类叶升麻苷调控Rnd3/NF-κB通路对缺氧/复氧诱导心肌细胞损伤的影响  

Effects of acteoside on hypoxia/reoxygenation-induced cardiomyocyte damage by regulating the Rnd3/NF-κB pathway

作　　者：杨洋 李天发[1] 王军[1] 黄珊[1] 凌学斌 YANG Yang;LI Tianfa;WANG Jun;HUANG Shan;LING Xuebin(Dept.of Cardiovascular Medicine,the First Affiliated Hospital of Hainan Medical College,Haikou 570102,China)

机构地区：[1]海南医学院第一附属医院心血管内科,海口570102

出　　处：《中国药房》2022年第5期542-547,共6页China Pharmacy

基　　金：国家自然科学基金资助项目(No.81860075)。

摘　　要：目的探讨类叶升麻苷调控Rho家族GTP酶3(Rnd3)/核因子κB(NF-κB)通路对缺氧/复氧(H/R)诱导心肌细胞损伤的影响。方法本实验将H9c2心肌细胞分为对照组(不给药、不造模)、H/R组(仅造模)、H/R+AS-L组、H/R+AS-M组、H/R+AS-H组(以上3组先分别给予10、30、90μmol/L类叶升麻苷,再造模)、H/R+pcDNA组[先转染pcDNA(空载体),再造模]、H/R+pcDNA-Rnd3组[先转染pcDNA-Rnd3(Rnd3过表达载体)使Rnd3过表达,再造模]、H/R+AS-H+si-NC组[先转染si-NC(阴性对照),然后给予90μmol/L类叶升麻苷,再造模]、H/R+AS-H+si-Rnd3组[先转染si-Rnd3(Rnd3小分子干扰RNA)抑制Rnd3过表达,然后给予90μmol/L类叶升麻苷,再造模]。各组经相应处理后,检测细胞凋亡率和细胞中乳酸脱氢酶(LDH)释放量、丙二醛(MDA)水平、超氧化物歧化酶(SOD)活性、肿瘤坏死因子α(TNF-α)水平、白细胞介素1β(IL-1β)水平、IL-6水平、Rnd3和NF-κB亚基p65(NF-κB p65)mRNA及蛋白的表达、活化胱天蛋白酶3(Cleaved Caspase-3)和Cleaved Caspase-9蛋白表达的变化。结果给予不同浓度类叶升麻苷均可降低H/R诱导H9c2心肌细胞凋亡率、Cleaved Caspase-3和Cleaved Caspase-9蛋白表达水平、NF-κB p65 mRNA及蛋白表达水平、LDH释放量和MDA、TNF-α、IL-1β、IL-6水平,升高SOD活性和Rnd3 mRNA及蛋白表达水平(P<0.05),且呈剂量依赖性。Rnd3过表达可使H/R诱导H9c2心肌细胞凋亡率,NF-κB p65、Cleaved Caspase-3、Cleaved Caspase-9蛋白表达水平,LDH释放量和MDA、TNF-α、IL-1β、IL-6水平降低,使Rnd3蛋白表达水平和SOD活性升高(P<0.05);而抑制Rnd3过表达,可减弱类叶升麻苷对H/R诱导H9c2心肌细胞的凋亡、氧化应激及炎症反应的抑制作用(P<0.05)。结论类叶升麻苷可通过促进Rnd3表达、抑制NF-κB p65表达来调控Rnd3/NF-κB通路,进而抑制心肌细胞凋亡、氧化应激及炎症反应,从而减轻H/R诱导的心肌细胞损伤。OBJECTIVE To explore the effects of acteoside on hypoxia/reoxygenation(H/R)-induced cardiomyocyte damage by regulating Rho family GTPase 3(Rnd3)/nuclear factor κB(NF-κB)pathway. METHODS The H9c2 cardiomyocyte were divided into control group(no administration,no modeling),H/R group(only modeling),H/R+AS-L group,H/R+AS-M group,H/R+AS-H group(10,30,90 μmol/L acteoside for above 3 groups firstly,and then modeling),H/R+pcDNA group [transfecting pcDNA(empty vector) firstly,and then modeling],H/R + pcDNA-Rnd3 group [overexpression of Rnd3 by transfecting pcDNARnd3(Rnd3 overexpression vector)firstly,and then modeling],H/R+AS-H+si-NC group [transfecting si-NC(negative control)firstly,and then giving 90 μmol/L acteoside and modeling],H/R+AS-H+si-Rnd3 group [inhibiting overexpression of Rnd3 by transfecting si-Rnd3(Rnd3 small interfering RNA) firstly, and then giving 90 μmol/L acteoside and modeling]. After corresponding treatment,the apoptotic rate,release of lactate dehydrogenase(LDH),malondialdehyde(MDA)level,the activity of superoxide dismutase(SOD),the level of tumor necrosis factor α(TNF-α),interleukin 1β(IL-1β) and interleukin-6(IL-6),mRNA and protein expression of Rnd3 and NF-κB subunit p65(NF-κB p65),the expression of aspartate proteolytic enzyme 3(Cleaved Caspase-3)protein and Cleaved Caspase-9 protein were detected. RESULTS Different concentrations of acteoside could reduce the apoptotic rate of H/R-induced H9c2 cardiomyocyte,the protein expressions of Cleaved Caspase-3 and Cleaved Caspase-9,mRNA and protein expressions of NF-κB p65,the levels of LDH release and MDA,TNF-α,IL-1β and IL-6,while increase the activity of SOD and mRNA and protein expressions of Rnd3(P<0.05),in a dose-dependent manner. Overexpression of Rnd3 could decrease the apoptotic rate of H9c2 cardiomyocyte,protein expressions of NF-κB p65,Cleaved Caspase-3 and Cleaved Caspase-9,the levels of LDH release,MDA,TNF-α,IL-1β and IL-6,while increase the protein expression of Rnd3 and the activity of SOD(P<0.05). The inhibition overexpr

关 键 词：类叶升麻苷 Rnd3/NF-κB通路 缺氧/复氧 H9C2心肌细胞 细胞凋亡 氧化应激 炎症反应 

分 类 号：R965.1[医药卫生—药理学]