基于IRE1α/XBP1/CHOP通路探讨黄连解毒汤对内质网应激的作用机制  

Study on the Mechanism of Huanglian Jiedu Decoction on ERS Based on IRE1α/XBP1/CHOP Pathway

作　　者：周月 吴福佳 李建橡 张爱珍 贺鸿捷 许育佳 岳桂华[1] ZHOU Yue;WU Fujia;LI Jianxiang(Guangxi University of Chinese Medicine,Nanning Guangxi 530200,China;Ruikang hospital affiliated to Guangxi university of Chinese medicine,Nanning Guangxi 530011,China)

机构地区：[1]广西中医药大学,广西南宁530200 [2]广西中医药大学附属瑞康医院,广西南宁530011 [3]广西卫生职业技术学院,广西南宁530023

出　　处：《四川中医》2024年第2期86-91,共6页Journal of Sichuan of Traditional Chinese Medicine

基　　金：国家自然科学基金-地区科学基金项目(编号:81860831);广西中医药大学校级研究生科研创新项目(编号:YCSY2023058)。

摘　　要：目的:研究黄连解毒汤含药血清对衣霉素诱导的人脐静脉内皮细胞(Human Umbilical Vein Endothelial Cells,HUVEC)内质网应激的内质网跨膜激酶1(IRE1α)、X盒结合蛋白(XBP1)、环磷酸腺苷反应元件结合转录因子同源蛋白(C/EBP-homologous protein,CHOP)表达的影响,以探讨黄连解毒汤作用于内质网应激的细胞通路机制。方法:制备HLJDD和卡托普利含药血清,将细胞分为空白对照组、模型组、HLJDD低、中、高剂量组、卡托普利组。采用CCK8法检测细胞存活率;流式细胞术检测各组细胞凋亡水平。运用蛋白印记检测HUVEC的内质网应激葡萄糖调节蛋白78(glucose regulated protein 78kD,GRP78)、CHOP、IRE1α、XBP1、PDIA6、eNOS蛋白的表达变化;实时荧光定量PCR检测细胞的GRP78、CHOP、IRE1α、XBP1、PDIA6及eNOS的mRNA表达。结果:与空白对照组相比,模型组细胞存活率降低,凋亡率明显升高,GRP78、CHOP、IRE1α、XBP1、PDIA6表达升高(P<0.05);eNOS表达降低(P<0.05)。与模型组相比,黄连解毒汤各组和卡托普利干预后,细胞凋亡率明显降低,GRP78、CHOP、IRE1α、XBP1、PDIA6表达降低(P<0.05);eNOS表达升高(P<0.05)。结论:黄连解毒汤可改善ERS状态,抑制细胞凋亡,达到对内皮细胞的保护作用,其作用机制可能与抑制IRE1α/XBP1/CHOP信号通路有关。Objective To study the effect of Huanglian Jiedu Decoction(HLJDD)containing serum on endoplasmic reticulum transmembrane kinase 1(IRE1)in the endoplasmic reticulum stress of human umbilical vein endothelial cells(HUVEC)induced by tunicamycinα、The effect of X box binding protein(XBP1)and cyclic adenosine monophosphate response element binding transcription factor homologous protein(C/EBP homologous protein,CHOP)on the expression of X box binding protein(XBP1)was studied to explore the cellular pathway mechanism of Huanglian Jiedu decoction on endoplasmic reticulum stress.Methods HLJDD and captopril containing serum were prepared,and the cells were divided into control group,TM group,HLJDD low,medium and high dose groups,and captopril group.The cell survival rate was detected by CCK8method;The level of apoptosis was detected by flow cytometry.The protein expression of endoplasmic reticulum stress glucose-regulated protein 78kD(GRP78),CHOP,IRE1α,XBP1,PDIA6,eNOS was detected by protein blotting;and the mRNA of GRP78,CHOP,IRE1α,XBP1,PDIA6,eNOS was detected by real-time fluorescent quantitative PCR.Results Results Compared with blank control group,the cell survival rate in model group was decreased,the apoptosis rate was increased,and the expressions of GRP78,CHOP,IRE1α,XBP1and PDIA6were increased(P<0.05).The expression of eNOS was decreased(P<0.05).Compared with model group,the apoptosis rate and the expressions of GRP78,CHOP,IRE1α,XBP1and PDIA6were decreased significantly in Huanglian Jiedu decoction group and captopril group(P<0.05).eNOS expression was increased(P<0.05).Conclusion Huanglian Jiedu Decoction can improve ERS status,inhibit cell apoptosis,and protect endothelial cells.The mechanism may be related to the inhibition of IRE1α/XBP1/CHOP signaling pathway.

关 键 词：黄连解毒汤 内质网应激 内皮细胞 信号通路 

分 类 号：R285.5[医药卫生—中药学]