miR-26a-5p基于PHD2/HIF-1α通路对慢性心力衰竭大鼠心肌细胞凋亡作用的研究  

Apoptosis Inhibition Effect of MiR-26a-5p on Cardiac Function in Rats with Chronic Heart Failure by Regulating PHD2/HIF-1α Signaling Pathway

作　　者：赵洁 李嘉[1] 赵小丹 丁鹏 苗昌荣 ZHAO Jie;LI Jia;ZHAO Xiaodan;DING Peng;MIAO Changrong(Internal Medicine-Cardiovascular Department,Tangshan Union Medical College Hospital,Tangshan 063000,China)

机构地区：[1]唐山市协和医院心内科,河北唐山063000

出　　处：《标记免疫分析与临床》2023年第3期492-498,共7页Labeled Immunoassays and Clinical Medicine

基　　金：河北省医学科学研究课题(编号:20201547)。

摘　　要：目的通过缺氧诱导信号通路PHD2/HIF-1α探究miR-26a-5p对慢性心力衰竭(CHF)模型大鼠心肌细胞凋亡的影响。方法将大鼠随机分为假手术组、CHF组、NC组和miR-26a-5p组,每组16只。CHF组、NC组与miR-26a-5p组建立大鼠CHF模型,假手术组除不进行结扎外其他操作相同。NC组和miR-26a-5p组分别尾静脉注射5nmol NC-agomiR和miR-26a-5p-agomiR,每3天1次,连续注射4周,假手术组和CHF组通过尾静脉注射给予等量生理盐水。对大鼠心肌组织进行masson染色,实时定量PCR法检测大鼠心肌组织miR-26a-5p表达水平,Tunel法检测大鼠心肌组织进行凋亡检水平,ELISA法检测心肌组织氧化损伤因子超氧化物歧化酶(SOD)和丙二醛(MDA)水平,实时定量PCR法和Western blotting法检测心肌组织脯氨酸羟化酶2(PHD2)及低氧诱导因子-1α(HIF-1α)mRNA及蛋白表达水平。结果与假手术组相比,CHF组和NC组出现心肌肥大并伴有大量纤维细胞出现,miR-26a-5p组大鼠心肌肥大有所缓解,纤维细胞减少;CHF组和NC组心肌组织miR-26a-5p表达水平有降低趋势,miR-26a-5p组表达显著上升(P<0.01);CHF组和NC组心肌细胞显著增加(P<0.01),miR-26a-5p组心肌细胞凋亡与CHF组和NC组相比显著降低,但仍高于假手术组(P<0.01);CHF组和NC组大鼠心肌组织SOD显著降低、MDA表达显著升高(P<0.01),与CHF组和NC组相比,miR-26a-5p组SOD显著升高、MDA表达显著降低(P<0.01);CHF组和NC组PHD2 mRNA和蛋白均有上调趋势,HIF-1α有下调趋势,与CHF组和NC组相比,miR-26a-5p组PHD2 mRNA及其蛋白表达下调,HIF-1α表达上调(P<0.05)。结论miR-26a-5p可以抑制CHF大鼠心肌细胞凋亡,提高SOD表达,降低MDA表达,降低氧化损伤,其机制与缺氧诱导信号通路PHD2/HIF-1α的调控相关。Objective To explore the effect of miR-26a-5p on cardiomyocyte apoptosis in rats with chronic heart failure though PHD2/HIF-αpathway.Methods 64 SPF Wistar rats were randomly divided into the sham operation group(n=16),CHF group(n=16),NC group(n=16)and miR-26a-5p group(n=16).CHF group,NC group and miR-26a-5p group were used to establish the rat model of chronic heart failure.The sham operation group received the same operation except ligation.NC group and miR-26a-5p group were injected with 5nmol NC agomiR and miR-26a-5p-agomiR through caudal vein respectively,once every 3 days for 4 weeks.The sham operation group and CHF group were injected with the same amount of saline through caudal vein.Masson staining was performed on the myocardium of rats in each group.The expression level of miR-26a-5p was detected by real-time quantitative PCR,while the apoptosis was detected by TUNEL,and the levels of oxidative damage factors superoxide dismutase(SOD)and malondialdehyde(MDA)were detected by ELISA.Real time quantitative PCR and western blot were used to detect proline hydroxylase 2(PHD2)and hypoxia inducible factor-1 in myocardial tissueα(HIF-1α).Results Compared with the sham operation group,myocardial hypertrophy and a large number of fibroblasts appeared in CHF group and NC group,while miR-26a-5p group,myocardial hypertrophy relieved and fibroblasts reduced.The expression of miR-26a-5p decreased in CHF group and NC group,while the expression of miR-26a-5p increased significantly in miR-26a-5p group(P<0.01).The expression of cardiomyocytes in CHF group and NC group increased significantly(P<0.01).Cardiomyocyte apoptosis in miR-26a-5p group was significantly lower than CHF group and NC group(P<0.01).SOD decreased significantly and MDA increased significantly(P<0.01),compared with CHF group and NC group.SOD in miR-26a-5p group increased significantly and MDA decreased significantly(P<0.01).PHD2 mRNA and protein were up-regulated and HIF1-αdown-regulated in CHF group and NC group,compared with CHF group and NC group.P

关 键 词：miR-26a-5p 慢性心力衰竭 凋亡 氧化损伤 PHD2/HIF-1α信号通路 

分 类 号：R541.6[医药卫生—心血管疾病]