补阳还五汤对脑缺血再灌注大鼠线粒体氧化损伤和PKCε-Nampt通路的影响  

Effects of Buyang Huanwu Decoction on mitochondrial oxidative damage and PKCε-Nampt pathway in rats following cerebral ischemia-reperfusion

作　　者：尹美娟 刘真一[1] 靳晓飞 周晓红[1] 高钰 赵月眸 高维娟[1] YIN Mei-juan;LIU Zhen-yi;JIN Xiao-fei;ZHOU Xiao-hong;GAO Yu;ZHAO Yue-mou;GAO Wei-juan(Hebei Key Lab of Chinese Medicine Research on Cardio-cerebrovascular Disease,Hebei University of Chinese Medicine,Shijiazhuang 050091,China)

机构地区：[1]河北中医学院,河北省心脑血管病中医药防治研究重点实验室,河北石家庄050091

出　　处：《中成药》2024年第1期79-86,共8页Chinese Traditional Patent Medicine

基　　金：中央引导地方科技发展资金项目(206Z7706G);河北省属高校基本科研业务费项目(YZYX2022003);河北中医学院科技能力提升重点项目(KTZ2019018);河北中医学院研究生创新资助项目(XCXZZSS2022013)。

摘　　要：目的探讨补阳还五汤对脑缺血再灌注(I/R)大鼠线粒体氧化损伤及PKCε-Nampt通路的影响。方法大鼠随机分为假手术组、模型组、补阳还五汤组(14.3 g/kg)和依达拉奉组(3 mg/kg),除假手术组外均采用MCAO法建立脑I/R损伤模型,术后24 h各组给予相应剂量药物,给药7 d后通过神经功能评分评估神经功能缺损情况,免疫荧光检测神经元标志物MAP-2表达,观察神经元损伤情况,检测ROS、MDA水平和SOD活性评价氧化损伤情况,荧光探针JC-1检测线粒体膜电位变化,修饰酶循环法检测NAD^(+)/NADH比值,Western blot法检测PKCε、p-PKCε、Nampt蛋白表达,免疫组化法检测p-PKCε、Nampt蛋白阳性表达。结果与模型组比较,补阳还五汤组脑梗死体积、神经功能评分降低(P<0.05),脑组织MAP-2荧光强度增强(P<0.05),神经元损伤减轻,脑组织ROS、MDA水平降低(P<0.05),SOD活性、线粒体膜电位、NAD^(+)/NADH比值升高(P<0.05),p-PKCε、Nampt蛋白表达升高(P<0.05)。结论补阳还五汤可通过激活PKCε-Nampt信号通路、提高线粒体功能来减轻大鼠脑I/R损伤。AIM To explore the effects of Buyang Huanwu Decoction on mitochondrial oxidative damage and PKCε-Nampt pathway in rats following cerebral ischemia reperfusion(I/R).METHODS The rats were randomly divided into the sham operation group,the model group,Buyang Huanwu Decoction group(14.3 g/kg)and edaravone group(3 mg/kg).Except those of the sham operation group,SD rats of other groups were induced into models of brain I/R injury by MCAO method,followed by corresponding drug administration 24 hours after operation.After 7 days of administration,the rats had their neurological deficit evaluated by neurological function scoring;thier expression of neuron marker MAP-2 detected by immunofluorescence staining;their neuron damage observed and the oxidative damage evaluated through assessment of their ROS levels and MDA and SOD activities;their changes of mitochondrial membrane potential detected by fluorescent probe JC-1;their ratio of NAD^(+)/NADH detected using modified enzyme circulation method;their expressions of PKCε,p-PKCεand Nampt proteins detected with Western blot;and their positive expressions of p-PKCεand Nampt proteins detected with immunohistochemistry method.RESULTS Compared with the model group,Buyang Huanwu Decoction group shared decreased cerebral infarction volume and neurological function score(P<0.05);increased cerebral fluorescence intensity of MAP-2(P<0.05);reduced neuronal damage,decreased cerebral levels of ROS and MDA(P<0.05);increased SOD activity,mitochondrial membrane potential and NAD^(+)/NADH ratio(P<0.05);and increased protein expressions of p-PKCεand Nampt(P<0.05).CONCLUSION Buyang Huanwu Decoction can improve mitochondrial function and reduce brain I/R injury in rats by activating their PKCε-Nampt signaling pathway.

关 键 词：补阳还五汤 脑缺血再灌注损伤 线粒体 PKCΕ NAMPT 

分 类 号：R285[医药卫生—中药学]