松香酸通过诱导自噬减轻AGEs诱导的心肌H9c2细胞凋亡和内质网应激反应  

Abietic acid alleviates AGEs-induced apoptosis and endoplasmic reticulum stress in cardiomyocytes via inducing autophagy

作　　者：刘芳[1] 刘纪[1] 张颖[1] LIU Fang;LIU Ji;ZHANG Ying(Department of Internal Medicine,Tianin Huanhu Hospital,Tianjin 300381,China)

机构地区：[1]天津市环湖医院内科,天津300381

出　　处：《中国病理生理杂志》2018年第8期1376-1382,共7页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.80571057)

摘　　要：目的:本文旨在探索松香酸(abietic acid,AA)对晚期糖基化终末产物(advanced glycosylation end products,AGEs)诱导的心肌H9c2细胞凋亡和内质网应激的影响及其相关机制。方法:H9c2细胞分为5组:对照组加入生理盐水处理24 h;AGEs处理组加入AGEs(100 mg/L)处理24 h;AGEs+AA(10、25和50μmol/L)组同时加入AGEs(100 mg/L)和AA(10、25和50μmol/L)处理24 h。MTT法检测H9c2细胞活力。Western blot分析肌红蛋白(myoglobin,Mb)、肌酸激酶MB同工酶(creatine kinase MB isoenzyme,CK-MB)、心肌肌钙蛋白I(cardiac troponin I,c Tn I)、C/EBP同源蛋白(C/EBP homologous protein,CHOP)、cleaved caspase-12、GADD34、Bi P、LC3、P62和beclin 1的蛋白水平。ELISA检测乳酸脱氢酶(lactate dehydrogenase,LDH)活性。流式细胞术分析细胞凋亡。结果:低浓度(低于50μmol/L)的松香酸对H9c2细胞活力无明显影响,高浓度(高于50μmol/L)的松香酸会降低H9c2细胞活力。AGEs组Mb、CK-MB和c Tn I蛋白表达及LDH的水平明显高于对照组(P<0.05);与AGEs组相比,AGEs+AA(10、25和50μmol/L)组Mb、CK-MB和c Tn I的蛋白表达及LDH的表达水平明显降低(P<0.05)。松香酸(10、25和50μmol/L)可抑制AGEs诱导的心肌细胞凋亡、CHOP和cleaved caspase-12蛋白水平的升高及GADD34和Bi P表达的降低(P<0.05)。而且,松香酸(10、25和50μmol/L)可抑制AGEs诱导的心肌细胞LC3-Ⅱ/LC3-Ⅰ比值和beclin 1表达的降低及P62表达的升高(P<0.05)。自噬抑制剂3-甲基腺嘌呤可逆转松香酸对心肌细胞LC3、Mb、cleaved caspase-12和Bi P蛋白水平的影响(P<0.05)。结论:松香酸通过诱导自噬减轻AGEs诱导的心肌H9c2细胞凋亡和内质网应激反应。AIM： This study aims to explore the effect of abietic acid（ AA） on advanced glycosylation end products（ AGEs）-induced apoptosis and endoplasmic reticulum stress in H9 c2 cardiomyocytes. METHODS： H9 c2 cells were divided into 5 groups. The cells in control group were treated with saline for 24 h. The cells in AGEs treatment group were treated with AGEs（ 100 mg/L） for 24 h. The cells in AGEs ＋ AA（ 10,25 and 50 μmol/L） groups were simultaneously treated with AGEs（ 100 mg/L） and AA（ 10,25 and 50 μmol/L） for 24 h. The cell viability was measured by MTT assay. The protein levels of myoglobin（ Mb）,creatine kinase MB isoenzyme（ CK-MB）,cardiac troponin I（ c Tn I）,C/EBP homologous protein（ CHOP）,cleaved caspase-12,GADD34,Bi P,LC3,P62 and beclin 1 were determined by Western blot. The levels of lactate dehydrogenase（ LDH） were measured by ELASA. The apoptosis was analyzed by flow cytometry. RESULTS： The low concentration（ 50 μmol/L） of abietic acid had no obvious effect on the viability of H9 c2 cells. The high concentration（ 50 μmol/L） of abietic acid decreased the viability of H9 c2 cells. The levels of Mb,CK-MB,c Tn I and LDH in AGEs group were higher than those in control group（ P〈0. 05）. Compared with AGEs group,the levels of Mb,CK-MB,c Tn I and LDH in AGEs ＋ AA（ 10,25 and 50 μmol/L） groups were obviously reduced（ P〈0. 05）. Abietic acid at concentrations of 10,25 and 50 μmol/L inhibited AGEs-induced apoptosis,elevated the protein levels of CHOP and cleaved caspase-12,and attenuated expression of GADD34 and Bi P（ P〈0. 05）. Moreover,abietic acid at concentrations of 10,25 and 50 μmol/L suppressed AGEs-induced decreased ratio of LC3-Ⅱ/LC3-Ⅰ and expression of beclin 1,and enhanced the expression of P62（ P〈0. 05）. 3-Methyladenine,an inhibitor of autophagy,reversed the effect of abietic acid on the protein levels of LC3,Mb,cleaved caspase-12 and Bi P（P〈0.05）. CONCLUSION： Abietic acid alleviates AGEs-induced apoptosis a

关 键 词：松香酸 晚期糖基化终末产物 细胞凋亡 内质网应激 自噬 

分 类 号：R587.2[医药卫生—内分泌] R285.5[医药卫生—内科学]