健脾益智法调控线粒体凋亡HtrA2/Omi通路保护MCAO大鼠脑神经的机制研究  

A study on the mechanism of Jianpi Yizhi in protecting brain nerve in MCAO rats via regulating mitochondrial apoptosis HtrA2/Omi pathway

作　　者：游雪娟 冯珂[1] 纪立金[1] 张子恒 吴佳琪 陈丽斌[1] YOU Xuejuan;FENG Ke;JI Lijin;ZHANG Ziheng;WU Jiaqi;CHEN Libin(The Medical College of Traditional Chinese Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China)

机构地区：[1]福建中医药大学中医学院,福州350122

出　　处：《环球中医药》2025年第1期7-12,共6页Global Traditional Chinese Medicine

基　　金：国家自然科学基金面上项目(81373528);福建省自然科学基金面上项目(2022J01873);福建中医药大学校管课题(X2021001-重点)。

摘　　要：目的观察健脾益智法对缺血性脑卒中大鼠线粒体丝氨酸蛋白酶(high temperature requirement factor A2/Omi,HtrA2/Omi)通路的影响,探讨其脑神经保护机制。方法将48只雄性SD大鼠随机分为正常组、模型组、健脾益智组(灌胃,14.4 mL/kg)和UCF101组(腹腔注射,1.5μmol/kg),每组12只,建立大鼠大脑中动脉栓塞(middle cerebral artery occlusion,MCAO)模型,各治疗组给药1次,各组24小时后取材。测定Zea Longa神经功能缺损评分及神经功能改善评分有效率。采用RT-qPCR检测HtrA2/Omi、X连锁凋亡抑制蛋白(X-linked inhibitor of apoptosis protein,XIAP)、半胱氨酸蛋白酶(Caspase-9、Caspase-3)、DNA修复酶(poly ADP-ribose polymerase,PARP)等基因表达水平;采用蛋白免疫印迹法(Western Blot,WB)检测HtrA2/Omi、XIAP、Cleaved Caspase-3、Cleaved Caspase-9、PARP等蛋白表达情况。结果(1)Zea Longa神经功能缺损评分结果:与正常组比较,MCAO模型各组大鼠神经功能评分显著升高(P<0.01)。(2)神经功能改善评分有效率:健脾益智组有效率100%,UCF101组有效率达58.3%(P<0.05)。(3)RT-qPCR结果:与正常组比较,模型组HtrA2/Omi、Caspase-3、PARP基因表达水平显著升高(P<0.05,P<0.01),XIAP基因表达无统计意义(P>0.05),Caspase-9趋势升高但无统计意义(P>0.05)。与模型组比较,健脾益智组、UCF101组HtrA2/Omi、Caspase-3、PARP基因表达水平显著降低(P<0.05,P<0.01),Caspase-9趋势降低但无统计意义(P>0.05);健脾益智组XIAP基因表达水平显著升高(P<0.01),UCF101组XIAP基因表达水平虽有所上升,但无统计意义(P>0.05)。(4)WB结果:与正常组比较,模型组HtrA2/Omi、Cleaved Caspase-3、Cleaved Caspase-9、PARP蛋白表达水平显著升高(P<0.01),XIAP蛋白表达水平显著下降(P<0.01)。与模型组比较,健脾益智组、UCF101组HtrA2/Omi、Cleaved Caspase-3、Cleaved Caspase-9、PARP蛋白表达水平显著降低(P<0.05,P<0.01),XIAP蛋白表达水平显著升高(P<0.01)。结论健脾益智法可能改善MCObjective To observe the effect of Jianpi Yizhi on the mitochondrial HtrA2/Omi apoptosis pathway in rats with ischemic stroke,and explore the brain nerve protection mechanism.Methods 48 SD rats were randomly divided into the normal group,the model group,the Jianpi Yizhi group(gavage,14.4 mL/kg)and the UCF101 group(intraperitoneal injection,1.5μmol/kg),with 12 rats in each group,middle cerebral artery embolization(MCAO)rat model,once per treatment group,24 hours after each group.Testing Zea Longa neurodeficit score and effective rate of nerve function improvement score.Gene expression levels of HtrA2/Omi,XIAP,Caspase-3,Caspase-9,and PARP were determined by RT-qPCR,and protein expression of HtrA2/Omi,XIAP,Cleaved Caspase-3,Cleaved Caspase-9,and PARP were determined by Western Blot.Results(1)Zea Longa neurodeficit score results:compared with the normal group,the neurological score of rats in the MCAO model was significantly increased(P<0.01).(2)Effective rate of nerve function improvement score:the effective rate was 100%,and 58.3%in the UCF101 group(P<0.05).(3)Results of RT-qPCR:compared with the normal group,the levels of HtrA2/Omi,Caspase-3 and PARP were significantly increased(P<0.05 or P<0.01),XIAP was not statistically significant,Caspase-9 trend was increased but not statistically significant.Compared with the model group,the expression levels of HtrA2/Omi,Caspase-3 and PARP were significantly decreased in the Jianpi Yizhi group and the UCF101 group(P<0.05 or P<0.01),and the trend of Caspase-9 was decreased but had not statistically significan(P>0.05).The expression level of XIAP gene in Jianpi Yizhi group was increased significantly(P<0.01),while the expression level of XIAP gene increased in the UCF101 group,but it had no statistical significance(P>0.05).(4)Results of WB:Compared with the normal group,the protein expression levels of HtrA2/Omi,Cleaved Caspase-3,Cleaved Caspase-9,and PARP were significantly increased(P<0.01),and the XIAP protein expression level was significantly decreased in the model grou

关 键 词：健脾益智法 细胞凋亡 线粒体 线粒体丝氨酸蛋白酶 缺血性脑卒中 

分 类 号：R285.5[医药卫生—中药学]