白藜芦醇通过SIRT1/FoxO1路径改善大鼠脑缺血/再灌注损伤机制研究
|
摘要
目的研究白藜芦醇(RES)通过沉默信息调节因子1/叉头转录因子O1(SIRT1/FoxO1)路径改善大鼠脑缺血/再灌注(CIR)损伤的机制。方法 40只SD大鼠随机分为4组,假手术(Sham)组、模型(CIR)组、白藜芦醇(RES)组,白藜芦醇(RES)+SIRT1抑制剂(EX527)组,采用改良线栓法构建大鼠右侧大脑中动脉栓塞模型,采用逆转录-聚合酶链反应(RT-PCR)法和Western blot法检测各组大鼠脑组织匀浆中白细胞介素(IL)-1β、IL-6、肿瘤坏死因子(TNF)-α、SIRT1及核转录因子(NF)-κB mRNA含量及蛋白表达变化。选择1、3、7、14 d四个时间点参照改良神经损伤严重程度评分(mNSS)量表对大鼠神经功能缺损进行评分。结果 RES组3、7、14 d神经功能缺损评分明显低于CIR组和RES+EX527组(P<0.01)。RES组FoxO1、IL-1β、IL-6、TNF-αmRNA及蛋白表达量明显低于CIR组和RES+EX527组(P<0.01),而RES组SIRT1 mRNA及蛋白表达量明显高于CIR组和RES+EX527组(P<0.01)。结论 RES可通过SIRT1/FoxO1路径降低炎症因子TNF-α、IL-1β释放和促进抗凋亡因子B淋巴细胞瘤-2(Bcl-2)的表达,从而改善大鼠CIR损伤。
Objective To investigate the protective effect of resveratrol(resveratrol,RES) on cerebral ischemia/reperfusion(CIR) injury in rats and its relationship with sirtuin-1/forkhead transcription factor O1(SIRT1/FoxO1) pathway.Methods Forty SD rats were randomly divided into 4 groups: sham group, model(CIR) group, resveratrol(RES) group, resveratrol(RES)+EX527 group. The model of right middle cerebral artery occlusion was established in rats. Reverse transcription-polymerase chain reaction(RT-PCR) and Western blot were used to detect interleukin(IL)-1β,IL-6,tumor necrosis factor(TNF)-α,SIRT1 and NF-κB mRNA content and protein expression changes in brain homogenate of each group. Rats with neurological deficits were scored by reference to the mNSS scale at four time points,1 d,3 d,7 d, and 14 d.Results The neurological deficit scores of the RES group were significantly lower than those of the CIR group and the RES + EX527 group( P < 0. 01). The mRNA and protein expressions of FoxO1,IL-1β,IL-6 and TNF-α in RES group were significantly lower than those in CIR group and EX527 group( P < 0. 01),while the expression of SIRT1 mRNA and protein in RES group was significantly higher than that in CIR group and RES + EX527 group( P < 0. 01). Conclusion Resveratrol can inhibit the expression of FoxO1,reduce the release of inflammatory factors TNF-α,IL-1β and promote the expression of anti-apoptotic factor Bcl-2 by activating SIRT1,thereby improving cerebral ischemia/reperfusion damage in rats.
Objective To investigate the protective effect of resveratrol(resveratrol,RES) on cerebral ischemia/reperfusion(CIR) injury in rats and its relationship with sirtuin-1/forkhead transcription factor O1(SIRT1/FoxO1) pathway.Methods Forty SD rats were randomly divided into 4 groups: sham group, model(CIR) group, resveratrol(RES) group, resveratrol(RES)+EX527 group. The model of right middle cerebral artery occlusion was established in rats. Reverse transcription-polymerase chain reaction(RT-PCR) and Western blot were used to detect interleukin(IL)-1β,IL-6,tumor necrosis factor(TNF)-α,SIRT1 and NF-κB mRNA content and protein expression changes in brain homogenate of each group. Rats with neurological deficits were scored by reference to the mNSS scale at four time points,1 d,3 d,7 d, and 14 d.Results The neurological deficit scores of the RES group were significantly lower than those of the CIR group and the RES + EX527 group( P < 0. 01). The mRNA and protein expressions of FoxO1,IL-1β,IL-6 and TNF-α in RES group were significantly lower than those in CIR group and EX527 group( P < 0. 01),while the expression of SIRT1 mRNA and protein in RES group was significantly higher than that in CIR group and RES + EX527 group( P < 0. 01). Conclusion Resveratrol can inhibit the expression of FoxO1,reduce the release of inflammatory factors TNF-α,IL-1β and promote the expression of anti-apoptotic factor Bcl-2 by activating SIRT1,thereby improving cerebral ischemia/reperfusion damage in rats.
引文
[1] Tilley B C,Lyden P D,Brott T G,et al.Total quality improvement method for reduction of delays between emergency department admission and treatment of acute ischemic stroke.The National Institute of Neurological Disorders and Stroke rt-PA Stroke Study Group[J].Arch Neurol,1997,54(12):1466-74.
[2] Chong Z Z,Wang S,Shang Y C,et al.Targeting cardiovascular disease with novel SIRT1 pathways[J].Future Cardiol,2012,8(1):89-100.
[3] 李传文,张嵘,侯亮,等.SIRT1/NF-κB通路参与白藜芦醇改善大鼠脑缺血再灌注损伤炎性反应[J].安徽医科大学学报,2018,53(1):6-9.
[4] 帕尔哈提·热西提,更·党木仁加甫,汪永新,等.改良线栓法制作大鼠局灶性脑缺血模型[J].新疆医科大学学报,2006,29(2):145-7.
[5] Zhao Q,Zhang C,Wang X,et al.(S)-ZJM-289,a nitric oxide-releasing derivative of 3-n-butylphthalide,protects against ischemic neuronal injury by attenuating mitochondrial dysfunction and associated cell death[J].Neurochem Int,2012,60(2):134-44.
[6] Lippi G,Franchini M E,Favaloro E J,et al.Moderate red wine consumption and cardiovascular disease risk:beyond the "French paradox"[J].Semin Thromb Hemost,2010,31(1):59-70.
[7] Qian C,Jin J,Chen J et al.SIRT1 activation by resveratrol reduces brain edema and neuronal apoptosis in an experimental rat subarachnoid hemorrhage model[J].Mol Med Rep,2017,16(6):9627-35.
[8] Annabi B,Lord-Dufour S,Vézina A,et al.Resveratrol targeting of carcinogen-induced brain endothelial cell inflammation biomarkers MMP-9 and COX-2 is Sirt1-independent[J].Drug Target Insights,2012,6:1-11.
[9] Ponugoti B,Dong G,Graves D T.Role of forkhead transcription factors in diabetes-induced oxidative stress[J].Exp Diabetes Res,2013,2012(3):939751.
[10] 林超,陈慎仁,傅玉才,等.大鼠胰岛B细胞长寿基因SIRT1与转录调节因子FOXO1表达及其相关性研究[J].中国实用内科杂志,2007,27(14):1135-8.
[11] Xiong S,Salazar G,Patrushev N,et al.FoxO1 mediates an autofeedback loop regulating SIRT1 expression[J].J Biol Chem,2011,286(7):5289-99.
[12] Liu X,Yang X,Han L,et al.Pterostilbene alleviates polymicrobial sepsis-induced liver injury:Possible role of SIRT1 signaling[J].Int Immunopharmacol,2017,49:50-9.
[2] Chong Z Z,Wang S,Shang Y C,et al.Targeting cardiovascular disease with novel SIRT1 pathways[J].Future Cardiol,2012,8(1):89-100.
[3] 李传文,张嵘,侯亮,等.SIRT1/NF-κB通路参与白藜芦醇改善大鼠脑缺血再灌注损伤炎性反应[J].安徽医科大学学报,2018,53(1):6-9.
[4] 帕尔哈提·热西提,更·党木仁加甫,汪永新,等.改良线栓法制作大鼠局灶性脑缺血模型[J].新疆医科大学学报,2006,29(2):145-7.
[5] Zhao Q,Zhang C,Wang X,et al.(S)-ZJM-289,a nitric oxide-releasing derivative of 3-n-butylphthalide,protects against ischemic neuronal injury by attenuating mitochondrial dysfunction and associated cell death[J].Neurochem Int,2012,60(2):134-44.
[6] Lippi G,Franchini M E,Favaloro E J,et al.Moderate red wine consumption and cardiovascular disease risk:beyond the "French paradox"[J].Semin Thromb Hemost,2010,31(1):59-70.
[7] Qian C,Jin J,Chen J et al.SIRT1 activation by resveratrol reduces brain edema and neuronal apoptosis in an experimental rat subarachnoid hemorrhage model[J].Mol Med Rep,2017,16(6):9627-35.
[8] Annabi B,Lord-Dufour S,Vézina A,et al.Resveratrol targeting of carcinogen-induced brain endothelial cell inflammation biomarkers MMP-9 and COX-2 is Sirt1-independent[J].Drug Target Insights,2012,6:1-11.
[9] Ponugoti B,Dong G,Graves D T.Role of forkhead transcription factors in diabetes-induced oxidative stress[J].Exp Diabetes Res,2013,2012(3):939751.
[10] 林超,陈慎仁,傅玉才,等.大鼠胰岛B细胞长寿基因SIRT1与转录调节因子FOXO1表达及其相关性研究[J].中国实用内科杂志,2007,27(14):1135-8.
[11] Xiong S,Salazar G,Patrushev N,et al.FoxO1 mediates an autofeedback loop regulating SIRT1 expression[J].J Biol Chem,2011,286(7):5289-99.
[12] Liu X,Yang X,Han L,et al.Pterostilbene alleviates polymicrobial sepsis-induced liver injury:Possible role of SIRT1 signaling[J].Int Immunopharmacol,2017,49:50-9.