京尼平抑制饱和脂肪酸诱导的HepG2细胞凋亡和内质网应激  

Genipin inhibits apoptosis and endoplasmic reticulum stress induced by saturated fatty acid in HepG2 cell

作　　者：宋璐璐[1] 萧建中[2] 杨文英[2] 张敏[3] 柳彬彬[3] 

机构地区：[1]北京协和医学院研究生院,100071 [2]北京中日友好医院内分泌科,100029 [3]北京大学医学部研究生院,100091

出　　处：《国际内分泌代谢杂志》2011年第2期80-83,共4页International Journal of Endocrinology and Metabolism

基　　金：首都医学科研发展基金资助项目（2005-1036）

摘　　要：目的探讨京尼平减轻棕榈酸对HepG2细胞毒性的作用及机制。方法将HepG2细胞分为4组，分别用牛血清白蛋白、棕榈酸（1mmol／L）、京尼平（20μmol／L）或京尼平（20μmol／L）预处理30min后棕榈酸（1mmol／L）孵育24h，检测细胞活力及乳酸脱氢酶（LDH）释放；孵育16h后经流式细胞术和Hoechst染色检测细胞凋亡；孵育6h后实时荧光定量PCR检测糖调节蛋白（GRP）78、CCAAT增强子结合蛋白同源蛋白（CHOP）基因表达，PCR结合电泳检测x盒结合蛋白（XBP）-1剪接体。结果和空白组（牛血清白蛋白）相比，棕榈酸可降低HepG2细胞活力（P〈0．05）、增加LDH释放（P〈0．01）和HepG2细胞凋亡（P〈0．05），上调GRP78、CHOP的表达（P〈0．001）及XBP-1的剪接；和棕榈酸组相比，京尼平增加细胞活力（P〈0．05）、减少LDH释放（P〈0．05），显著抑制细胞凋亡（P〈0．01），降低GRP78、CHOP基因（P〈0．01）的表达。PCR产物电泳显示京尼平减少了XBP-1的剪接。结论京尼平对棕榈酸诱导的细胞凋亡有保护作用，其机制可能与抑制内质网应激有关。Objective To examine the protective effect of genipin from palmitate-indueed cytotoxicity in HepG2 cells and investigate the underlying mechanism. Methods HepG2 cells were divided into 4 groups and were treated respectively with bovine serum albumin （BSA）, pahnitate （1 mmoi/L）, genipin（20 μmol/L） or palmitate for 24 h after genipin pretreatment for 30 rain. Assayed the cell viability and lactate dehydrogenase enzyme （LDH） release. Fiowcytometry and Hoeehst staining were employed for determination of cell apoptosis after 16 h-treatment. Glucose-regulated protein（GRP） 78 and CCAAT enhancer binding protein-homologous protein（CHOP） mRNA expression was quantified by real time PCR while X-box binding protein（ XBP） -i splicing was showed by PCR and electrophoresis after 6 h-treatment. Results Compared with BSA, palmitate decreased cell viability （P 〈 0.05 ）while increased LDH release（P 〈 0. 01 ）. It also significantly induced apoptosis of HepG2 cell （P 〈 0.05 ）. Expression of GRP78 and CHOP mRNA was up-reg- ulated by palmitate （P 〈 0. 001 ）, so was XBP-1 splicing. Compared with palmitate, genipin pretreatment increased cell viability （ P 〈 0.05 ）, reduced LDH release （ P 〈 0.05 ） and inhibited apoptosis （ P 〈 0.01 ） of HepG2 cell. The expression of GRP78 and CHOP mRNA was decreased by genipin（P 〈 0.01 ）. Electrophoresis of XBP-1 PCR products showed less spliced XBP-1 in genipin-palmitate treated ceils than palmitate treated ones. Conclusion Genipin protects HepG2 cells from pahnitate-induced cell apoptosis, which may be mediated by inhibition of endoplasmic reticulum stress.

关 键 词：脂毒性 京尼平 细胞凋亡 内质网应激 

分 类 号：R285.5[医药卫生—中药学]