毛蕊异黄酮苷通过SIRT1通路对缺血性脑损伤大鼠的保护作用及机制  

Protective Effect and Mechanism of Calycosin-7-O-β-D-glucoside through SIRT1 Pathway on Ischemic Brain Injury in Rats

作　　者：吴颖 王敏纯 廖卫国[1] 梁俊晖 王利胜[1] WU Ying;WANG Minchun;LIAO Weiguo;LIANG Junhui;WANG Lisheng(School of Pharmaceutical Sciences,Guangzhou University of Chinese Medicine,Guangzhou 510006 Guangdong,China)

机构地区：[1]广州中医药大学中药学院,广东广州510006

出　　处：《中药新药与临床药理》2022年第9期1163-1169,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(81573872,81873228);广东省普通高校重点领域专项(自然科学)项目(2021ZDZX2029)。

摘　　要：目的 探讨毛蕊异黄酮苷(CG)通过沉默信息调节因子 1(SIRT1)通路对缺血性脑损伤大鼠的保护作用及机制。方法 采用改良线栓法建立大脑中动脉闭塞(MCAO)大鼠模型;将大鼠随机分为 6 组:假手术组、模型组、尼莫地平组(20 mg·kg^(-1))、CG 组(40 mg·kg^(-1))、SIRT1 特异性抑制剂(EX527)组(10 mg·kg^(-1))、CG+EX527 组(40 mg·kgCG +10 mg·kgEX527),每组 13 只;尼莫地平、CG 灌胃给药,EX527 腹腔注射给药,每天 1 次,连续 7 d。参照 mNSS 评分标准对大鼠进行神经功能缺损评分;采用 TTC 染色法测定脑组织梗死体积;测定大鼠血清乳酸脱氢酶(LDH)、丙二醛(MDA)水平;Nissl、TUNEL 染色法观察大脑组织尼氏体及细胞凋亡情况;Western Blot法检测脑组织 SIRT1、Bcl-2 及 BAX 蛋白表达水平。结果 与模型组比较,CG 组大鼠第 3、5、7 天的 mNSS 评分明显下降(P<0.05,P<0.01);脑梗死体积明显减小(P<0.05);血清 LDH、MDA含量均明显降低(P<0.05,P<0.01);正常神经元的数量显著增加(P<0.01),尼氏体增多;细胞凋亡率明显降低(P<0.05);脑组织中的 BAX/Bcl-2 蛋白比值明显下降(P<0.05),SIRT1 蛋白表达明显升高(P<0.05)。与EX527 组比较,CG+EX527 组大鼠第 7 天的 mNSS 评分明显下降(P<0.05);脑梗死体积明显减小(P<0.05);血清 LDH、MDA 含量均明显降低(P<0.05,P<0.01);正常神经元的数量显著增加(P<0.01),尼氏体增多;细胞凋亡率显著降低(P<0.01);脑组织中的 BAX/Bcl-2 蛋白比值显著下降(P<0.05),SIRT1 蛋白表达显著升高(P<0.01)。结论 CG 对缺血性脑损伤大鼠具有保护作用,可能与调控 SIRT1 信号通路发挥抗氧化应激作用及减少脑组织细胞凋亡有关。Objective To investigate the protective effect and its mechanism of Calycosin-7-O-β-D-glucoside(CG)on ischemic brain injury in rats through silent information regulator 1(SIRT1) pathway. Methods The model of middle cerebral artery occlusion(MCAO)in rats was established by modified thread-occlusion method;the rats were randomly divided into 6 groups:sham-operation group,model group,Nimodipine group(20 mg·kg^(-1)),CG group(40 mg·kg^(-1)),SIRT1-specific inhibitor(EX527)group(10 mg·kg^(-1))and CG+EX527 group(40 mg·kgCG+ 10 mg·kgEX527),with 13 rats in each group. All the groups were administered with CG by gavage once a day for consecutive7 days. The rats were scored for transcutaneous functional deficits with reference to the mNSS scale;the infarct volume of brain tissue was detected by TTC staining;serum LDH and MDA levels were detected;Nissl and TUNEL staining were used to observe the brain tissue and apoptosis;the protein expression levels of SIRT1,Bcl-2 and BAX in brain tissues were detected by Western Blot. Results Compared with the model group,the mNSS scores of rats in the CG group on days 3,5 and 7 were significantly decreased(P<0.05,P<0.01);the brain infarct volume was significantly reduced(P<0.05);serum LDH and MDA levels were significantly reduced(P<0.05,P<0.01);the number of normal neurons was significantly increased(P<0.01)and Nissl substance was increased;apoptosis rate was significantly reduced(P<0.05);the BAX/Bcl-2 protein ratio in brain tissues was significantly decreased(P<0.05)and SIRT1 protein expression was significantly increased(P<0.05). Compared with the EX527 group,the m NSS scores of rats in the CG+EX527 group on day 7 were significantly decreased(P<0.05);the brain infarct volume was significantly reduced(P<0.05);serum LDH and MDA levels were significantly reduced(P<0.05,P<0.01);the number of normal neurons was significantly increased(P<0.01)and Nissl substance was increased;the apoptosis rate was significantly reduced(P<0.01);the BAX/Bcl-2 protein ratio in brain tissues was significan

关 键 词：毛蕊异黄酮苷 缺血性脑损伤 SIRT1信号通路 凋亡 抗氧化应激 大鼠 

分 类 号：R285.5[医药卫生—中药学]