山姜素对缺氧/复氧诱导的心肌细胞损伤的保护作用机制  

Protective mechanism of alpinetin on hypoxia/reoxygenation-induced cardiomyocyte injury

作　　者：陈颖妹 翟任群[1] 周菲[2] CHEN Yingmei;ZHAI Renqun;ZHOU Fei(ECG Room,Hainan General Hospital,Hainan Hospital Affiliated to Hainan Medical College,Haikou 570311,China)

机构地区：[1]海南省人民医院,海南医学院附属海南医院心电图室,海口570311 [2]海南省人民医院,海南医学院附属海南医院心内科,海口570311

出　　处：《中国免疫学杂志》2022年第18期2220-2225,共6页Chinese Journal of Immunology

摘　　要：目的:探讨山姜素对缺氧/复氧(H/R)诱导的心肌细胞损伤的保护作用机制。方法:H/R诱导大鼠心肌细胞H9C2建立细胞损伤模型,不同浓度山姜素处理H9C2细胞,pcDNA、pcDNA-circPRKCI转染H9C2细胞24 h后再进行H/R处理;si-NC、si-circPRKCI分别转染H9C2细胞24 h后置于山姜素培养液中继续培养24 h,再进行H/R处理;试剂盒检测LDH、MDA、SOD水平;ELISA检测MPO、IL-1β、TNF-α水平;MTT、流式细胞术分别检测细胞增殖及凋亡率;化学发光法检测ATP含量;qRTPCR检测circPRKCI、miR-29b-3p表达;双荧光素酶报告基因实验检测circPRKCI与miR-29b-3p的靶向关系;Western blot检测Bcl-2、Bax蛋白表达。结果:山姜素可降低H/R诱导的H9C2细胞LDH、MDA、MPO、IL-1β、TNF-α水平、凋亡率和Bax蛋白水平(P<0.05),而增强SOD活性和提高细胞存活率、ATP含量及Bcl-2蛋白水平(P<0.05),提高circPRKCI表达(P<0.05),降低miR-29b-3p表达(P<0.05),且呈剂量依赖性。H/R诱导的H9C2细胞转染pcDNA-circPRKCI后,LDH、MDA、MPO、IL-1β、TNF-α水平、凋亡率和Bax蛋白水平降低(P<0.05),SOD活性、Bcl-2蛋白水平、细胞存活率和ATP含量升高(P<0.05);circPRKCI可靶向调控miR-29b-3p表达;干扰circPRKCI表达部分逆转山姜素对H/R诱导的H9C2细胞氧化应激、炎症反应、增殖、凋亡及ATP含量的作用。结论:山姜素可通过调控circPRKCI/miR-29b-3p轴抑制细胞氧化应激、炎症反应、凋亡及促进细胞增殖进而减轻H/R诱导的心肌细胞损伤。Objective:To explore protective mechanism of alpinetin on hypoxia/reoxygenation(H/R)-induced cardiomyocyte damage.Methods:H/R was used to induce rat cardiomyocyte H9C2 to establish a cell injury model,and H9C2 cells were treated with different concentrations of alpinetin. pcDNA and pcDNA-circPRKCI were transfected into H9C2 cells for 24 h and did H/R treatment again. si-NC and si-circPRKCI were respectively transfected into H9C2 cells for 24 h and then placed in alpinetin medium for continuous culture for 24 h and did H/R treatment again. Levels of LDH,MDA and SOD were tested by kits. ELISA was used to detect levels of MPO,IL-1β and TNF-α. MTT and flow cytometry were used to detect cell proliferation and apoptosis rates. Chemiluminescence was used to detect ATP content. qRT-PCR was used to detect expressions of circPRKCI and miR-29b-3p. Dual luciferase reporter gene experiment was used to detect targeting relationship between circPRKCI and miR-29b-3p. Western blot was used to detect expressions of Bcl-2 and Bax proteins.Results:Alpinetin could decrease levels of LDH,MDA,MPO,IL-1β,TNF-α,apoptosis rate and protein level of Bax in H9C2 cells induced by H/R(P<0.05),enhance activity of SOD and improve cell survival rate,ATP content and protein level of Bcl-2(P<0.05),increase expression of circPRKCI(P<0.05),while decrease expression of miR-29b-3p(P<0.05),which were in a dose-dependent manner. After H/R-induced H9C2 cells were transfected with pcDNA-circPRKCI,levels of LDH,MDA,MPO,IL-1β,TNF-α,apoptosis rate and protein level of Bax were decreased(P<0.05),while activity of SOD,protein level of Bcl-2,cell survival rate and ATP content were increased(P<0.05). circPRKCI could targeting regulate miR-29b-3p expression. Interfering with circPRKCI expression partially reversed effect of alpinetin on H/R-induced H9C2 cell oxidative stress,inflammation,proliferation,apoptosis and ATP content.Conclusion:Alpinetin can inhibit cell oxidative stress,inflammatory response,apoptosis and promote cell proliferation by regulating ci

关 键 词：山姜素 缺氧/复氧 circPRKCI miR-29b-3p 心肌细胞H9C2 

分 类 号：R361.3[医药卫生—病理学]