活血荣络方调控PI3K/Akt通路减轻脑缺血再灌注损伤PC12细胞炎症反应  

Mechanism of Action of Huoxue Rongluo Decoction on Reducing P12 Cell Inflammatory Response in Cerebral Ischemia-reperfusion Injury

作　　者：曾富康 张宇星 高晓峰[2] 刘利娟[2] 陈瑶[2] 李中[2] 周德生[2] Zeng Fukang;Zhang Yuxing;Gao Xiaofeng;Liu Lijuan;Chen Yao;Li Zhong;Zhou Desheng(Hunan University of Traditional Chinese Medicine,Hunan Province Key Laboratory of Integrated Chinese and Western Medicine for Prevention and Treatment of Heart and Brain Diseases,Hunan,Changsha 410208,China)

机构地区：[1]湖南中医药大学,中西医结合心脑疾病防治湖南省重点实验室,湖南长沙410208 [2]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《中国中医急症》2022年第12期2074-2077,2127,共5页Journal of Emergency in Traditional Chinese Medicine

基　　金：国家自然科学基金项目(82104766);湖南省自然科学基金(2021JJ30521,2021JJ40424);湖南省科技厅科技创新平台与人才计划(2017SK4005);湖南省财政中医药项目名院工程(rsk-010-013/006-09);湖南省教育厅一般项目(20C1405);湖南省卫健委科研项目(202103071190);湖南省中医药管理局资助项目(202046);湖南中医药大学中西医结合一流学科开放基金(2020ZXYJH38,2020ZXYJH39);中国卒中学会脑血管病全程管理项目-启航基金。

摘　　要：目的探讨活血荣络方对脑缺血再灌注损伤炎症反应的调控机制。方法通过体外实验,将PC12细胞分为空白组、模型组、活血荣络方组及阳性药物组(丁苯酞)。CCK8法检测细胞活力,探索最佳造模时间及含药血清浓度;Western blotting及RT-qPCR法检测PI3K/Akt通路相关蛋白及mRNA的表达水平;ELISA法检测细胞上清液中白细胞介素-1β(IL-1β)、白细胞介素-18(IL-18)和肿瘤坏死因子-α(TNF-α)水平。结果体外实验表明,与对照组相比,OGD4h/R24h细胞活力下降超过50%(P<0.01);与模型组相比,10%活血荣络方血清组细胞活力恢复最佳(P<0.01);活血荣络方能上调PI3K/Akt通路相关蛋白及mRNA表达(P<0.01),降低细胞上清液中IL-1β、IL-18和TNF-α炎症因子水平(P<0.01)。结论活血荣络方可能通过激活PI3K/Akt通路抑制脑缺血再灌注损伤炎症反应。Objective:To explore the regulation mechanism of Huoxue Rongluo Decoction on inflammatory response in cerebral ischemia-reperfusion injury(CIRI). Methods:PC12 cells were divided into a blank group,a model group,a Huoxue Rongluo Decoction group and a positive drug group(butylphthalein)by in vitro experiments. CCK8 assay was used to determine the cell viability and optimal molding time and serum concentration;Western blotting and RT-qPCR were used to determine the expression levels of PI3K/Akt pathway related proteins and mRNA;ELISA was used to determine the levels of IL-1β,IL-18 and TNF-α in the cell supernatant.Results:In vitro experiments showed that compared with the control group,the cell viability of OGD4h/R24h decreased by more than >50%(P < 0.01);compared with the model group,the cell viability of the 10% Huoxue Rongluo Decoction serum group recovered best(P < 0.01);Huoxue Rongluo Decoction could up-regulate the expression of PI3K/Akt pathway-related proteins and mRNA(P < 0.01),and reduce the levels of IL-1,β,IL-18 and TNF-αinflammatory factors in the cell supernatant(P < 0.01). Conclusion:Huoxue Rongluo Decoction may inhibit the inflammatory response to CIRI by activating the PI3K/Akt pathway.

关 键 词：脑缺血再灌注损伤 PC12细胞 活血荣络方 炎症反应 大鼠 体外实验 

分 类 号：R743.31[医药卫生—神经病学与精神病学]