杨桃根DMDD对高糖诱导的肾小管上皮细胞HK-2内质网应激IRE1α通路及炎症反应的抑制作用  

The inhibitory effect of Averrhoa carambola DMDD on high glucose-induced endoplasmic reticulum stress IRE1α pathway and inflammation in renal tubular epithelial cell HK-2

作　　者：王誉湘 谢静晓 张晓萍 庞川皓 王璐 陈秋燕 陈林倩 黄仁彬[1] 韦晓洁 WANG Yu-xiang;XIE Jing-xiao;ZHANG Xiao-ping;PANG Chuan-hao;WANG Lu;CHEN Qiu-yan;CHEN Lin-qian;HUANG Ren-bin;WEI Xiao-jie(Pharmaceutical College,Guangxi Medical University,Nanning 530021,China;School of Basic Medine,Guangxi University of Traditional Chinese Medicine,Nanning 530021,China)

机构地区：[1]广西医科大学药学院,广西南宁530021 [2]广西中医药大学基础医学院,广西南宁530021

出　　处：《中国药理学通报》2023年第7期1270-1275,共6页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No.82004186,81760665);广西自然科学基金资助项目(No.2020GXNSFAA297181)。

摘　　要：目的研究杨桃根活性成分2-十二烷基-6-甲氧基-2,5-二烯-1,4-环己二酮(DMDD)对高糖诱导肾小管上皮细胞HK-2内质网应激及炎症反应的抑制作用。方法体外培养HK-2细胞,并分为正常组、高糖组、内质网应激抑制剂4-PBA组(5 mmoL·L^(-1))和DMDD高、中、低浓度组(8、4、2μmol·L^(-1))。各组细胞培养结束后,采用CCK-8法检测细胞存活率;ELISA检测细胞上清液中TNF-α、IL-6的水平;AO/EB染色法检测细胞凋亡;Western blot法检测细胞中凋亡相关蛋白Bcl-2、Bax的表达水平、内质网应激标志蛋白GRP78、CHOP的表达水平以及IRE1α通路相关蛋白(IRE1α、JNK)的磷酸化水平。结果与高糖组比较,DMDD各浓度组细胞存活率明显升高;TNF-α、IL-6水平,Bax/Bcl-2、GRP78、CHOP蛋白表达水平和IRE1α、JNK蛋白磷酸化水平均明显降低,细胞中橙红色荧光明显减少。结论DMDD可能部分通过抑制内质网应激及IRE1α通路相关蛋白的表达,减少炎症反应,达到HK-2细胞的保护作用。Aim To investigate the inhibition effect of 2-dodecyl-6-methoxycyclohexa-2,5-diene-1,4-dione(DMDD)on renal tubular epithelial cell HK-2 endoplasmic reticulum stress and inflammatory responses induced by high glucose.Methods HK-2 cells were cultured in vitro and divided into normal group,high glucose group,endoplasmic reticulum stress inhibitor 4-PBA group(5 mmoL·L^(-1)),DMDD high,medium and low dose groups(8,4,2μmol·L^(-1)).CCK-8 assay was used to measure cell viability after cells in each group were cultured.ELISA was used to detect the levels of TNF-αand IL-6 in cell supernatant.AO/EB staining was used to detect cell apoptosis.Western blot was used to detect the expression of apoptosis-related proteins(Bcl-2,Bax),endoplasmic reticulum stress markers(GRP78,CHOP)and phosphorylation levels of IRE1αpathway-related proteins(IRE1α,JNK).Results Compared with high glucose group,cell viability of DMDD groups significantly increased,levels of inflammatory factors TNF-αand IL-6 significantly decreased;Bax/Bcl-2,GRP78,CHOP,IRE1αand JNK protein expression levels markedly decreased,and range-red fluorescence was apparently reduced.Conclusions DMDD may inhibit the endoplasmic reticulum stress and the expression of IRE1αpathway-related proteins and reduce the inflammatory response,thus achieving the protective effect on high glucose-induced HK-2 cells.

关 键 词：2-十二烷基-6-甲氧基-2 5-二烯-1 4-环己二酮 糖尿病肾病 肾小管上皮细胞 内质网应激 凋亡 IRE1α通路 

分 类 号：R284.1[医药卫生—中药学] R322.61[医药卫生—中医学] R329.24R364.5R587.2