心肌缺血病人脯氨酸羟化酶2的表达及其干预HL-1缺氧损害的机制  

Expression of Proline Hydroxylase 2 in Patients with Myocardial Ischemia and Its Mechanism of Intervening HL-1 from Hypoxia

作　　者：骆元平 吴钟伟 曾垂旭 LUO Yuanping;WU Zhongwei;ZENG Chuixu(Hainan Western Central Hospital,Danzhou 571700,Hainan,China)

机构地区：[1]海南西部中心医院,海南儋州571700

出　　处：《中西医结合心脑血管病杂志》2022年第12期2168-2173,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：海南省卫生健康行业科研项目(No.19A200078)。

摘　　要：目的探讨脯氨酸羟化酶2(PHD2)在心肌缺血病人中的表达及对心肌细胞HL-1缺氧损害的影响和相关机制。方法采集我院健康体检者和心血管科心肌缺血病人血清,采用酶联免疫吸附法(ELISA)检测PHD2水平。体外培养HL-1,将细胞培养在94%N2、5%CO_(2)和1%O_(2)的培养箱中构建缺氧损伤模型。分别采用四唑盐(MTT)、Transwell和流式细胞术检测细胞存活率、迁移和侵袭能力及细胞凋亡证实缺氧损伤。采用实时荧光定量逆转录聚合酶链反应(RT-qPCR)和蛋白免疫印迹法(Western Blot)检测缺氧后PHD2表达;同时观察过表达PHD2对缺氧诱导的HL-1缺氧损伤的影响。采用Western Blot和ELISA分析过表达PHD2对HL-1核转录因子κB(NF-κB)信号通路相关蛋白的影响。结果相较于健康体检者,心肌缺血病人血清PHD2水平降低。相较于常氧,缺氧损伤模型HL-1存活率下降,迁移和侵袭能力下降,细胞凋亡增加,PHD2下调。过表达PHD2可抑制缺氧诱导的细胞损伤,同时抑制NF-κB信号通路相关蛋白表达。结论心肌缺血病人血清PHD2低表达,同时低氧诱导的HL-1中PHD2低表达,过表达的PHD2可能通过抑制NF-κB信号通路抑制因缺氧诱导的HL-1损伤。Objective To investigate the expression of proline hydroxylase 2(PHD2)in patients with myocardial ischemia and its effect on myocardial cell HL-1 hypoxia damage and related mechanism.Methods The serum of normal physical examination people and myocardial ischemia patients in our hospital were collected,and the level of PHD2 was detected by enzyme-linked immunosorbent assay(ELISA).HL-1 was cultured in vitro,and the cells were cultured in an incubator with 94%N2,5%CO_(2) and 1%O_(2) to construct a hypoxic injury model.Hypoxic injury was confirmed by tetrazolium salt(MTT),cell viability and migration were detected by Transwell,cell apoptosis,and invasion ability were detected by flow cytometry.The expression of PHD2 after hypoxia was detected by real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR)and Western Blot.To observe the effect of overexpression of PHD2 on hypoxia-induced HL-1 hypoxic injury.The effects of PHD2 overexpression on HL-1 nuclear factorκB(NF-κB)signaling pathway-related protein were detected by Western Blot and ELISA analysis.Results The level of PHD2 was decreased in patients with myocardial ischemia than that in normal physical examination people.Compared with the normoxia,the hypoxic injury model HL-1 decreased the survival rate,decreased the migration and invasion ability,increased cell apoptosis,and down-regulated PHD2.Overexpression of PHD2 inhibited hypoxia-induced cell damage and inhibited the expression of NF-κB signaling pathway-related proteins.Conclusion The level of PHD2 in patients with myocardial ischemia was low expressed,and it was also low expressed in hypoxia-induced HL-1.Overexpressed PHD2 might inhibit hypoxia-induced HL-1 injury by inhibiting NF-κB signaling pathway.

关 键 词：心肌缺血 脯氨酸羟化酶2 缺氧 实验研究 

分 类 号：R73[医药卫生—肿瘤]