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SUMMARY: Hypoxic preconditioning is known to induce neuroprotection, but its effects and pathways in chronic brain pathology still unknown. The aim was to establish an involvement of a7 subunit of nicotinic acetylcholine receptors (a7nAchRs), and sirtuins of 1 (SIRT1) and 3 (SIRT3) types in the effects of hypoxic hypobaric preconditioning on brain damage in mice with chronic cerebral hypoperfusion caused by the left common carotid artery occlusion. The male C57/6j (C57, wild type) and a7nAchRs(-/-) mice were divided to six experimental groups (10 mice per group): sham-operated C57, C57 with chronic cerebral hypoperfusion, C57 with hypoxic hypobaric preconditioning and chronic cerebral hypoperfusion, sham-operated a7nAchRs(-/-) mice, a7nAchRs(-/-) with chronic cerebral hypoperfusion, a7nAchRs(-/-) with hypoxic hypobaric preconditioning and chronic cerebral hypoperfusion. For preconditioning, mice were exposed to hypoxia by "lifting" in barochamber to simulated altitude of 5600 m a.s.l. for 1 h/day on 3 consecutive days before surgical manipulation. Expressions of SIRT1, SIRT3 in brain tissue, and histopathological changes of the hippocampi were examined. It was shown that 8-week chronic hypoperfusion of the brain, caused by unilateral occlusion of the common carotid artery, was accompanied by injury to the neurons of the hippocampi of both hemispheres, which was more pronounced on the side of the occlusion. This damage, as well as the mechanisms of neuroprotection induced by hypoxic preconditioning, were maintained for at least 8 weeks by mechanisms mediated through a7nAChRs. Deficite of a7nAChRs was accompanied with reduction of neuronal damage caused CCH in 8 weeks, as well as preconditioning effects, and lead to compensatory activation of regulatory and protective mechanisms mediated by SIRT1, in normal conditions and in CCH. In wild-type (C57) mice, protective mechanisms in CCH were realized to a greater extent by increased expression of SIRT3 in both hemispheres of the brain.
Se sabe que el precondicionamiento hipóxico induce neuroprotección, pero aún se desconocen sus efectos y vías en la patología cerebral crónica. El objetivo fue establecer la participación de la subunidad a7 de los receptores nicotínicos de acetilcolina (a7nAchR) y las sirtuinas de tipo 1 (SIRT1) y 3 (SIRT3) en los efectos del precondicionamiento hipóxico hipobárico sobre el daño cerebral en ratones con hipoperfusión cerebral crónica causada por la oclusión de la arteria carótida común izquierda. Los ratones macho C57/6j (C57, tipo salvaje) y a7nAchRs(-/-) se dividieron en seis grupos experimentales (10 ratones por grupo): C57 con operación simulada, C57 con hipoperfusión cerebral crónica, C57 con precondicionamiento hipobárico hipóxico y crónica. hipoperfusión cerebral, ratones a7nAchRs(-/-) operados de forma simulada, a7nAchRs(-/-) con hipoperfusión cerebral crónica, a7nAchRs(-/-) con precondicionamiento hipobárico hipóxico e hipoperfusión cerebral crónica. Para el preacondicionamiento, los ratones fueron expuestos a hipoxia "levantándolos" en una cámara de barro a una altitud simulada de 5600 m s.n.m. durante 1 h/día durante 3 días consecutivos antes de la manipulación quirúrgica. Se examinaron las expresiones de SIRT1, SIRT3 en tejido cerebral y los cambios histopatológicos de los hipocampos. Se demostró que la hipoperfusión cerebral crónica de 8 semanas, causada por la oclusión unilateral de la arteria carótida común, se acompañaba de lesión de las neuronas del hipocampo de ambos hemisferios y que era más pronunciada en el lado de la oclusión. Este daño, así como los mecanismos de neuroprotección inducidos por el precondicionamiento hipóxico, se mantuvieron durante al menos 8 semanas mediante mecanismos mediados por a7nAChR. El déficit de a7nAChR se acompañó de una reducción del daño neuronal causado por CCH en 8 semanas, así como de efectos de precondicionamiento, y condujo a una activación compensatoria de mecanismos reguladores y protectores mediados por SIRT1, en condiciones normales y en CCH. En ratones de tipo salvaje (C57), los mecanismos de protección en CCH se realizaron en mayor medida mediante una mayor expresión de SIRT3 en ambos hemisfe- rios del cerebro.
Subject(s)
Animals , Mice , Brain Ischemia , Sirtuin 1/metabolism , Sirtuin 3/metabolism , alpha7 Nicotinic Acetylcholine Receptor/metabolism , Hypoxia , Cerebrovascular Circulation , Blotting, Western , Carotid StenosisABSTRACT
SUMMARY: Senile osteoporosis is mainly caused by reduced osteoblast differentiation and has become the leading cause of fractures in the elderly worldwide. Natural organics are emerging as a potential option for the prevention and treatment of osteoporosis. This study was designed to study the effect of resveratrol on osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in osteoporosis mice. A mouse model of osteoporosis was established by subcutaneous injection of dexamethasone and treated with resveratrol administered by gavage. In vivo and in vitro, we used western blot to detect protein expression, and evaluated osteogenic differentiation of BMSCs by detecting the expression of osteogenic differentiation related proteins, calcium deposition, ALP activity and osteocalcin content. Resveratrol treatment significantly increased the body weight of mice, the level of serum Ca2+, 25(OH)D and osteocalcin, ration of bone weight, bone volume/total volume, trabecular thickness, trabecular number, trabecular spacing and cortical thickness in osteoporosis mice. In BMSCs of osteoporosis mice, resveratrol treatment significantly increased the expression of Runx2, osterix (OSX) and osteocalcin (OCN) protein, the level of calcium deposition, ALP activity and osteocalcin content. In addition, resveratrol treatment also significantly increased the expression of SIRT1, p-PI3K / PI3K and p-AKT / AKT in BMSCs of osteoporosis mice. In vitro, resveratrol increased the expression of SIRT1, p-PI3K / PI3K and p-AKT / AKT, Runx2, OSX and OCN protein, the level of calcium deposition, ALP activity and osteocalcin content in BMSCs in a concentration-dependent manner, while SIRT1 knockdown significantly reversed the effect of resveratrol. Resveratrol can attenuate osteoporosis by promoting osteogenic differentiation of bone marrow mesenchymal stem cells, and the mechanism may be related to the regulation of SIRT1/PI3K/AKT pathway.
La osteoporosis senil es causada principalmente por una diferenciación reducida de osteoblastos y se ha convertido en la principal causa de fracturas en las personas mayores en todo el mundo. Los productos orgánicos naturales están surgiendo como una opción potencial para la prevención y el tratamiento de la osteoporosis. Este estudio fue diseñado para estudiar el efecto del resveratrol en la diferenciación osteogénica de las células madre mesenquimales de la médula ósea (BMSC) en ratones con osteoporosis. Se estableció un modelo de osteoporosis en ratones mediante inyección subcutánea de dexametasona y se trató con resveratrol administrado por sonda. In vivo e in vitro, utilizamos Western blot para detectar la expresión de proteínas y evaluamos la diferenciación osteogénica de BMSC detectando la expresión de proteínas relacionadas con la diferenciación osteogénica, la deposición de calcio, la actividad de ALP y el contenido de osteocalcina. El tratamiento con resveratrol aumentó significativamente el peso corporal de los ratones, el nivel sérico de Ca2+, 25(OH)D y osteocalcina, la proporción de peso óseo, el volumen óseo/ volumen total, el espesor trabecular, el número trabecular, el espaciado trabecular y el espesor cortical en ratones con osteoporosis. En BMSC de ratones con osteoporosis, el tratamiento con resveratrol aumentó significativamente la expresión de las proteínas Runx2, osterix (OSX) y osteocalcina (OCN), el nivel de deposición de calcio, la actividad de ALP y el contenido de osteocalcina. Además, el tratamiento con resveratrol también aumentó significativamente la expresión de SIRT1, p-PI3K/PI3K y p-AKT/AKT en BMSC de ratones con osteoporosis. In vitro, el resveratrol aumentó la expresión de las proteínas SIRT1, p-PI3K/PI3K y p- AKT/AKT, Runx2, OSX y OCN, el nivel de deposición de calcio, la actividad de ALP y el contenido de osteocalcina en BMSC de manera dependiente de la concentración, mientras que La caída de SIRT1 revirtió significativamente el efecto del resveratrol. El resveratrol puede atenuar la osteoporosis al promover la diferenciación osteogénica de las células madre mesenquimales de la médula ósea, y el mecanismo puede estar relacionado con la regulación de la vía SIRT1/PI3K/AKT.
Subject(s)
Animals , Male , Mice , Osteoporosis/drug therapy , Resveratrol/administration & dosage , Osteogenesis/drug effects , Cell Differentiation/drug effects , Blotting, Western , Disease Models, Animal , Sirtuin 1 , Mesenchymal Stem Cells/cytology , Mesenchymal Stem Cells/drug effects , Resveratrol/pharmacology , Mice, Inbred C57BLABSTRACT
ObjectiveThe lung mesenchymal stem cells (LMSCs) induced by D-galactose (D-gal) were intervened by Wenfei Huaxian decoction-containing serum to explore the mechanism of Wenfei Huaxian decoction in delaying the senescence of LMSCs through the nicotinamide phosphoribosyltransferase/silent information regulator 1 (NAMPT/SIRT1) signaling pathway. MethodWenfei Huaxian decoction-containing serum was prepared. LMSCs were isolated by gradient density centrifugation, and they were cultured and identified in vitro. The senescence model in vitro was established by stimulating cells via D-gal for 24 h. LMSCs cells were modeled after being treated with different volume fractions (5%, 10%, 20%, 40%, and 80%) of Wenfei Huaxian decoction-containing serum for 24 h, and the cell proliferation level was detected by methyl thiazolyl tetrazolium (MTT) method. The cells were randomly divided into blank serum group, model group, and high, medium, and low dose groups of Wenfei Huaxian decoction-containing serum. Senescence-associated β-galactosidase (SA-β-gal) staining was used to detect the senescence of LMSCs in each group. The content of NAD + was detected by colorimetry. The levels of senescence-associated factors (p16 and p53), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) in cell culture supernatant were detected by enzyme-linked immunosorbent assay (ELISA). Western blot was used to detect the relative expression of senescence-associated proteins and NAMPT/SIRT1 signaling pathway-related proteins. ResultCompared with the blank serum group, the proliferation of LMSCs was significantly inhibited after D-gal stimulation for 24 h (P<0.01). Compared with the model group, the proliferation of LMSCs could be promoted after intervention with the corresponding Wenfei Huaxian decoction-containing serum (P<0.05, P<0.01). Compared with the blank serum group, the SA-β-gal staining of LMSCs in the model group after D-gal stimulation was enhanced, and the content of NAD+ was increased (P<0.01). The expression levels of senescence factors p16 and p53, as well as SASP pro-inflammatory factors IL-6 and TNF-α in the cell culture supernatant, were significantly increased (P<0.01). The expression of senescence-associated proteins p16, p21, and p53 increased (P<0.01), and the protein expression of NAMPT, SIRT1, peroxisome proliferator-activated receptor γ coactivator 1α (PGC-1α), and forkhead box family transcription factor O1 (FoxO1) decreased (P<0.01). Compared with the model group, the SA-β-gal staining of LMSCs in each group of Wenfei Huaxian decoction-containing serum was significantly reduced, and the content of NAD+ was decreased (P<0.01). The senescence factors (p16 and p53) and inflammatory factors (IL-6 and TNF-α) in the cell culture supernatant were significantly decreased (P<0.01). The expression of senescence-associated proteins (P16, P21, and P53) decreased (P<0.05, P<0.01). The protein expressions of NAMPT, SIRT1, PGC-1α, and FoxO1 were significantly up-regulated (P<0.05, P<0.01). ConclusionWenfei Huaxian decoction can alleviate senescence and inflammatory response damage of D-gal-induced LMSCs, and its mechanism may be related to the regulation of the NAMPT/SIRT1 signaling pathway.
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Objective To investigate the role and regulatory mechanism of stress-inducing protein 1(SESN1)in liver gluconeogenesis of fasting mice.Methods RT-qPCR was used to detect mRNA expression of SESN1 in liver tissues of C57BL/6J mice and primary mouse hepatocytes treated with forskolin(Fsk)and dexamethasone(Dex).HepG2 cells were transfected with plasmids and the effects of SESN1 overexpression on mRNA expression of gluconeogenesis related genes PGC-1α,PEPCK and G6Pase was detected by RT-qPCR.The effect of SESN1 on the promoter activity of PGC-1α in HepG2 cells was studied using a dual luciferase reporter system.The effect of SESN1 on PGC-1α deacetylation was detected by overexpression of SESN1 and inhibition of SIRT1 expression.By knocking down SIRT1 expression,we detected whether it mediated the changes in mRNA levels of SESN1 in-duced gluconeogenesis related genes.Results The mRNA expression of SESN1 was significantly increased in liver tissues of starved C57BL/6J mice and in primary hepatocytes treated with Fsk and Dex(P<0.001).Over-expression of SESN1 in HepG2 cells promoted mRNA expression of PGC-1α,PEPCK and G6Pase(P<0.001)and promoter activity of PGC-1α(P<0.001).Over-expression of SESN1 decreased the acetylation level of PGC-1α in primary hepatocytes.Sirt family inhibitors NAM and shRNA adenovirus interfered with SIRT1 expression respective-ly,and antagonized the deacetylation effect of SESN1 on PGC-1α.The expression of PGC-1α,PEPCK and G6Pase induced by SIRT1 was also significantly impaired(P<0.000 1).Conclusions SESN1 regulates liver gluconeogene-sis in mice with a SIRT1-dependent mechanism.
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Objective To investigate the effects of puerarin on myocardial ischemia-reperfusion injury and its mecha-nism.Methods Molecular docking and dynamics simulation were utilized to predict the binding potential of puerarin and SIRT1.A myocardial ischemia-reperfusion model was established in SD rats by ligating the anterior descending branch of the left coronary artery.The protective effect of puerarin on myocardial injury was observed,and the therapeutic effect of puerarin was compared after inhibition of SIRT1 expression.The infarct volume was detected using 2,3,5-triphenyltetrazolium chloride(TTC)staining.The apoptosis rate and SIRT1 expression of cardiomyocytes were detected by using TUNEL combined with im-munofluorescence.Transmission electron microscope was used to observe the myocardial ultrastructure.Western blot was per-formed to detect the expression of ferroptosis-related proteins.Results Molecular docking studies confirmed the formation of stable complexes between puerarin and SIRT1.Puerarin treatment significantly increased myocardial ischemia-reperfusion injury through upregulation of SIRT1,SLC7A11 and GPX4 expression,and downregulation of IREB2 expression in rats.The protec-tive effect of puerarin on myocardium was abolished once SIRT1 protein expression was inhibited.Conclusion Molecular doc-king and molecular dynamics simulation techniques can accurately predict the interaction of puerarin,and the main target SIRT1.Puerarin inhibits ferroptosis by activating SIRT1 pathway,thereby alleviating myocardial ischemia-reperfusion injury.
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Objective To observe the effect of sirtuin 1(SIRT1)on rat myocardial fibrosis induced by pressure overload and the proliferation of cardiac fibroblasts induced by angiotensin Ⅱ(Ang Ⅱ),and to explore the molecular mechanisms.Methods The pressure overload-induced myocardial fibrosis was established by abdominal aorta constriction(AAC)procedure in vi-vo.After treatment with SIRT1 activator,the myocardial interstitial fibrosis and the collagen volume fraction were evaluated by Masson's trichrome staining.The protein expressions of TGF-β1/Smads were determined by immunohistochemical analy-sis.After in vitro intervention of Ang Ⅱ or Ang Ⅱ with SIRT1 activator,the fibroblasts proliferation was detected by MTT as-say.The mRNA and protein expressions of collagen Ⅰ/Ⅲ(Col1α1/3α1),SIRT1 and TGF-β1/Smads in myocardial tissue and fi-broblasts were evaluated by qRT-PCR and Western blotting.Results Compared with the sham operation group,myocardial in-terstitial fibrosis was significantly observed in the pressure overload model group,myocardial collagen volume fraction was in-creased,expressions of Col1α1/3α1 and TGF-β1/Smads were significantly increased,and SIRT1 expression was decreased.After the intervention of SRT1720,SIRT1 activator could improve the myocardial interstitial fibrosis induced by pressure overload,downregulate the expressions of Col1α1/3α1 and TGF-β1/Smads,and upregulate the expression of SIRT1.Meanwhile,correla-tion analysis showed that the protein expression of SIRT1 was negatively correlated with the expression of TGF-β1.In addition,SRT1720 also inhibited Ang Ⅱ-induced fibroblast proliferation and increased expression of Col1α1/3α1 and TGF-β1.Conclusion Activation of SIRT1 inhibits pressure overload-induced myocardial fibrosis and Ang Ⅱ-induced fibroblasts proliferation via regu-lation of the TGF-β1/Smads signaling pathway.
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BACKGROUND:Pretreatment with moxibustion is a preventive treatment in traditional Chinese medicine.Pretreatment with moxibustion at the onset of prodromal symptoms can significantly reduce the symptoms and delay the onset of many diseases,but the exact mechanism remains to be studied. OBJECTIVE:To investigate the mechanism of SIRT1/FoxO3 pathway in moxibustion pretreatment to ameliorate oxidative stress injury in cerebral ischemia-reperfusion model rats. METHODS:Forty-eight Sprague-Dawley rats were randomly divided into sham-operated group,model group,moxibustion pretreatment group,and moxibustion pretreatment+EX527(SIRT1 inhibitor)group,with 12 rats in each group.The moxibustion pretreatment group was given moxibustion with seed-sized moxa cone at Baihui,Dazhui,and Zusanli before modeling,three moxa-cones per acupoint,once a day for 7 days.In the model group,moxibustion pretreatment group and moxibustion pretreatment+EX527 group,the rat model of middle cerebral artery occlusion was made by suturing of the middle cerebral artery 30 minutes after the last moxibustion.After 2 hours of cerebral ischemia,the middle artery suture was removed and the rats were reperfused for 12 hours.In the sham-operated group,only the common carotid artery,internal carotid artery,and external carotid artery were dissected without suturing the middle cerebral artery.In the moxibustion pretreatment+EX527 group,EX527(15 mg/kg)was given intraperitoneally 30 minutes before each moxibustion.After 12 hours of reperfusion,the rats were scored for neurological deficits,and the cerebral infarct volume was calculated by 2,3,5-triphenyltetrazolium chloride staining method.The levels of oxidative stress factors in the infarcted tissues were detected by the kit method,and western-blot method was used to detect the expression levels of SIRT1,FoxO3,p-FoxO3 and brain-derived neurotrophic factor in the ischemic area of the cerebral cortex. RESULTS AND CONCLUSION:After 12 hours of reperfusion,the neurobehavioral score in the model group was significantly higher than that in the sham-operated group(P<0.01),while the score in the moxibustion pretreatment group was significantly lower than that in the model group(P<0.01)and moxibustion pretreatment+EX527 group(P<0.05).There were no obvious infarct foci in the brain tissue of the sham-operated rats,but obvious ischemic foci were observed in the right side of the brain tissue of the rats in the model group(P<0.01).The right infarct volume in the moxibustion pretreatment group was significantly reduced compared with the model group(P<0.01),while the right infarct volume in the moxibustion pretreatment+EX527 group was significantly enlarged compared with the moxibustion pretreatment group.After 12 hours of reperfusion,the level of malondialdehyde was significantly elevated(P<0.01)and the expression of superoxide dismutase was significantly decreased(P<0.01)in the model group compared with the sham-operated group.The levels of malondialdehyde was significantly decreased(P<0.01,P<0.05)and the expression of superoxide dismutase was significantly increased(P<0.01,P<0.05)in the moxibustion pretreatment group compared with the model group and the moxibustion pretreatment+EX527 group.Western blot results showed that the expression levels of SIRT1,FoxO3,p-FoxO3,and brain-derived neurotrophic factor proteins were significantly higher in the model group compared with the sham-operated group(P<0.01);compared with the model group,the expression levels of SIRT1,FoxO3,and brain-derived neurotrophic factor were significantly higher in the moxibustion pretreatment group(P<0.01),and p-FoxO3 expression was significantly lower(P<0.01);compared with the moxibustion pretreatment+EX527 group,the expression levels of SIRT1,FoxO3,and brain-derived neurotrophic factor were elevated in the moxibustion pretreatment group(P<0.05),and no statistically significant difference was found in the p-FoxO3 expression(P>0.05).To conclude,moxibustion pretreatment can significantly improve neurological function in rats after cerebral ischemia-reperfusion,and the mechanism may be related to the activation of SIRT1/FoxO3 pathway to reduce oxidative stress injury in the rat model of cerebral ischemia-reperfusion.
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BACKGROUND:Previous studies have shown that silent mating-type information regulator 2 homolog 1(SIRT1)-mechanistic target of rapamycin(mTOR)signaling plays an important role in the progression of osteoarthritis.Emodin has a protective effect on osteoarthritic chondrocytes. OBJECTIVE:To investigate the effects of emodin on the proliferation,apoptosis and oxidative stress of osteoarthritic chondrocytes based on the SIRT1-mTOR signaling. METHODS:Rat chondrocytes were isolated and cultured in vitro.Osteoarthritic chondrocyte model in vitro was induced by 10 ng/mL interleukin-1β.Cell counting kit-8 method was used to determine the viability of rat chondrocytes treated with 0,20,40,80,120,160 μmol/L emodin,and the appropriate concentration of emodin was selected.Rat chondrocytes isolated and cultured in vitro were randomly divided into control group,model group,low-dose emodin group,high-dose emodin group,EX527 group,and high-dose emodin+EX527 group.In vitro osteoarthritis model was constructed by induction of 10 ng/mL interleukin 1β in all groups except the control group.The cells in the latter four groups were correspondingly treated with emodin or/and EX527.The proliferation and apoptosis of chondrocytes in each group were detected by cell counting kit-8,Edu staining and flow cytometry respectively.The relative content of reactive oxygen species and the levels of malondialdehyde,superoxide dismutase,catalase,and glutathione peroxidase in chondrocytes of rats in each group were measured with the kit.The expression of proteins related to cell matrix degradation,apoptosis and the SIRT1-mTOR pathway-related proteins in each group were detected by western blot. RESULTS AND CONCLUSION:Compared with the control group,the survival rate of chondrocytes,the positive rate of Edu,the levels of superoxide dismutase,catalase,and glutathione peroxidase,and the expression of Bcl-2 and SIRT1 proteins in the model group were decreased,while the apoptosis rate,the relative content of reactive oxygen species,the level of malondialdehyde,the expression of Bax,matrix metalloproteinase 3,matrix metalloproteinase 9 proteins,and p-mTOR/mTOR were increased(P<0.05).Compared with the model group,the survival rate of chondrocytes,the positive rate of Edu,the levels of superoxide dismutase,catalase,and glutathione peroxidase,and the expression of Bcl-2 and SIRT1 proteins in the low-and high-dose emodin groups were increased,while the apoptosis rate,the relative content of reactive oxygen species,the level of malondialdehyde,the expression of Bax,matrix metalloproteinase 3,matrix metalloproteinase 9 proteins,and p-mTOR/mTOR were decreased(P<0.05).Compared with the low-and high-dose emodin groups,the indexes of EX527 group showed the opposite trend(P<0.05).Compared with the high-dose emodin group,the survival rate of chondrocytes,the positive rate of Edu,the levels of superoxide dismutase,catalase,and glutathione peroxidase,and the expression of Bcl-2 and SIRT1 proteins in the high-dose emodin+EX527 group were decreased,while the apoptosis rate,the relative content of reactive oxygen species,the level of malondialdehyde,the expression of Bax,matrix metalloproteinase 3,matrix metalloproteinase 9 proteins,and p-mTOR/mTOR were increased(P<0.05).To conclude,emodin can inhibit oxidative stress of osteoarthritic chondrocytes by activating the SIRT1-mTOR signaling,thereby promoting chondrocyte proliferation and reducing apoptosis.
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BACKGROUND:Knee osteoarthritis is a common clinical degenerative joint disease characterized by chronic inflammation and oxidative stress.Resveratrol has anti-inflammatory and anti-oxidative stress biological effects,and therefore it can be used symptomatically and expected to provide a new strategy for the treatment of knee osteoarthritis. OBJECTIVE:To investigate the therapeutic effect and mechanism of resveratrol on knee osteoarthritis in rats through the silence information regulator 1(SIRT1)/forkhead transcription factor O1(FOXO1)pathway. METHODS:Forty Sprague-Dawley rats were randomly divided into control group,model group,low-dose resveratrol group,and high-dose resveratrol group,with 10 rats in each group.Knee osteoarthritis models were established in the model group,low-dose resveratrol group,and high-dose resveratrol group.A mixture of 4%papain solution and 0.3 mol/L cysteine solution(1:1 for 0.5 hours;20 μL)was injected at 1,4,and 7 days after modeling.Rats in the low-dose and high-dose resveratrol groups were injected with 25 and 100 mg/kg resveratrol through the articular cavity at 1 day after successful modeling,while those in the control and model groups were injected with equivalent volume of physiological saline through the articular cavity.After 28 days of treatment,the maximum knee joint activity was measured;the levels of oxidative stress indicators and inflammatory factors in the synovial fluid of the knee joint were analyzed by radioimmunoassay and ELISA;the content of collagen fibers in the knee joint was analyzed by safranin O-fast green staining;the degree of arthritic lesions was analyzed using the Mankin histological score;and the levels of SIRT1 and FOXO1 in the knee joint were detected by western blot assay. RESULTS AND CONCLUSION:Compared with the model group,the maximum knee flexion and extension angles of rats significantly increased in the low-dose and high-dose resveratrol groups,and were significantly higher in the high-dose group than the low-dose group(P<0.05).Compared with the model group,the levels of superoxide dismutase and glutathione peroxidase in the knee joint fluid of rats significantly increased in the low-dose and high-dose resveratrol groups.The level of malondialdehyde significantly decreased in both resveratrol groups,and the level in the high-dose resveratrol group was significantly better than that in the low-dose resveratrol group(P<0.05).Compared with the model group,the low-dose and high-dose resveratrol groups showed a significant decrease in the levels of interleukin 1β,interleukin 6 and tumor necrosis factor α in the knee joint fluid of rats,and the levels of these inflammatory factors were significantly lower in the high-dose resveratrol group than the low-dose resveratrol group(P<0.05).Compared with the model group,the content of collagen fibers in the knee joint was significantly increased in both resveratrol groups,and the high-dose resveratrol group showed a higher content of collagen fibers than the low-dose resveratrol group(P<0.05).Compared with the model group,the expression level of SIRT1 in the knee joints of rats significantly increased in both resveratrol groups,while the level of acetylated FOXO1 significantly decreased(P<0.05).The magnitude of changes was significantly better in the high-dose group than the low-dose group.To conclude,resveratrol significantly improves the levels of oxidative stress and inflammatory factors in the joint fluid of rats with knee osteoarthritis and alleviates arthritic symptoms in a dose-dependent manner,possibly through the SIRT1/FOXO1 pathway.
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OBJECTIVE To explore the protective effects and potential mechanisms of Hirudo on mice with non-alcoholic fatty liver disease (NAFLD) in mice. METHODS The male ApoE-/- mice were randomly divided into the model group and Hirudo low- dose and high-dose groups (0.45, 0.9 g/kg), with 10 mice in each group; another 10 wild-type male C57BL/6J mice were chosen as the control group. The control group was fed with basal maintenance chow and the remaining groups were fed with high-fat chow for 12 weeks to establish the NAFLD model. Each administration group was given corresponding solution intragastrically, once a day, for 8 consecutive weeks. In the 13th week, the body weight and liver weight of mice in each group were measured after the last medication, and the liver index was calculated; the serum levels of nuclear factor-κB (NF-κB), tumor necrosis factor-α (TNF- α), interleukin-1β (IL-1β), IL-6, total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C) and high-density lipoprotein cholesterol (HDL-C) were detected; the liver pathomorphological changes were observed; the protein expressions of peroxisome proliferator-activated receptor γ(PPARγ) and silence information regulator type 1 (SIRT1) were detected. RESULTS Compared with the control group, the liver tissue of mice in the model group showed more fat vacuoles and infiltration of inflammatory cells, with significant lipid accumulation; the body weight, liver weight and liver index of the mice, and serum levels of NF-κB, TNF-α, IL-1β, IL-6, TC, TG and LDL-C significantly increased, while the serum level of HDL-C, the protein expressions of PPARγ and SIRT1 in liver tissues significantly decreased (P<0.01). Compared with the model group, the pathological changes in liver tissue of mice were all relieved in Hirudo low-dose and high-dose groups; the body weight, liver weight and liver index, the serum levels of NF-κB, TNF-α, IL-1β, IL-6, TC, TG and LDL-C decreased significantly, while the serum level of HDL-C, the protein expressions of PPARγ and SIRT1 in liver tissue all increased significantly (P<0.05 or P<0.01). CONCLUSIONS Hirudo can regulate liver lipid metabolism and inhibit inflammation by activating the protein expressions of PPARγ and SIRT1, thus having a significant ameliorative effect on NAFLD.
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ObjectiveTo explore the molecular mechanism of modified Shengjiangsan in alleviating endoplasmic reticulum (ER) stress and reducing urinary protein in the rat model of diabetic nephropathy (DN). MethodSeventy-five SD rats were randomized into normal, model, low-, medium-, and high-dose (4.37, 8.73, 17.46 g·kg-1, respectively) modified Shengjiangsan, and irbesartan (0.014 g·kg-1) groups, with 10 rats in each group. Rats were administrated with corresponding doses of medications or distilled water by gavage, once a day, for 8 consecutive weeks. After the last administration, the levels of glucose (GLU) in the blood, 24-hour urinary protein (24 h-UTP), and superoxide dismutase (SOD), malondialdehyde (MDA), and glutathione peroxidase (GSH-Px) in the renal tissue were measured. Hematoxylin-eosin staining, periodic acid-Schiff staining, and transmission electron microscopy were employed to observe the pathological changes in rat kidneys. Immunohistochemistry was employed to measure the expression levels of nephrin, podocin, glucose-regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), and activating transcription factor 4 (ATF4) in the kidneys of rats. Western blot was employed to measure the protein levels of silent information regulator 1 (Sirt1), phosphorylated (p)-protein kinase RNA-like endoplasmic reticulum kinase (PERK), and p-eukaryotic translation initiation factor 2 alpha (eIF2α) in rat kidneys. ResultCompared with the normal group, the modeling caused pathological damage to the kidneys, elevated the levels of GLU and 24 h-UTP (P<0.05), up-regulated the protein levels of GRP78, CHOP, ATF4, p-PERK, and p-eIF2α (P<0.05), and down-regulated the protein level of Sirt1 (P<0.05) in rat kidneys. Compared with the model group, modified Shengjiangsan and irbesartan lowered the GLU and 24 h-UTP levels (P<0.05), alleviated the pathological damage in the renal tissue, down-regulated the protein levels of GRP78, CHOP, ATF4, p-PERK, and p-eIF2α (P<0.05), and up-regulated the protein level of Sirt1 (P<0.05). ConclusionModified Shengjiangsan up-regulates Sirt1 expression and inhibits phosphorylation of proteins in the PERK/eIF2α pathway to reduce ER stress and oxidative stress in the renal tissue, thus alleviating the pathological damage in the renal tissue and reducing urinary protein in DN rats.
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AIM: To investigate the protective effect of Jianpi Bushen Yishi Formula on dry age-related macular degeneration(dARMD)induced by sodium iodate in mice and its mechanism.METHODS: A total of 27 SPF male C57BL/6 mice were randomly divided into blank, model and traditional Chinese medicine groups, with 9 in each group, the structure and morphology of the retina were observed by Hematoxylin-Ehong(HE)staining, and the intracellular reactive oxygen species(ROS)in the retina were observed by fluorescence staining with dihydroethidium(DHE). In addition, malondialdehyde(MDA)and superoxide dismutase(SOD)expression levels in mouse retina were detected by biochemical kit, and expression levels of silent information regulator type 1(SIRT1)and peroxisome proliferator-activated receptor-γ coactivator 1-α(PGC-1α)protein in mouse retina were detected by Western blot.RESULTS: Retinal structure and morphology of the model group showed a slight or mild decrease in the number of cells in the outer nuclear layer, a localized thinning of the outer nuclear layer, an inconspicuous demarcation between the outer and outer membranes, a slight or mild swelling of retinal pigment epithelial cells, and a slight or mild disturbance in the arrangement of retinal cells; while retinal pigment epithelial cells and photoreceptor layers in the traditional Chinese medicine group were significantly improved. DHE staining fluorescence results showed that the ROS level in the model group was significantly higher than that in the blank group at 14 d after modeling(P<0.01); the ROS level in the traditional Chinese medicine group was significantly lower than that in the model group(P<0.001). ELISA showed that the SOD level of the model group was significantly lower than that of the blank group at 14 d after modeling(P<0.01), and the MDA level was significantly increased(P<0.01)in the model group compared with the blank group; the SOD level was significantly higher(P<0.01), and the MDA level was significantly lower(P<0.01)in the traditional Chinese medicine group compared with the model group. Western blot results showed that the expression of SIRT1 and PGC-1α in the model group was significantly lower compared with that in the blank group(P<0.01), and the expression of SIRT1 and PGC-1α in the traditional Chinese medicine group was significantly higher compared with that in the model group at 7 and 14 d after modeling(P<0.01).CONCLUSION: The Chinese herbal medicine, which strengthens the spleen, tonifies the kidney and benefits the eyesight, can improve the oxidative stress state of the retina induced by sodium iodate in mice and reduce the damage to the retinal tissues, which may exert the anti-oxidative stress effect through the PGC-1α/SIRT1 signaling pathway.
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ObjectiveTo explore the underlying mechanism by which the Chinese medicine compound Yitangkang granule(YTK) treats diabetic kidney disease (DKD) by observing its effects on podocyte autophagy through the regulation of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/forkhead transcription factor O1 (FoxO1) signaling pathway mediated by silent information regulator 1 (SIRT1) via advanced glycation end products (AGE)/receptor for AGE (RAGE) axis. MethodNinety-six 8-week-old healthy male SPF-grade Wistar rats were selected and randomly divided into blank control group (B), model control group, high-dose YTK (40 g·kg-1), medium-dose YTK (20 g·kg-1), low-dose YTK (10 g·kg-1), and Western medicine control (20 mg·kg-1 losartan) groups. The DKD rat model was established by high-fat diet feeding combined with intraperitoneal injection of streptozotocin. After successful modeling, the rats in each group received the corresponding treatments for eight weeks. The levels of superoxide dismutase (SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-Px), and catalase (CAT) were measured according to the instructions of the respective assay kits. Hematoxylin and eosin (HE) staining was used to observe pathological changes in kidney tissues. Immunohistochemistry was employed to detect the average optical density values of α-smooth muscle actin (α-SMA), fibronectin (FN), desmin, and nephrin. Western blot analysis was used to measure the expression levels of PI3K, phosphorylated PI3K (p-PI3K), Akt, phosphorylated Akt (p-Akt), RAGE, SIRT1, Caspase-3, and FoxO1 proteins in kidney tissues of DKD rats. ResultCompared with the blank control group, the model group showed significantly lower levels of SOD, GSH-Px, and CAT, and significantly higher levels of MDA (P<0.01). The rats exhibited severe kidney damage. The positive expression of podocyte marker proteins α-SMA, FN, and desmin increased significantly, while nephrin and podocin significantly decreased (P<0.01). The expression levels of PI3K, p-PI3K, Akt, p-Akt, RAGE, and Caspase-3 proteins were significantly elevated, while SIRT1 and FoxO1 protein levels were significantly reduced (P<0.01). Compared with the model control group, rats in the YTK treatment groups showed significantly higher levels of SOD, GSH-Px, and CAT, and significantly lower levels of MDA in serum (P<0.01). The degree of kidney damage was reduced to varying extents. The average optical density values of podocyte marker proteins α-SMA, FN, and desmin were significantly decreased, while nephrin and podocin significantly increased (P<0.01). The expression levels of PI3K, p-PI3K, Akt, p-Akt, RAGE, and Caspase-3 in kidney tissues were significantly reduced, while SIRT1 and FoxO1 expression levels significantly increased (P<0.01). The Chinese medicine groups demonstrated a clear dose-response trend. ConclusionYTK may alleviate kidney pathological damage, reduce proteinuria, and protect kidney function in DKD rats, thereby delaying the progression of DKD by improving podocyte autophagy through the AGE-RAGE axis-mediated SIRT1 regulation of the PI3K/Akt/FoxO1 signaling pathway. Additionally, a dose-response relationship was observed in the Chinese medicine groups.
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OBJECTIVE To explore the neuroprotective effect of sodium aescinate on rats with Parkinson’s disease by regulating the silent information regulator 1 (SIRT1)/nuclear factor-κB (NF-κB) signaling pathway. METHODS The Parkinson’s disease rat model was constructed by using 6-hydroxydopamine injection method. Forty-eight rats successfully modeled were randomly divided into model group, sodium aescinate low-dose group (1.8 mg/kg), sodium aescinate high-dose group (3.6 mg/kg), sodium aescinate+EX527 (sodium aescinate 3.6 mg/kg+SIRT1 inhibitor EX527 5 mg/kg) group, with 12 rats in each group. Another 12 healthy rats were selected as the sham operation group. Each group was injected with the corresponding drug solution intraperitoneally, once a day, for 21 consecutive days. Twenty-four hours after the end of the last administration, the motor and cognitive functions of rats were detected, and the morphology of neurons in the substantia nigra and CA1 region of hippocampal tissue were observed. The content of dopamine (DA) in the nigrostriatal and the expression levels of tyrosine hydroxylase (TH) and α-synuclein (α-Syn) in the substantia nigra were detected. The serum levels of pro-inflammatory factor [interleukin-6 (IL-6), IL-18], anti-inflammatory factor (IL-10), and the expression levels of SIRT1, phosphorylated NF-κB p65 (p-NF-κB p65) and NF- κB p65 protein in nigrostriatal were detected. RESULTS Compared with sham operation group, the neurons in the substantia nigra and CA1 region of hippocampal tissue were seriously damaged in model group; the number of rotations, escape latency, the expression levels of α-Syn in substantia nigra, the levels of serum pro-inflammatory factors, the relative expression ratio of p-NF- κB p65 and NF-κB p65 protein in nigrostriatal were increased or prolonged significantly (P<0.05); the target quadrant residence time, the content of DA in nigrostriatal, the expression level of TH in substantia nigra, the serum level of anti-inflammatory factor, and the expression level of SIRT1 protein in substantia nigra striatum were significantly decreased or shortened (P<0.05). Compared with model group, the damage degrees of neuron in sodium aescinate groups were alleviated, and the quantitative indicators were significantly improved, which were more significant in the high-dose group (P<0.05); EX527 could reverse the improvement effect of high-dose sodium aescinate (P<0.05). CONCLUSIONS Sodium aescinate can inhibit the activation of NF-κB signal by up-regulating the protein expression of SIRT1, thereby reducing the neuroinflammation of rats with Parkinson’s disease, improving the motor and cognitive dysfunctions, and finally playing a neuroprotective role.
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OBJECTIVE To explore the improvement mechanism of proanthocyanidins on acute kidney injury (AKI) induced by gentamicin in rats. METHODS Gentamicin sulfate was injected intraperitoneally to construct the AKI rat model; the model rats were randomly divided into model control group, benazepril hydrochloride 5 mg/kg group (positive control), proanthocyanidins 50 mg/kg group, proanthocyanidins 100 mg/kg group, and proanthocyanidins 200 mg/kg group, with 10 rats in each group; in addition, 10 normal rats were selected to be treated as the normal control group. The rats in each administration group were given corresponding liquid intragastrically, and the normal control group and model control group were given equal volumes of normal saline intragastrically, once a day, for 28 consecutive days. After the last administration, the levels of serum creatinine (SCr), blood urea nitrogen (BUN), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and 24 h urinary protein (UP) were detected; the renal index was calculated; the pathological changes of renal tissue were observed and the pathological score was calculated; the apoptotic rate of cells in renal tissue and the expression levels of Caspase-3 and Bcl-2 associated X protein (Bax), as well as the phosphorylation levels of silent information regulator of transcription 1 (SIRT1) and AMP-activated protein kinase (AMPK) were detected. RESULTS Compared with the model control group, the levels of SCr, BUN, UP and MDA, the renal index, the pathological score of renal tissue, the apoptotic rate of cells in renal tissue, the protein expression levels of Caspase-3 and Bax in renal tissue of rats in each administration group were decreased significantly; SOD and GSH-Px levels, phosphorylation levels of SIRT1 and AMPK protein were increased significantly (P<0.05), and the effect of proanthocyanidins was in a dose-dependent manner (P<0.05). There were no significant differences in the above indexes between proanthocyanidins 200 mg/kg group and benazepril hydrochloride 5 mg/kg group (P>0.05). CONCLUSIONS The improvement effect of proanthocyanidins on AKI rats may be related to the activation of SIRT1/AMPK signaling pathway to inhibit oxidative stress.
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Aim To explore the role of SIRT1/Nrf2 / HO-1 in alleviating the cognitive function impairment by sevoflurane treatment in a mouse model of postoperative cerebral reperfusion. Methods C57BL/6J mice were randomly divided into five groups: sham operation group, hemorrhagic shock reperfusion group, sevoflurane postconditioning group, sevoflurane postcondition-ing + SIRT1 inhibitor group and sevoflurane postconditioning + Nrf2 inhibitor group. Mice were subjected to Morris water maze test after cerebral ischemia reperfusion. The ATP, superoxide dismutase (SOD), ROS and MDA contents in tissue of mice were detected. SIRT1, Nrf2 and HO-1 proteins in tissue were detected by Western blot. Results After hemorrhagic shock, the learning and memory ability of mice was reduced.ATP and SOD concentration in hippocampus was reduced , MDA and ROS concentration increased, and the SIRT, Nrf2 and HO-1 concentration was reduced. Sevoflurane improved the cognitive dysfunction and oxi-dative damage in postoperative mice, and the neuro-protective effect of sevoflurane on hemorrhagic shock and resuscitation mice was weakened followed with SIRT1 and Nrf2 inhibitors. Conclusion Sevoflurane probably alleviates the oxidative reaction damage and cognitive impairment caused by cerebral reperfusion in mice through SIRT1/Nrf2/H0-1 pathway.
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ObjectiveTo investigate the promotional effect of astragaloside on the repair and healing of chronic non-healing wounds and its mechanism. MethodA total of 60 male SD rats were constructed with full-layer skin defect wounds on the back, and except for the control (Con) group, the rest were constructed with non-healing wounds, which were then randomly divided into the sham-operation (sham) group, the low-dose astragaloside group, the high-dose astragaloside group, the astragaloside + LY294002 [phosphatidylinositol 3-kinase (PI3K) inhibitor] group, and the astragaloside + EX527 [silencing regulatory protein 1 (SIRT1) inhibitor] group. The percentage of wound area in each group was observed on the 2nd, 4th, 6th, and 8th days after wound molding. Collagen type Ⅰ alpha 1 (COL1A1) and alpha smooth muscle actin (α-SMA) expressions in the wound tissue were detected by immunofluorescence. Hematoxylin and eosin (HE) staining was performed to determine the pathological structure of the wound. The mRNA expression of inflammatory factors in the wound was measured by real-time polymerase chain reaction (Real-time PCR), and the expression of proteins related to the SIRT1/ nuclear factor (NF)-κB and PI3K/protein kinase B (Akt) signaling pathways in the wound was tested by Western blot. ResultCompared with the sham group, the percentage of postoperative wound area of rats in both low-dose and high-dose astragaloside groups gradually decreased with time, and the efficacy of the high-dose astragaloside group was better. Compared with the Con group, the fluorescence intensity of COL1A1 in wound tissue of the sham group decreased, while the expression of α-SMA increased. The epithelial tissue was severely damaged, with an increase in the thickness, and a large number of inflammatory cells were seen in the infiltration. The mRNA expression of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and inducible nitric oxide synthase (iNOS) was elevated. The protein expression of NF-κB p65, p-PI3K/PI3K, and p-Akt/Akt was elevated, while SIRT1 expression was decreased (P<0.05). Compared with the sham group, the fluorescence intensity of COL1A1 and α-SMA increased after astragaloside treatment. The number of epithelial cells increased, and the thickness decreased. The inflammatory cells decreased, and the amount of collagen increased. The mRNA expression of TNF-α, IL-1β, IL-6, and iNOS was decreased, and the protein expression of NF-κB p65, p-PI3K/PI3K, and p-Akt/Akt was decreased. SIRT1 was elevated, and the effect was better in the high-dose astragaloside group (P<0.05). Compared with the high-dose astragaloside group, inhibition of the PI3K/Akt and SIRT1 pathways by LY294002 and EX527 prevented the therapeutic efficacy of astragaloside on chronic non-healing wounds. ConclusionThe topical application of astragaloside significantly promotes the healing of chronic non-healing wounds in rats, and the mechanism may be related to the activation of the PI3K/Akt pathway and the SIRT1/NF-κB pathway.
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Objective:To investigate effect of acacetin on alveolar epithelial cell damage caused by Streptococcus pneumoniae(SP)infection by regulating sirtuin 1(Sirt1)-mediated 5'-AMP activated protein kinase(AMPK)/nuclear factor erythroid-2 related factor 2(Nrf2)signaling pathway.Methods:Alveolar epithelial cells A549 cultured in vitro were infected with SP to establish a cell damage model.After treatment with acacetin at final concentrations of 0,5,25,50,100,150,200 μmol/L,CCK-8 was performed to detect cell viability of each treatment group and optimal concentration of acacetin was screened.A549 cells cultured in vitro were ran-domly separated into five groups:control group,model group,acacetin(150 μmol/L)group,EX527(Sirt1 inhibitor,40 μmol/L)group,acacetin(150 μmol/L)+EX527(40 μmol/L)group,control group was not treated,other groups were infected with SP to establish a cell damage model,and then treated with 150 μmol/L acacetin and 40 μmol/L EX527,CCK-8 and flow cytometry were performed to measure cell viability and apoptosis rate in each group;kits were performed to measure levels of reactive oxygen species(ROS),superoxide dismutase(SOD),malondialdehyde(MDA),lactate dehydrogenase(LDH)and IL-10,IL-1β,TNF-α levels of cells in each group;Western blot was performed to measure proliferation-related proteins Ki-67,proliferating cell nuclear antigen(PCNA),apoptosis-related proteins caspase-9,Bax,Sirt1 and AMPK/Nrf2 signaling pathway proteins p-AMPK/AMPK,Nrf2 expres-sions of cells in each group.Results:Model group had decreased A549 cell viability,SOD and IL-10 levels,p-AMPK/AMPK,Sirt1,Nrf2,Ki-67 and PCNA protein expressions(P<0.05),and increased apoptosis rate,MDA,LDH,ROS,IL-1β and TNF-α levels than control group(P<0.05).Compared with model group and acacetin+EX527 group,acacetin group had increased A549 cell viability,SOD and IL-10 levels,p-AMPK/AMPK,Sirt1,Nrf2,Ki-67 and PCNA protein expressions(P<0.05),and decreased apoptosis rate,MDA,LDH,ROS,IL-1β and TNF-α levels(P<0.05);EX527 group had decreased A549 cell viability,SOD and IL-10 levels,p-AMPK/AMPK,Sirt1,Nrf2,Ki-67 and PCNA protein expressions(P<0.05),and increased apoptosis rate,MDA,LDH,ROS,IL-1β and TNF-α levels(P<0.05).Conclusion:Acnestin can activate AMPK/Nrf2 signaling by up-regulating Sirt1 expression,thereby promoting secretion of anti-inflammatory factors,reducing production of ROS and pro-inflammatory factors,reducing inflammation and oxidative stress,and finally alleviating neuronal damage.
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Objective This study aimed to explore the effect and possible mechanism of SIRT1 activation induced by butein on sciatic nerve injury in rats.Methods A total of 30 rats were randomly divided into a sham operation group,a sciatic nerve injury group,and a butein group,with 10 rats in each group.BBB motor scores and sciatic nerve function index were detected on the modeling surgery day,the 7th day after surgery,and the 14th day after surgery.The pathological changes of the sciatic nerve in each group were observed by HE staining.The apoptosis of sciatic nerve cells in each group was detected by TdT-mediated dUTP nick end labeling(TUNEL).The expres-sion of BDNF,MBP,GAP-43,SIRT1,FOXO1,Keap1,and NF-κB in the sciatic nerve was detected by Western blotting.Results Butein improved the pathological injury of the sciatic nerve,reduced the apoptosis of sciatic nerve cells,increased BDNF,MBP,GAP-43,and SIRT1 expression,and decreased FOXO1,Keap1,and NF-κB expression in the sciatic nerve.Conclusion Butein can inhibit FOXO1/NF-κB signaling pathway activation by up-regulating SIRT1 expression in rats with sciatic nerve injury and then improve the sciatic nerve pathological injury in rats.
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AIM: To detect the expression levels of long non-coding RNA(lncRNA)X-inactive specific transcript(XIST)and silencing information regulatory factor 2 associated enzyme 1(SIRT1)in serum of patients with type 2 diabetes mellitus(T2DM), and to explore their correlation with diabetic retinopathy(DR)and their diagnostic value. METHODS: Prospective study. A total of 214 patients with T2DM admitted to our hospital from January 2022 to February 2023 were selected as the research subjects. Based on whether retinopathy occurred, they were divided into 126 cases(126 eyes)in the non-DR group and 88 cases(88 eyes)in the DR group. An additional 130 healthy individuals who underwent a physical examination during the same period were selected as the control group. The serum levels of lncRNA XIST and SIRT1 in the three groups were measured and compared. The relationship between lncRNA XIST and SIRT1 expression with DR was analyzed using Pearson's method. The receiver operating characteristic(ROC)curve was used to evaluate the predictive value of serum lncRNA XIST, SIRT1, and their combination for DR. Multivariate Logistic regression analysis was performed to investigate the factors affecting the occurrence of DR in T2DM patients.RESULTS: Compared with the control group, the levels of serum lncRNA XIST and SIRT1 in the non-DR group and DR group were successively decreased(all P<0.05). The levels of serum lncRNA XIST and SIRT1 were positively correlated in DR patients(r=0.639, P<0.05). ROC analysis showed that the area under the curve(AUC)for predicting DR by combining serum lncRNA XIST and SIRT1 was 0.940, which was higher than the AUC by serum lncRNA XIST and SIRT1 alone(0.855, 0.875). Logistic regression analysis showed that lncRNA XIST(OR=0.752)and SIRT1(OR=0.694)were influencing factors for the occurrence of DR(both P<0.01).CONCLUSION: The serum levels of lncRNA XIST and SIRT1 are both lower in DR patients, and the combination of lncRNA XIST and SIRT1 has a better assessment capacity for the occurrence of DR.