红景天苷通过HIF-1途径对缺氧诱导心肌细胞凋亡的抑制作用  

Protective effect of salidroside on cardiomyocyte against apoptosis induced by hypoxia via HIF-1 pathway

作　　者：张金平[1] 陈建宗[1] 刘安恒[2] 张娟[1] 贾新[1] 蒋蔚峰[1] 

机构地区：[1]第四军医大学西京医院中医药研究所中西医结合老年脑病研究室,陕西西安710033 [2]第四军医大学西京医院心脏内科,陕西西安710033

出　　处：《第四军医大学学报》2008年第12期1057-1060,共4页Journal of the Fourth Military Medical University

基　　金：军队中药新药专项资助(2006192)

摘　　要：目的:研究红景天苷(Sal)抑制缺氧诱导的心肌细胞凋亡作用并探讨其可能的分子机制.方法:将原代培养的心肌细胞分为6组:正常对照组、缺氧组、缺氧+不同剂量Sal预处理组(10,50,100 mg/L)和缺氧+10-7mol/L胰岛素组.比色法测定细胞存活率、细胞培养上清液乳酸脱氢酶(LDH),肌酸激酶(CK)活力;Hoechst 33258染色观察细胞形态学改变;细胞流式检测术、DNA-ladder测定细胞凋亡;免疫荧光染色、蛋白印迹检测HIF-1α表达.结果:与缺氧组比较,不同浓度Sal预处理组可增加细胞存活率(P<0.05);显著降低细胞培养上清液LDH,CK活力(P<0.01);减少Hoechst染色阳性细胞(P<0.01);流式细胞仪、DNA-ladder检测凋亡细胞比率降低.缺氧可激活HIF-1α的表达并诱导其转位,100 g/L Sal可明显增加HIF-1α蛋白的表达(P<0.01).结论:Sal可抑制缺氧诱导的心肌细胞的凋亡,该抑制作用具有剂量依赖性,其分子机制可能与激活HIF-1α的表达,诱导其转位有关.AIM： To investigate the inhibitory effect of salidroside （Sal） on cardiomyocyte apoptosis induced by hypoxia and its molecular mechanism. METHODS： Primarily cultured cardiomyocytes of neonatal rats were divided into 6 groups： control group, hypoxia group, hypoxia ＋ different concentrations of Sal pretreatment groups （ 10,50,100 mg/L） , and hypoxia ＋ 10^-7 mol/L insulin group. The cell viability and activities of lactate dehydrogenase（LDH） and creatin kinase（CK） were measured by colorimetric method. The morphologic changes in cardiomyocytes were observed with Hoechst 33258 staining. The apoptosis of cardiomyocytes was detected by flow cytometry and DNA fragmen- tation. The expression of HIF-1α was determined by immunofluo- rescence and Western Blot. RESULTS： Compared with hypoxia group, pretreatment with different concentrations of Sal significantly attenuated hypoxia induced cell viability loss and cell death （P 〈0.05） and dramatically decreased the activities of the LDH and CK （ P 〈 0.01 ）. The Hoechst staining positive cells were significantly decreased with 100 mg/L Sal pretreatment （ P 〈0.01 ）. Using flow cytometry, we observed that Sal decreased cell apoptosis induced by hypoxia in a dose-dependent manner. Hypoxia activated the expression of the HIF-1α protein and induced the translocation of HIF-1α. Pretreatment with 100 mg/L Sal significantly up-regulated the level of HIF-1α protein. CONCLUSION： Sal protects cardiomyoeytes against apoptosis induced by hypoxia in a dose-dependent manner. This protection is possibly mediated throughactivating HIF-1α protein expression and inducing the translation of HIF-1α.

关 键 词：红景天苷 心肌/细胞学 细胞低氧 细胞凋亡 缺氧诱导因子-1Α 

分 类 号：R258.5[医药卫生—中医内科学]