HO-1在LPS致大鼠肺泡巨噬细胞凋亡中的内源性保护作用:与内质网应激的关系  

Role of HO-1-induced endogenous protection in LPS-caused apoptosis in rat alveolar macrophages:relationship with endoplasmic reticulum stress

作　　者：胡欣欣 宫丽荣[2] 史佳[2] 吴丽丽[2] 李翠[1] 吴建华 赵丁欢 余剑波[2] Hu Xinxin;Gong Lirong;Shi Jia;Wu Lili;Li Cui;Wu Jianhua;Zhao Dinghuan;Yu Jianbo(Nankai Clinical College of Tianjin Medical University,Tianjin 300100,China;Department of Anesthesiology,Tianjin Nankai Hospital,Tianjin 300193,China)

机构地区：[1]天津医科大学南开临床学院,300100 [2]天津市南开医院麻醉科,300193

出　　处：《中华麻醉学杂志》2020年第6期752-755,共4页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金青年科学基金(81601707);国家自然科学基金(81772106)。

摘　　要：目的:评价血红素氧合酶-1(HO-1)在LPS致大鼠肺泡巨噬细胞凋亡中的内源性保护作用及其与内质网应激的关系。方法:大鼠肺泡巨噬细胞NR8383,采用随机数字表法分为4组(n=32):对照组(C组)、LPS组(L组)、Con siRNA组和HO-1 siRNA组。C组常规培养,余3组向培养基加入10μg/ml LPS。Con siRNA组和HO-1 siRNA组于LPS加入前48 h时分别行Con siRNA和HO-1 siRNA转染。LPS处理24 h时采用MTT法测定细胞活力,流式细胞术测定细胞凋亡率,Western blot法检测葡萄糖调节蛋白78(GRP78)、激酶受体样内质网激酶(p-PERK)、增强子结合蛋白同源蛋白(CHOP)、磷酸化的Ⅰ型内质网跨膜蛋白激酶(p-IRE-1)、磷酸化的应激活化蛋白激酶(p-JNK)和半胱氨酸天冬氨酸-12(caspase-12)表达。结果:与C组比较,余3组细胞活力降低,细胞凋亡率升高,HO-1、GRP78、CHOP、p-PERK、p-IRE-1、p-JNK和caspase-12表达上调(P<0.05);与L组比较,HO-1 siRNA组细胞活力降低,细胞凋亡率升高,HO-1表达下调,GRP78、CHOP、p-PERK、p-IRE-1、p-JNK和caspase-12表达上调(P<0.05),Con siRNA组各指标差异无统计学意义(P>0.05);与Con siRNA组比较,HO-1 siRNA组细胞活力降低,细胞凋亡率升高,HO-1表达下调,GRP78、CHOP、p-PERK、p-IRE-1、p-JNK和caspase-12表达上调(P<0.05)。结论:HO-1产生内源性保护作用的机制与抑制内质网应激,减轻LPS诱导大鼠肺泡巨噬细胞凋亡有关。Objective To evaluate the role of heme oxygenase-1(HO-1)-induced endogenous protection in lipopolysaccharide(LPS)-caused apoptosis in rat alveolar macrophages and the relationship with endoplasmic reticulum stress.Methods Alveolar macrophages of rats were randomized into 4 groups(n=32 each)using a random number table method:control group(group C),LPS group(group L),Con siRNA group and HO-1 siRNA group.Cells were cultured normally in group C,and 10μg/ml LPS was added to the culture medium in the other three groups.Con siRNA and HO-1 siRNA transfection was performed at 48 h before adding LPS in Con siRNA and HO-1 siRNA groups.At 24 h of treatment with LPS,MTT method was used to measure the cell viability,flow cytometry was used to determine the cell apoptosis rate,and Western blot was used to detect the expression of glucose regulatory protein 78(GRP78),phosphorylated kinase receptor-like endoplasmic reticulum kinase(p-PERK),CCAAT/enhancer-binding protein homologous protein(CHOP),phosphorylated type I endoplasmic reticulum transmembrane protein kinase(p-IRE-1),phosphorylated stress-activated protein kinase(p-JNK)and caspase-12.Results Compared with group C,the cell viability was significantly decreased,cell apoptosis rate was increased,and the expression of HO-1,GRP78,CHOP,p-PERK,p-IRE-1,p-JNK and caspase-12 was up-regulated in the other three groups(P<0.05).Compared with group L,the cell viability was significantly decreased,cell apoptosis rate was increased,and the expression of HO-1 was down-regulated,and the expression of GRP78,CHOP,p-PERK,p-IRE-1,p-JNK and caspase-12 was up-regulated in group HO-1 siRNA(P<0.05),and no significant change was found in each parameter in group Con siRNA(P>0.05).Compared with group Con siRNA,the cell viability was significantly decreased,cell apoptosis rate was increased,and the expression of HO-1 was down-regulated,and the expression of GRP78,CHOP,p-PERK,p-IRE-1,p-JNK and caspase-12 was up-regulated in group HO-1 siRNA(P<0.05).Conclusion The mechanism of HO-1-induced endogenous p

关 键 词：血红素加氧酶-1 内质网应激 细胞凋亡 脂多糖类 巨噬细胞 肺泡 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]