HIF-1α通过TLR4/NF-κB信号通路对心肌缺血-再灌注大鼠心肌损伤的保护机制分析  

Protective mechanism of HIF-1α on myocardial injury in rats with myocardial ischemia-reperfusion by TLR4/NF-κB signaling pathway

作　　者：韦皓[1] 许玉霞[1] 秦莉[1] 赵玉倩[1] 苇菁桦 何云[1] 刘倩[1] WEI Hao;XU Yu-xia;QIN Li(Department of Paediatrics,Affiliated Hospital of North Sichuan Medical College,Nanchong Sichuan 637000,China)

机构地区：[1]川北医学院附属医院儿科,四川南充637000

出　　处：《临床和实验医学杂志》2019年第10期1017-1020,共4页Journal of Clinical and Experimental Medicine

基　　金：南充市市校合作项目(编号:18SXHZ0562)

摘　　要：目的分析缺氧诱导因子1α(HIF-1α)通过TLR4/NF-κB信号通路保护心肌缺血-再灌注(MIRI)大鼠心肌损伤的作用机制。方法将30只SD大鼠随机分为对照组、MIRI组和HIF-MIRI组各10只,HIF-MIRI组于造模前24 h腹腔注射二甲基乙二酰基甘氨酸(DMOG,20μg/g),MIRI组和HIF-MIRI组大鼠采用手术结扎法制备MIRI模型,对照组只打开心包,不结扎。造模成功后,HE染色检测各组大鼠心肌组织损伤;试剂盒检测血清中肌酸激酶同工酶(CK-MB)和乳酸脱氢酶(LDH)水平; ELISA法测定心肌组织中肿瘤坏死因子(TNF)-α、白介素(IL)-1β和IL-6水平;实时荧光定量(RT)-PCR测定HIF-1αmRNA的表达; Western Blot检测HIF-1α、Toll-样受体4(TLR4)、核转录因子(NF-κB)、天冬氨酸特异性半胱氨酸蛋白酶(Caspase)-3的表达。结果与对照组相比,MIRI组和HIF-MIRI组大鼠血清CK-MB和LDH、炎症因子TNF-α、IL-1β和IL-6水平显著升高(P <0. 01),心肌组织HIF-1αmRNA的表达显著上调(P <0. 01),HIF-1α、TLR4、NF-κB和Caspase-3蛋白的表达显著上调(P <0. 01);与MIRI组相比,HIF-MIRI组大鼠血清CK-MB和LDH水平、炎症因子TNF-α、IL-1β和IL-6显著下降(P <0. 01),心肌组织HIF-1αmRNA和蛋白的表达显著上调(P <0. 01),TLR4、NF-κB和Caspase-3蛋白的表达显著下调(P <0. 01),HE染色显示心肌组织的病理变化明显减轻。结论 HIF-1α可能通过抑制TLR4/NF-κB信号通路抑制炎症因子的释放,降低机体炎症因子水平,改善MIRI心肌损伤。Objective To analyze protective mechanism of HIF-1α against myocardial injury in rats with myocardial ischemia-reperfusion (MIRI) through TLR4/NF-κB signaling pathway. Methods 30 SD rats were randomly divided into control group, MIRI group and HIF-MIRI group, 10 rats in each group. HIF-MIRI group was intraperitoneally injected with dimethyloxalyl glycine (DMOG, 20 μg/g) 24 h before modeling. MIRI group and HIF-MIRI group were given surgical ligation to build MIRI models. The control group only opened the pericardium without ligation. After successful modeling, HE staining was used to detect myocardial tissue damage in each group. Serum CK-MB and LDH were detected by kit. Levels of TNF-α, IL-1β and IL-6 in myocardial tissues were determined by ELISA. HIF-1α mRNA expression was detected by RT-PCR assay. HIF-1α, TLR4, NF-κB and caspase-3 expressions was detected by Western blot. Results Compared with control group, serum CK-MB, LDH, TNF-α, IL-1β and IL-6 significantly increased in MIRI group and HIF-MIRI group ( P <0.01);HIF-1α mRNA expression in myocardial tissues was significantly up-regulated ( P <0.01);HIF-1α, TLR4, NF-κB and caspase-3 protein levels were significantly up-regulated ( P <0.01). Compared with MIRI group, serum CK-MB, LDH, TNF-α, IL-1β and IL-6 decreased significantly in HIF-MIRI group ( P <0.01);HIF-1α mRNA and protein in myocardial tissues significantly increased ( P <0.01);TLR4, NF-κB and caspase-3 protein expressions significantly decreased ( P <0.01). HE staining showed significantly relieved pathological changes in myocardial tissue. Conclusion HIF-1α may inhibit inflammatory factors and relieve MIRI myocardial injury through TLR4/NF-κB signaling pathway.

关 键 词：大鼠 心肌缺血-再灌注 心肌损伤 缺氧诱导因子1Α TLR4/NF-κB 

分 类 号：R54[医药卫生—心血管疾病]