木棉花提取物调控lncRNA TALNEC2表达对缺氧/复氧心肌细胞氧化应激损伤的保护作用  

The Protective Effect of Kapax Extract on Cardiomyocytes Hypoxia/Reoxygenation Induced Oxidative Stress Injury by Regulating lncRNA TALNEC2 Expression

作　　者：陈静[1] 李芹[2] CHEN Jing;LI Qin(Baoji Vocational&Technical College,Baoji 721003,Shaanxi,China)

机构地区：[1]宝鸡职业技术学院,陕西宝鸡721003 [2]宝鸡市人民医院,陕西宝鸡721000

出　　处：《中西医结合心脑血管病杂志》2022年第24期4478-4483,共6页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：2021年度宝鸡市哲学社会科学项目(No.BJSKZX-202113)。

摘　　要：目的 探讨木棉花提取物对缺氧/复氧心肌细胞氧化应激损伤的影响,并进一步探讨其保护机制是否与调控长链非编码RNA(lncRNA)2号染色体表达的肿瘤相关lncRNA(TALNEC2)表达有关。方法 采用细胞计数试剂盒(CCK)-8法测定低剂量、中剂量、高剂量(10 mg/L、40 mg/L、160 mg/L)木棉花提取物对心肌细胞H9c2活性的影响。体外培养H9c2细胞建立缺氧/复氧损伤模型。将H9c2细胞分为对照组、模型组、低剂量+模型组、中剂量+模型组、高剂量+模型组、阴性对照(si-NC)+模型组、小干扰RNA(si-lncRNA) TALNEC2+模型组、空载体质粒(pcDNA)+高剂量+模型组、pcDNA-lncRNA TALNEC2+高剂量+模型组。采用流式细胞术检测细胞凋亡率;采用酶标法检测细胞内超氧化物歧化酶(SOD)活力和丙二醛(MDA)含量,细胞培养上清液中乳酸脱氢酶(LDH)水平;实时定量酶联免疫吸附法(RT-PCR)检测lncRNA TALNEC2表达水平。结果 与对照组比较,模型组H9c2细胞凋亡率、LDH释放量、MDA含量、lncRNA TALNEC2表达水平增加(P<0.05),SOD活力降低(P<0.05)。与模型组比较,低剂量+模型组、中剂量+模型组、高剂量+模型组H9c2细胞凋亡率、LDH释放量、MDA含量、lncRNA TALNEC2表达水平降低(P<0.05),SOD活力升高(P<0.05)。与si-NC+模型组比较,si-lncRNA TALNEC2+模型组H9c2细胞凋亡率、LDH释放量、MDA含量降低(P<0.05),SOD活力升高(P<0.05)。与pcDNA+高剂量+模型组比较,pcDNA-lncRNA TALNEC2+高剂量+模型组H9c2细胞凋亡率、LDH释放量、MDA含量升高(P<0.05),SOD活力降低(P<0.05)。结论 木棉花提取物通过下调lncRNA TALNEC2表达保护心肌细胞免受缺氧/复氧诱导的氧化应激损伤。Objective To investigate the effect of kapok extract on hypoxia/reoxygenation induced cardiomyocytes oxidative stress damage, and to explore whether its protective mechanism is related to regulate long non-coding RNA(lncRNA) tumors tumor associated long noncoding RNA expressed on chromosome 2(TALNEC2).Methods Cell counting kit-8(CCK-8) was used to detect the effects of low, medium and high doses(10 mg/L,40 mg/L,160 mg/L) of kapok extract on the activity of cardiomyocytes H9c2.Cardiomyocytes H9c2 were cultured in vitro to establish the hypoxia/reoxygenation injury model.Cardiomyocytes H9c2 were divided into control group, model group, low dose+model group, medium dose+model group, high dose+model group, si-NC+model group, si-lncRNA TALNEC2+model group, pcDNA+high dose+model group, pcDNA-lncRNA TALNEC2+high dose+model group.Cell apoptosis rate was detected by flow cytometry;The activity of superoxide dismutase(SOD) and the content of malondialdehyde(MDA) in cells and the level of lactate dehydrogenase(LDH) in the supernatant of cell culture were detected by enzyme-labelled immunosorbent assay.The expression level of lncRNA TALNEC2 was detected by real-time quantitative polymerase chain reaction.Results Compared with the control group, the apoptosis rate, LDH release, MDA content and lncRNA TALNEC2 expression level of H9c2 cells in the model group were increased(P<0.05),while SOD activity was decreased(P<0.05).Compared with the model group, the apoptosis rate, LDH release, MDA content, lncRNA TALNEC2 expression level and SOD activity of H9c2 cells in the low dose+model group group, medium dose+model group, and high dose+model group were decreased(P<0.05),while SOD activity was increased(P<0.05).Compared with the si-NC+model group, the apoptosis rate, LDH release, MDA content of H9c2 cells in the si-lncRNA TALNEC2+model group were decreased(P<0.05),while SOD activity was increased(P<0.05).Compared with pcDNA+high-dose+model group, the apoptosis rate, LDH release, MDA content of H9c2 cells in the pcDNA-lncRNA TALNEC2+

关 键 词：心肌细胞 H9C2 木棉花提取物 缺氧/复氧 长链非编码RNA 实验研究 

分 类 号：R285[医药卫生—中药学]