苁蓉舒痉颗粒对帕金森病模型大鼠中脑线粒体氧化应激及自噬相关SIRT1/FoxO通路的影响  

Effect of Congrong Shujing Granules on Mitochondrial Oxidative Stress and Autophagy Related SIRT1/FoxO Pathway in Mesencephalon of Parkinson’s Disease Rats

作　　者：李茜羽 姚淑红 陈小倩 陈诗雅 王林[2] 袁明洲 蔡晶 LI Xiyu;YAO Shuhong;CHEN Xiaoqian;CHEN Shiya;WANG Lin;YUAN Mingzhou;CAI Jing(The Third People's Hospital of Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350108,China;College of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou,Fujian 350122,China)

机构地区：[1]福建中医药大学附属第三人民医院,福建福州350108 [2]福建中医药大学中西医结合学院,福建福州350122

出　　处：《福建中医药》2022年第3期26-30,共5页Fujian Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(82074507);国家自然科学基金青年科学基金项目(81804156,81904263);福建省自然科学基金项目(2018J01882)。

摘　　要：目的 探讨苁蓉舒痉颗粒对帕金森病(PD)线粒体氧化应激及自噬相关SIRT1/FoxO通路的影响。方法 采用软件随机数字法将27只健康SD大鼠分为正常组9只、造模组18只,造模组采用颈背部皮下注射鱼藤酮葵花油乳化液建立PD模型,将造模成功的大鼠随机分为模型组和治疗组各9只,治疗组予苁蓉舒痉颗粒溶液灌胃,正常组、模型组予等量生理盐水灌胃,连续干预14 d。分别于灌胃第1、7、14天进行爬杆实验检测爬杆实验得分;灌胃第14天大鼠麻醉后取材,免疫组化法检测中脑酪氨酸羟化酶(TH)阳性细胞数,ELISA法检测中脑沉默信息调节因子1(SIRT1)、叉头蛋白O1(FoxO1)、FoxO3a蛋白水平,Western blot法检测中脑SIRT1、FoxO1、FoxO3a、微管相关蛋白轻链3-Ⅰ(LC3-Ⅰ)、LC3-Ⅱ蛋白表达,流式细胞术检测中脑活性氧(ROS)水平。结果 与正常组比较,模型组爬杆实验评分、ROS水平明显升高(P<0.01),TH阳性细胞数及SIRT1、FoxO1、FoxO3a、LC3-Ⅰ、LC3-Ⅱ蛋白表达明显降低(P<0.05,P<0.01)。与模型组比较,治疗组TH阳性细胞数及SIRT1、FoxO1、FoxO3a、LC3-Ⅰ、LC3-Ⅱ蛋白表达明显升高(P<0.05,P<0.01),爬杆实验评分、ROS水平明显降低(P<0.01)。结论 苁蓉舒痉颗粒能改善帕金森病大鼠行为障碍,降低中脑组织ROS水平,增加自噬相关LC3-Ⅱ蛋白表达,保护中脑多巴胺能神经细胞,其保护作用可能是通过调节线粒体氧化应激及自噬相关SIRT1/FoxO通路实现的。Objective: To explore the effect of Congrong Shujing Granules(CRSJ) on mitochondrial oxidative stress and autophagy related SIRT1/FoxO pathway in Parkinson’s disease(PD). Methods: Twenty-seven SD rats were randomly divided into normal group(n=9) and modeling group(n=18). Rats in the modeling group were subcutaneously injected with rotenone-sunflower oil emulsion into the back of the neck to establish the rat model of PD. The successfully modeled rats were randomly divided into model group and treatment group, with 9 rats in each group. The treatment group was given CRSJ by gavage, and the normal group and the model group were given the same amount of normal saline by gavage;the interventions were administered for fourteen days in all the groups. The Pole climbing experiments was carried out at 1, 7, and 14 days after intragastric administration. After 14 days, the number of tyrosine hydroxylase(TH) positive cells in the midbrain were detected by immunohistochemistry;ELISA and Western blot methods were used to detect the content of silent information regulator 1(SIRT1), forkhead box protein O1(FoxO1), FoxO3a,microtubule-associated protein light chain 3-Ⅰ(LC3-Ⅰ) and LC3-Ⅱ in midbrain;flow cytometry was used to detect the level of brain reactive oxygen species(ROS). Results: Compared to those of the normal group, the pole climbing test score and ROS level in the model group were significantly higher(P<0.01), and the number of TH positive cells and the protein expression of SIRT1,FoxO1, FoxO3a, LC3-Ⅰ and LC3-Ⅱ were significantly lower(P<0.05). Compared with the model group, the number of TH positive cells and the protein expression of SIRT1, FoxO1, FoxO3a, LC3-Ⅰ and LC3-Ⅱ in the treatment group were significantly increased(P<0.05), and the score of pole climbing test and the level of ROS were significantly decreased(P<0.05). Conclusion: CRSJ can improve the behavior disorder of Parkinson’s disease rats, reduce the level of ROS in midbrain tissue, increase the expression of autophagy-related LC3-Ⅱ

关 键 词：帕金森病 苁蓉舒痉颗粒 线粒体自噬 氧化应激 SIRT1 FOXO 

分 类 号：R285.5[医药卫生—中药学]