT1 (P = 0.02; Fig. 6j), GDNF (P < 0.001; Fig. 6o), NGF (P = 0.002; Fig. 6p) and TrkA (P = 0.002; Fig. 6q). In contrast, sulforaphane decreased TrkB mRNA expression (P < 0.001; Fig. 6r) and did not alter IL-1β (Fig. 6b), COX-2 (Fig. 6e), p65 NFκB (Fig. 6h), GFAP (Fig. 6k), ALDHL1 (Fig. 6l), SOX10 (Fig. 6m), BDNF (Fig. 6n) and iNOS (Fig. 6i).
Discussion
Aging is closely associated with progressive changes in the biology of cells, including astrocytes. These cells are critical to maintain brain homeostasis, and accumulating evidence suggests the role of hypothalamic astrocytes in the progression of aging due to its specific activities on metabolic and inflammatory functions [11, 44,45,46,47]. Therefore, protective strategies have been increasingly explored, and the identification of glioprotective molecules, such as sulforaphane, can prevent and/or avoid early events associated with aging [17]. In this regard, this study investigated the effects of sulforaphane on hypothalamic astrocyte cultures and on hypothalamic tissue from aged Wistar rats (24 months). Our findings indicate that sulforaphane has significant anti-inflammatory, antioxidant, and trophic functions, and can mediate glioprotection in the hypothalamus of aged rats.
Sulforaphane exhibits hormetic properties, with opposite effects depending on its concentration. At low doses, such as 5 μM, sulforaphane has shown cytoprotective effects and did not affect cellular viability and integrity at concentrations at least up to 10 μM [36, 41, 48, 49], in agreement with our data. Higher concentrations of sulforaphane (20 μM or more) can induce cytotoxicity through several mechanisms, including excessive ROS production, autophagy, and cell cycle arrest [31, 48, 50]. Concerning the time of incubation, the use of primary astrocyte cultures derived from 24-months old Wistar rats involves some limitations to perform evaluations in different time points, thus we choose 24 h. Although we could not rule out earlier and/or later evaluations in our study, this time of incubation is reasonable to assume changes in gene expression and release of cytokines and trophic factors and is particularly interesting for action of glioprotective molecules, as our group have demonstrated [51,52,53,54,55,56,57].
Particularly regarding aging, sulforaphane can modulate the expression of various biomarkers linked to inflammation, cellular senescence, and oxidative stress. Here, we observed that sulforaphane was able to decrease p21 expression, an important senescence marker that is associated with interruption of cell proliferation and acceleration of inflammatory process [58], suggesting a potential approach for treating age-related conditions [58,59,60]. The aging process is also characterized by an increase in the inflammatory responses, and in line with this, our previous research has demonstrated that hypothalamic astrocytes from mature animals exhibit a pro-inflammatory profile of cytokine and chemokine production [11]. Many pro-inflammatory cytokines are induced through the NFκB pathway, and this transcription factor was enhanced in an age-dependent way in our previous study [11]. On the contrary, the downregulation of NFκB has been associated with an improvement of inflammatory conditions, and sulforaphane was able to decrease not only NFκB expression but also TNF-ɑ, IL-1β and IL-6. We also verified that sulforaphane decreased the mRNA expression of COX-2, an enzyme involved in the synthesis of inflammatory mediators, which may be a transcriptional target of NFκB [61]. On the contrary, in another study, sulforaphane increased the secretion of several chemokines in astrocytes subjected to a stress-induced premature senescence [62]. Therefore, the effects of this compound specifically on chemokines production and release might be better characterized in future studies using astrocytes derived from aged rats. Our results also showed an increase in the mRNA expression of both A1 and A2A adenosine receptors, which are known to have anti-inflammatory effects in glial cells [63, 64]. These results reinforce the glioprotective role of sulforaphane on astroglial cells.
Sulforaphane is a well-recognized activator of Nrf2, which is able to increase the expression of cytoprotective enzymes and factors that may control the redox balance and inflammatory process, such as GCL, NFκB, HO-1, among others [43, 65]. An upregulation of SOD1 and SOD2 by sulforaphane could be also expected [66]. However, it is important to note that astrocytes present highly adaptive antioxidant mechanisms thus changes in gene expression may be fast and transient, returning to basal levels after 24 h. Particularly regarding SOD1 and SOD2, based on previous in vitro data from C6 astroglial cells and cardiomyocytes, it is reasonable to assume that their gene expression, protein content and activity are not direct targets of sulforaphane modulation, since no differences in this antioxidant enzyme were observed in the two cellular models after sulforaphane incubation for 1, 4 or 24 h [27, 36].
HO-1 is responsible for producing cellular responses against stressful conditions and has been related to the protective effects of another glioprotective molecule, resveratrol [44, 67]. We observed that sulforaphane increased the mRNA expression of Nrf2 and HO-1, while p65 NFκB showed an opposite effect. It is noteworthy that Nrf2 and HO-1 exert an upstream control of NFκB, negatively modulating its activation. Together, Nrf2 and NFκB are considered the major transcriptional factors to induce cellular responses, including metabolic responses. In line with this, AMPK signaling is also closely related to energy balance, in addition to regulate mitochondrial function, detoxification, and antioxidant defenses, promoting cellular homeostasis and, consequently, healthy aging [68]. Although AMPK activation has been related to sulforaphane effects in adipose and hepatic tissues [69, 70], we observed a downregulation of AMPK expression in hypothalamic astrocytes. Of note, there is a crosstalk between Nrf2 and AMPK signaling, in which the activation of Nrf2 may supress AMPK mRNA levels [69], which may explain the downregulation induced by sulforaphane. Nrf2 also regulates the transcription of the enzyme GCL, which participates in the synthesis of glutathione, a crucial antioxidant molecule that protects cells from oxidative stress and is primarily produced by astrocytes in the CNS [71]. PGC-1α can also control the expression of various enzymes involved in eliminating reactive oxygen species, as well as the induction of mitochondrial biogenesis through interaction with Nrf2 and SIRT1 pathways [72, 73]. Here, sulforaphane was able to upregulate GCL and PGC-1α.
The expression of iNOS can be triggered by different stimuli, including inflammation, cytokines, and oxidative stress [74, 75]. Our observations indicated that sulforaphane reduces the mRNA expression of this enzyme, which is downstream of Nrf2/HO-1, being a relevant target in pathological processes associated with inflammation and redox imbalance [76]. Therefore, sulforaphane was effective in our experimental model in the modulation of different signaling pathways that may prevent age-dependent functional alterations in astrocytes. In addition, modulation of endogenous cellular defense mechanisms represents an innovative approach to therapeutic intervention in diseases causing chronic tissue damage, such as in neurodegeneration due to accumulation of toxic products (including β-amyloid and ɑ-synuclein) and in the aging process. Considering that sulforaphane can also attenuate microglial activation [77,78,79], it can help in the development of neuroprotective/glioprotective therapeutic strategies through Nrf2 stimulation to restore redox equilibrium or to control neuroinflammation in CNS [80,81,82,83].
Besides regulating important functions associated with inflammatory response and redox homeostasis in cultured astrocytes, sulforaphane was able to modulate the expression of glial markers. Although sulforaphane did not change the expression of classical astrocytic markers, such as GFAP and GS, it modulated the mRNA levels of ALDH1L1, SOX10, AQP4, and VEGF. ALDH1L1 is an enzyme highly expressed in astrocytes and sulforaphane was able to increase its expression. This enzyme participates in the folate metabolim, influencing nucleotide biosynthesis and, consequently, the responses of neuron and glial cells, with potential impacts in regenerative process of the brain [71, 84]. In our study, we also observed that sulforaphane increased mRNA expression of SOX10. Although this transcription factor has been used as a marker of oligodendrocyte precursor cells, it is also involved in the development and maintenance of various cell types in the nervous system, including astrocytes [85, 86]. It is important to note that SOX10 perfoms many essential functions, including remodeling neural plasticity [87, 88], and our data reinforce the effects of sulforaphane against the deficits observed in brain aging.
The water channel AQP4, a protein expressed by astrocytes, is an important astrocytic functional parameter [89]. In addition to its role in maintaining water homeostasis, this protein has also been associated with neuroinflammation and neurodegeneration processes [90, 91]. Previous work from our group has shown that astrocytes derived from adult animal cultures express more AQP4 than those from neonatal animals, and furthermore, we also demonstrated that compounds such as resveratrol is capable of decreasing the expression of this protein under such condition [44, 92]. This data supports the result found in the present study, since sulforaphane downregulated AQP4 expression, possibly to maintain astrocytic homeostasis. In addition, VEGF plays a crucial role in regulating the formation and maintenance of the neurovascular unit, promoting the formation of new blood vessels in response to injury or disease, which can restore blood flow to damaged areas of the brain [93]. Moreover, astrocyte-derived VEGF may regulate inflammation and there is an interplay between VEGF and Nrf2 [94], both upregulated by sulforaphane.
The production and release of trophic factors by astrocytes is involved in their support functions, since these factors play critical roles during development and seem to mediate protective and/or reparative responses in mature cells. Trophic factors can act through high-affinity cell surface receptors TrkA and TrkB, supporting survival and plasticity of neural cells, and preventing cell death after injuries. In this study, we observed that sulforaphane increased the mRNA expression and the extracellular levels of GDNF and NGF, in addition to increase BDNF release. Besides that, sulforaphane increased TrkA mRNA expression [95]. It has been reported that the decrease in BDNF, GDNF, and NGF may be associated with the pathophysiology of neurodegeneration, while their increased levels may protect neural cells against toxic insults. Additionally, the expression of the trophic factors may be modulated by Nrf2/HO-1, which were also increased by sulforaphane [17].
Finally, we demonstrated that sulforaphane modulated gene expression in hypothalamic cells ex vivo in a similar way compared to cultured astrocytes, although a different profile of expression was observed for some genes, such as TNF-ɑ, IL-6 and TrkB. Of note, these differences may be related to the presence of other cell types, such as microglia and neurons, and to the shorter incubation time with sulforaphane. However, in general, this set of experiments demonstrated the ability of sulforaphane in modulating genes involved in important astroglial functions and suggests astrocyte responses with a focus on neuron-glia communication. These findings lead to the conclusion that sulforaphane presents glioprotective effects on hypothalamic tissue by modulating the expression of crucial genes related to inflammation, antioxidant defense, and neurotrophic factors in astrocytes, as well as signaling pathways that regulate these effects. Although further studies are needed to fully understand the mechanisms of sulforaphane in the CNS, our work indicate that this compound can contribute to the functional maintenance of aged hypothalamic astrocyte. In summary, our study highlights the potential benefits of sulforaphane in promoting glioprotection, reinforcing that sulforaphane may represent a promising experimental therapeutic intervention against aging-related brain dysfunctions and contributing to a healthier brain aging.
Data Availability
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
References
Coll AP, Yeo GS (2013) The hypothalamus and metabolism: integrating signals to control energy and glucose homeostasis. Curr Opin Pharmacol 13:970–976. https://doi.org/10.1016/j.coph.2013.09.010
Zhao Z-D, Yang WZ, Gao C et al (2017) A hypothalamic circuit that controls body temperature. Proc Natl Acad Sci USA 114:2042–2047. https://doi.org/10.1073/pnas.1616255114
Ono D, Yamanaka A (2017) Hypothalamic regulation of the sleep/wake cycle. Neurosci Res 118:74–81. https://doi.org/10.1016/j.neures.2017.03.013
Shalev D, Melamed P (2020) The role of the hypothalamus and pituitary epigenomes in central activation of the reproductive axis at puberty. Mol Cell Endocrinol 518:111031. https://doi.org/10.1016/j.mce.2020.111031
Trachtman JN (2010) Vision and the hypothalamus. Optom J Am Optom Assoc 81:100–115. https://doi.org/10.1016/j.optm.2009.07.016
Rizzi M, Gambini O, Marras CE (2021) Posterior hypothalamus as a target in the treatment of aggression: from lesioning to deep brain stimulation. Handbook of Clinical Neurology. Elsevier, Amsterdam, pp 95–106
Brüning JC, Fenselau H (2023) Integrative neurocircuits that control metabolism and food intake. Science 381:eab17398. https://doi.org/10.1126/science.abl7398
Maroso M, Stern P (2023) Small and mighty: the hypothalamus. Science 382:386–387. https://doi.org/10.1126/science.adl3437
Zhang G, Li J, Purkayastha S et al (2013) Hypothalamic programming of systemic ageing involving IKK-β, NF-κB and GnRH. Nature 497:211–216. https://doi.org/10.1038/nature12143
Kim K, Choe HK (2019) Role of hypothalamus in aging and its underlying cellular mechanisms. Mech Ageing Dev 177:74–79. https://doi.org/10.1016/j.mad.2018.04.008
Santos CL, Roppa PHA, Truccolo P et al (2018) Age-dependent neurochemical remodeling of hypothalamic astrocytes. Mol Neurobiol 55:5565–5579. https://doi.org/10.1007/s12035-017-0786-x
Behfar Q, Ramirez Zuniga A, Martino-Adami PV (2022) Aging, senescence, and dementia. J Prev Alz Dis. https://doi.org/10.14283/jpad.2022.42
Frago LM, Gómez-Romero A, Collado-Pérez R et al (2024) Synergism between hypothalamic astrocytes and neurons in metabolic control. Physiology 39:208–217. https://doi.org/10.1152/physiol.00009.2024
Pérez-Alvarez A, Araque A (2013) Astrocyte-neuron interaction at tripartite synapses. CDT 14:1220–1224. https://doi.org/10.2174/13894501113149990203
Perea G, Navarrete M, Araque A (2009) Tripartite synapses: astrocytes process and control synaptic information. Trends Neurosci 32:421–431. https://doi.org/10.1016/j.tins.2009.05.001
Benarroch EE (2016) Astrocyte signaling and synaptic homeostasis: I: membrane channels, transporters, and receptors in astrocytes. Neurology 87:324–330. https://doi.org/10.1212/WNL.0000000000002875
Quincozes-Santos A, Santos CL, de Souza Almeida RR et al (2021) Gliotoxicity and glioprotection: the dual role of glial cells. Mol Neurobiol 58:6577–6592. https://doi.org/10.1007/s12035-021-02574-9
Argente-Arizón P, Guerra-Cantera S, Garcia-Segura LM et al (2017) Glial cells and energy balance. J Mol Endocrinol 58:R59–R71. https://doi.org/10.1530/JME-16-0182
Teschemacher AG, Gourine AV, Kasparov S (2015) A role for astrocytes in sensing the brain microenvironment and neuro-metabolic integration. Neurochem Res 40:2386–2393. https://doi.org/10.1007/s11064-015-1562-9
Santos CL, Bobermin LD, Souza DO, Quincozes-Santos A (2018) Leptin stimulates the release of pro-inflammatory cytokines in hypothalamic astrocyte cultures from adult and aged rats. Metab Brain Dis 33:2059–2063. https://doi.org/10.1007/s11011-018-0311-6
Leloup C, Allard C, Carneiro L et al (2016) Glucose and hypothalamic astrocytes: more than a fueling role? Neuroscience 323:110–120. https://doi.org/10.1016/j.neuroscience.2015.06.007
Bobermin LD, Sesterheim P, Da Costa DS et al (2024) Simvastatin differentially modulates glial functions in cultured cortical and hypothalamic astrocytes derived from interferon α/β receptor knockout mice. Neurochem Res 49:732–743. https://doi.org/10.1007/s11064-023-04073-w
Bobermin LD, Da Costa DS, De Moraes ADM et al (2024) Effect of metformin in hypothalamic astrocytes from an immunocompromised mice model. Biochimie. https://doi.org/10.1016/j.biochi.2024.04.005
Bones A, Rossiter J (2006) The enzymic and chemically induced decomposition of glucosinolates. Phytochemistry 67:1053–1067. https://doi.org/10.1016/j.phytochem.2006.02.024
Baralić K, Živanović J, Marić Đ et al (2024) Sulforaphane—a compound with potential health benefits for disease prevention and treatment: insights from pharmacological and toxicological experimental studies. Antioxidants 13:147. https://doi.org/10.3390/antiox13020147
Bai Y, Wang X, Zhao S et al (2015) Sulforaphane protects against cardiovascular disease via Nrf2 activation. Oxid Med Cell Longev 2015:1–13. https://doi.org/10.1155/2015/407580
Corssac GB, Campos-Carraro C, Hickmann A et al (2018) Sulforaphane effects on oxidative stress parameters in culture of adult cardiomyocytes. Biomed Pharmacother 104:165–171. https://doi.org/10.1016/j.biopha.2018.05.031
Fernandes RO, De Castro AL, Bonetto JHP et al (2016) Sulforaphane effects on postinfarction cardiac remodeling in rats: modulation of redox-sensitive prosurvival and proapoptotic proteins. J Nutr Biochem 34:106–117. https://doi.org/10.1016/j.jnutbio.2016.05.004
Mann GE (2014) Nrf2-mediated redox signalling in vascular health and disease. Free Radical Biol Med 75:S1. https://doi.org/10.1016/j.freeradbiomed.2014.10.595
Poletto Bonetto JH, Luz De Castro A, Fernandes RO et al (2022) Sulforaphane effects on cardiac function and calcium-handling–related proteins in 2 experimental models of heart disease: ischemia-reperfusion and infarction. J Cardiovasc Pharmacol 79:325–334. https://doi.org/10.1097/FJC.0000000000001191
Asif Ali M, Khan N, Kaleem N et al (2023) Anticancer properties of sulforaphane: current insights at the molecular level. Front Oncol 13:1168321. https://doi.org/10.3389/fonc.2023.1168321
Hu R, Hebbar V, Kim B-R et al (2004) In vivo pharmacokinetics and regulation of gene expression profiles by isothiocyanate sulforaphane in the rat. J Pharmacol Exp Ther 310:263–271. https://doi.org/10.1124/jpet.103.064261
Tarozzi A, Angeloni C, Malaguti M et al (2013) Sulforaphane as a potential protective phytochemical against neurodegenerative diseases. Oxid Med Cell Longev 2013:1–10. https://doi.org/10.1155/2013/415078
Benedict AL, Mountney A, Hurtado A et al (2012) Neuroprotective effects of sulforaphane after contusive spinal cord injury. J Neurotrauma 29:2576–2586. https://doi.org/10.1089/neu.2012.2474
Carrasco-Pozo C, Tan KN, Borges K (2015) Sulforaphane is anticonvulsant and improves mitochondrial function. J Neurochem 135:932–942. https://doi.org/10.1111/jnc.13361
Bobermin LD, Weber FB, dos Santos TM et al (2022) Sulforaphane induces glioprotection after LPS challenge. Cell Mol Neurobiol 42:829–846. https://doi.org/10.1007/s10571-020-00981-5
Danilov CA, Chandrasekaran K, Racz J et al (2009) Sulforaphane protects astrocytes against oxidative stress and delayed death caused by oxygen and glucose deprivation. Glia 57:645–656. https://doi.org/10.1002/glia.20793
Verkerk R, Schreiner M, Krumbein A et al (2009) Glucosinolates in brassica vegetables: the influence of the food supply chain on intake, bioavailability and human health. Mol Nutr Food Res 53:S219–S219. https://doi.org/10.1002/mnfr.200800065
Bobermin LD, Quincozes-Santos A, Guerra MC et al (2012) Resveratrol prevents ammonia toxicity in astroglial cells. PLoS ONE 7:e52164. https://doi.org/10.1371/journal.pone.0052164
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Fernandes RO, Bonetto JHP, Baregzay B et al (2015) Modulation of apoptosis by sulforaphane is associated with PGC-1α stimulation and decreased oxidative stress in cardiac myoblasts. Mol Cell Biochem 401:61–70. https://doi.org/10.1007/s11010-014-2292-z
Liu J, Chandaka GK, Zhang R, Parfenova H (2020) Acute antioxidant and cytoprotective effects of sulforaphane in brain endothelial cells and astrocytes during inflammation and excitotoxicity. Pharmacol Res Perspect. https://doi.org/10.1002/prp2.630
Wakabayashi N, Slocum SL, Skoko JJ et al (2010) When NRF2 talks, who’s listening? Antioxid Redox Signal 13:1649–1663. https://doi.org/10.1089/ars.2010.3216
Sovrani V, Bobermin LD, Santos CL et al (2022) Effects of long-term resveratrol treatment in hypothalamic astrocyte cultures from aged rats. Mol Cell Biochem. https://doi.org/10.1007/s11010-022-04585-z
Palmer AL, Ousman SS (2018) Astrocytes and Aging. Front Aging Neurosci 10:337. https://doi.org/10.3389/fnagi.2018.00337
Santos CL, Bobermin LD, Quincozes-Santos A (2024) Aging changes the expression of adenosine receptors, insulin-like growth factor 1 (IGF1), and hypoxia-inducible factor 1α (HIF1α) in hypothalamic astrocyte cultures. Aging Brain 5:100104. https://doi.org/10.1016/j.nbas.2023.100104
Makarava N, Mychko O, Molesworth K et al (2023) Region-specific homeostatic identity of astrocytes is essential for defining their response to pathological insults. Cells 12:2172. https://doi.org/10.3390/cells12172172
Zanichelli F, Capasso S, Cipollaro M et al (2012) Dose-dependent effects of R-sulforaphane isothiocyanate on the biology of human mesenchymal stem cells, at dietary amounts, it promotes cell proliferation and reduces senescence and apoptosis, while at anti-cancer drug doses, it has a cytotoxic effect. Age 34:281–293. https://doi.org/10.1007/s11357-011-9231-7
Han Z, Xu Q, Li C, Zhao H (2017) Effects of sulforaphane on neural stem cell proliferation and differentiation. Genesis 55:e23022. https://doi.org/10.1002/dvg.23022
Sestili P, Fimognari C (2015) Cytotoxic and antitumor activity of sulforaphane: the role of reactive oxygen species. Biomed Res Int 2015:1–9. https://doi.org/10.1155/2015/402386
Bobermin LD, de Souza Almeida RR, Weber FB et al (2022) Lipopolysaccharide induces gliotoxicity in hippocampal astrocytes from aged rats: insights about the glioprotective roles of resveratrol. Mol Neurobiol 59:1419–1439. https://doi.org/10.1007/s12035-021-02664-8
Bellaver B, Bobermin LD, Souza DG et al (2016) Signaling mechanisms underlying the glioprotective effects of resveratrol against mitochondrial dysfunction. Biochimica et Biophysica Acta (BBA) Mol Basis Dis 1862:1827–1838. https://doi.org/10.1016/j.bbadis.2016.06.018
Arús BA, Souza DG, Bellaver B et al (2017) Resveratrol modulates GSH system in C6 astroglial cells through heme oxygenase 1 pathway. Mol Cell Biochem 428:67–77. https://doi.org/10.1007/s11010-016-2917-5
Quincozes-Santos A, Bobermin LD, Tonial RPL et al (2010) Effects of atypical (risperidone) and typical (haloperidol) antipsychotic agents on astroglial functions. Eur Arch Psychiatry Clin Neurosci 260:475–481. https://doi.org/10.1007/s00406-009-0095-0
Kleinkauf-Rocha J, Bobermin LD, Machado P de M et al (2013) Lipoic acid increases glutamate uptake, glutamine synthetase activity and glutathione content in C6 astrocyte cell line. Int j dev neurosci 31:165–170. https://doi.org/10.1016/j.ijdevneu.2012.12.006
Souza DG, Bellaver B, Bobermin LD et al (2016) Anti-aging effects of guanosine in glial cells. Purinergic Signal 12:697–706. https://doi.org/10.1007/s11302-016-9533-4
Bobermin LD, Sesterheim P, Da Costa DS et al (2023) Simvastatin differentially modulates glial functions in cultured cortical and hypothalamic astrocytes derived from interferon α/β receptor knockout mice. Neurochem Res. https://doi.org/10.1007/s11064-023-04073-w
Papismadov N, Gal H, Krizhanovsky V (2017) The anti-aging promise of p21. Cell Cycle 16:1997–1998. https://doi.org/10.1080/15384101.2017.1377500
Görg B, Karababa A, Shafigullina A et al (2015) Ammonia-induced senescence in cultured rat astrocytes and in human cerebral cortex in hepatic encephalopathy. Glia 63:37–50. https://doi.org/10.1002/glia.22731
Bellaver B, Souza DG, Souza DO, Quincozes-Santos A (2017) Hippocampal astrocyte cultures from adult and aged rats reproduce changes in glial functionality observed in the aging brain. Mol Neurobiol 54:2969–2985. https://doi.org/10.1007/s12035-016-9880-8
Poligone B, Baldwin AS (2001) Positive and negative regulation of NF-κB by COX-2. J Biol Chem 276:38658–38664. https://doi.org/10.1074/jbc.M106599200
Maciel-Barón LÁ, Morales-Rosales SL, Silva-Palacios A et al (2018) The secretory phenotype of senescent astrocytes isolated from Wistar newborn rats changes with anti-inflammatory drugs, but does not have a short-term effect on neuronal mitochondrial potential. Biogerontology 19:415–433. https://doi.org/10.1007/s10522-018-9767-3
Németh ZH, Lutz CS, Csóka B et al (2005) Adenosine augments IL-10 production by macrophages through an A2B receptor-mediated posttranscriptional mechanism. J Immunol 175:8260–8270. https://doi.org/10.4049/jimmunol.175.12.8260
Pasquini S, Contri C, Borea PA et al (2021) Adenosine and inflammation: here there and everywhere. IJMS 22:7685. https://doi.org/10.3390/ijms22147685
Niture SK, Khatri R, Jaiswal AK (2014) Regulation of Nrf2—an update. Free Radical Biol Med 66:36–44. https://doi.org/10.1016/j.freeradbiomed.2013.02.008
Bellezza I, Giambanco I, Minelli A, Donato R (2018) Nrf2-Keap1 signaling in oxidative and reductive stress. Biochimica et Biophysica Acta (BBA) Mol Cell Res 1865:721–733. https://doi.org/10.1016/j.bbamcr.2018.02.010
Quincozes-Santos A, Bobermin LD, Latini A et al (2013) Resveratrol protects C6 astrocyte cell line against hydrogen peroxide-induced oxidative stress through heme oxygenase 1. PLoS ONE 8:e64372. https://doi.org/10.1371/journal.pone.0064372
Petsouki E, Cabrera SNS, Heiss EH (2022) AMPK and NRF2: Interactive players in the same team for cellular homeostasis? Free Radical Biol Med 190:75–93. https://doi.org/10.1016/j.freeradbiomed.2022.07.014
Masuda M, Yoshida-Shimizu R, Mori Y et al (2022) Sulforaphane induces lipophagy through the activation of AMPK-mTOR-ULK1 pathway signaling in adipocytes. J Nutr Biochem 106:109017. https://doi.org/10.1016/j.jnutbio.2022.109017
Zhang Y, Wu Q, Liu J et al (2022) Sulforaphane alleviates high fat diet-induced insulin resistance via AMPK/Nrf2/GPx4 axis. Biomed Pharmacother 152:113273. https://doi.org/10.1016/j.biopha.2022.113273
Dahal A, Govindarajan K, Kar S (2023) Administration of kainic acid differentially alters astrocyte markers and transiently enhanced phospho-tau level in adult rat hippocampus. Neuroscience 516:27–41. https://doi.org/10.1016/j.neuroscience.2023.02.010
Scarpulla RC (2011) Metabolic control of mitochondrial biogenesis through the PGC-1 family regulatory network. Biochimica et Biophysica Acta (BBA) Mol Cell Res 1813:1269–1278. https://doi.org/10.1016/j.bbamcr.2010.09.019
Aguirre-Rueda D, Guerra-Ojeda S, Aldasoro M et al (2015) Astrocytes protect neurons from Aβ 1–42 peptide-induced neurotoxicity increasing TFAM and PGC-1 and decreasing PPAR-γ and SIRT-1. Int J Med Sci 12:48–56. https://doi.org/10.7150/ijms.10035
Calabrese V, Cornelius C, Dinkova-Kostova AT et al (2010) Cellular stress responses, the hormesis paradigm, and vitagenes: novel targets for therapeutic intervention in neurodegenerative disorders. Antioxid Redox Signal 13:1763–1811. https://doi.org/10.1089/ars.2009.3074
Calabrese V, Mancuso C, Calvani M et al (2007) Nitric oxide in the central nervous system: neuroprotection versus neurotoxicity. Nat Rev Neurosci 8:766–775. https://doi.org/10.1038/nrn2214
Calabrese V, Giordano J, Signorile A et al (2016) Major pathogenic mechanisms in vascular dementia: roles of cellular stress response and hormesis in neuroprotection. J Neurosci Res 94:1588–1603. https://doi.org/10.1002/jnr.23925
Tufekci KU, Ercan I, Isci KB et al (2021) Sulforaphane inhibits NLRP3 inflammasome activation in microglia through Nrf2-mediated miRNA alteration. Immunol Lett 233:20–30. https://doi.org/10.1016/j.imlet.2021.03.004
Yang Y, Zhang J, Yang C et al (2023) Sulforaphane attenuates microglia-mediated neuronal damage by down-regulating the ROS/autophagy/NLRP3 signal axis in fibrillar Aβ-activated microglia. Brain Res 1801:148206. https://doi.org/10.1016/j.brainres.2022.148206
Qin S, Yang C, Huang W et al (2018) Sulforaphane attenuates microglia-mediated neuronal necroptosis through down-regulation of MAPK/NF-κB signaling pathways in LPS-activated BV-2 microglia. Pharmacol Res 133:218–235. https://doi.org/10.1016/j.phrs.2018.01.014
Concetta Scuto M, Mancuso C, Tomasello B et al (2019) Curcumin, hormesis and the nervous system. Nutrients 11:2417. https://doi.org/10.3390/nu11102417
Mancuso C, Capone C, Ranieri SC et al (2008) Bilirubin as an endogenous modulator of neurotrophin redox signaling. J of Neuroscience Research 86:2235–2249. https://doi.org/10.1002/jnr.21665
Butterfield DA, Boyd-Kimball D, Reed TT (2023) Cellular stress response (hormesis) in response to bioactive nutraceuticals with relevance to Alzheimer disease. Antioxid Redox Signal. https://doi.org/10.1089/ars.2022.0214
Calabrese EJ, Kozumbo WJ (2021) The phytoprotective agent sulforaphane prevents inflammatory degenerative diseases and age-related pathologies via Nrf2-mediated hormesis. Pharmacol Res 163:105283. https://doi.org/10.1016/j.phrs.2020.105283
Yang Y, Vidensky S, Jin L et al (2011) Molecular comparison of GLT1+ and ALDH1L1+ astrocytes in vivo in astroglial reporter mice. Glia 59:200–207. https://doi.org/10.1002/glia.21089
Bhattarai C, Poudel PP, Ghosh A, Kalthur SG (2022) Comparative role of SOX10 gene in the gliogenesis of central, peripheral, and enteric nervous systems. Differentiation 128:13–25. https://doi.org/10.1016/j.diff.2022.09.001
Reiprich S, Wegner M (2015) From CNS stem cells to neurons and glia: Sox for everyone. Cell Tissue Res 359:111–124. https://doi.org/10.1007/s00441-014-1909-6
Souza DG, Bellaver B, Terra SR et al (2017) In vitro adult astrocytes are derived from mature cells and reproduce in vivo redox profile. J of Cellular Biochemistry 118:3111–3118. https://doi.org/10.1002/jcb.26028
Wasinski F, Tavares MR, Gusmao DO et al (2023) Central growth hormone action regulates neuroglial and proinflammatory markers in the hypothalamus of male mice. Neurosci Lett 806:137236. https://doi.org/10.1016/j.neulet.2023.137236
Salman MM, Kitchen P, Halsey A et al (2022) Emerging roles for dynamic aquaporin-4 subcellular relocalization in CNS water homeostasis. Brain 145:64–75. https://doi.org/10.1093/brain/awab311
Fukuda AM, Badaut J (2012) Aquaporin 4: a player in cerebral edema and neuroinflammation. J Neuroinflammation 9:771. https://doi.org/10.1186/1742-2094-9-279
Yang J, Zhang R, Shi C et al (2017) AQP4 association with amyloid deposition and astrocyte pathology in the Tg-ArcSwe mouse model of Alzheimer’s disease. JAD 57:157–169. https://doi.org/10.3233/JAD-160957
Bobermin LD, Roppa RHA, Gonçalves C-A, Quincozes-Santos A (2020) Ammonia-induced glial-inflammaging. Mol Neurobiol 57:3552–3567. https://doi.org/10.1007/s12035-020-01985-4
Wuestefeld R, Chen J, Meller K et al (2012) Impact of vegf on astrocytes: analysis of gap junctional intercellular communication, proliferation, and motility. Glia 60:936–947. https://doi.org/10.1002/glia.22325
Li L, Pan H, Wang H et al (2016) Interplay between VEGF and Nrf2 regulates angiogenesis due to intracranial venous hypertension. Sci Rep 6:37338. https://doi.org/10.1038/srep37338
Lanfranconi S, Locatelli F, Corti S et al (2011) Growth factors in ischemic stroke. J Cell Mol Med 15:1645–1687. https://doi.org/10.1111/j.1582-4934.2009.00987.x
Acknowledgements
The authors of this study are supported by Universidade Federal do Rio Grande do Sul (UFRGS), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS), and Instituto Nacional de Ciência e Tecnologia para Excitotoxicidade e Neuroproteção (INCTEN/CNPq).
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This research was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS) and Instituto Nacional de Ciência e Tecnologia para Excitotoxicidade e Neuroproteção.
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CLS, ABK, LDB, and AQS conceptualized the study. CLS, FBW, and LDB performed the experiments. CLS, LDB, and AQS performed statistical analysis and written the original draft of the manuscript. ABK and AQS provided resources and materials/chemicals. All authors revised, edited, and approved the manuscript.
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Santos, C.L., Weber, F.B., Belló-Klein, A. et al. Glioprotective Effects of Sulforaphane in Hypothalamus: Focus on Aging Brain. Neurochem Res 49, 2505–2518 (2024). https://doi.org/10.1007/s11064-024-04196-8
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DOI: https://doi.org/10.1007/s11064-024-04196-8