摘要
目的研究丹参酮ⅡA(TanⅡA)是否可以减轻高糖(HG)诱导的足细胞凋亡和氧化应激。方法CCK-8法筛选合适的TanⅡA干预浓度。MPC-5细胞依次分为:NC组(5.5 mmol/L D-葡萄糖)、MA组(5.5 mmol/L D-葡萄糖和22.5 mmol/L甘露醇)、HG组(25 mmol/L D-葡萄糖)、HG+TanⅡA组(25 mmol/L D-葡萄糖和10μmol/L TanⅡA)和HG+TanⅡA+SIRT1抑制剂(Sirtinol)组(25 mmol/L D-葡萄糖、10μmol/L TanⅡA和20μmol/L Sirtinol)。微板法、WST-1法和可见光法测定氧化应激标志物丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性;流式细胞术测定细胞凋亡;Western blot评估凋亡相关蛋白和沉默信息调节蛋白1(SIRT1)/内皮型一氧化氮合酶(eNOS)通路蛋白的表达。结果TanⅡA提高了HG诱导的MPC-5细胞活力(P<0.05)。与MA组相比,HG组细胞凋亡水平升高,细胞中B淋巴细胞瘤-2基因(Bcl-2)、磷酸化SIRT1(p-SIRT1)和磷酸化eNOS(p-eNOS)表达以及SOD和CAT活性降低,Bcl-2相关X蛋白(Bax)和裂解的caspase-3(cleaved caspase-3)表达及细胞中MDA含量增高(P<0.05)。与HG组相比,TanⅡA组细胞凋亡水平降低,细胞中Bcl-2、p-SIRT1和p-eNOS表达以及SOD和CAT活性升高,Bax和cleaved caspase-3表达及MDA含量降低(P<0.05)。SIRT1抑制剂Sirtinol可减轻TanⅡA对HG诱导的足细胞凋亡和氧化应激的抑制作用(P<0.05)。结论TanⅡA可能通过激活SIRT1/eNOS通路减轻HG诱导的足细胞系PMC-5凋亡和氧化应激反应。
Objective To explore potential mechanisms involved in the reduction of podocyte apoptosis and oxidative stress induced by high glucose(HG)by tanshinoneⅡA(TanⅡA).Methods CCK-8 method was used to screen the appropriate interference concentration of TanⅡA.The MPC-5 cells were divided into NC group(5.5 mmol/L D-glucose),MA group(5.5 mmol/L D-glucose and 22.5 mmol/L mannitol),HG group(25 mmol/L D-glucose),HG+TanⅡA group(25 mmol/L D-glucose and 10μmol/L TanⅡA),and HG+TanⅡA+SIRT1 inhibitors(sirtinol)group(25 mmol/L D-glucose,10μmol/L TanⅡA and 20μmol/L sirtinol).Determina-tion of oxidative stress markers malondialdehyde(MDA),superoxide dismutase(SOD)and catalase(CAT)were made by microplate method,WST-1 method and visible light method.Flow cytometry was used to measure cell apoptosis.Western blot was used to evaluate the expression of apoptosis-related proteins and silent information regulator 1(SIRT1)/endothelial nitric oxide synthase(eNOS)pathway proteins.Results TanⅡA improved the viability of MPC-5 cells induced by HG(P<0.05).Compared with the MA group,the level of cell apoptosis in the HG group increased,the expression of B-cell lymphoma-2 gene(Bcl-2),phosphorylated SIRT1(p-SIRT1)and phosphorylated eNOS(p-eNOS)in the cells,and the activity of SOD and of CAT were reduced,the expression of Bcl-2-associated X protein(Bax)and cleaved caspase-3,and the content of MDA in cells were increased(P<0.05).Compared with the HG group,the level of cell apoptosis in the TanⅡA group reduced,the expressions of Bcl-2,p-SIRT1 and p-eNOS in the cells,and the activity of SOD and CAT were increased,the expression of Bax and cleaved caspase-3,and the content of MDA in cells were reduced(P<0.05).The SIRT1 inhibitor sirtinol attenuated the inhibitory effect of TanⅡA on HG-induced podocyte apoptosis and oxidative stress(P<0.05).Conclusions TanⅡA may reduce HG-induced apoptosis and oxidative stress in podocytes with a potential mechanism of activating the SIRT1/eNOS pathway.
作者
张梦瑶
牛姝
蔡静
张立香
赵志刚
ZHANG Mengyao;NIU Shu;CAI Jing;ZHANG Lixiang;ZHAO Zhigang(The Second Department of Endocrinology,Shijiazhuang People's Hospital,Shijiazhuang 050000;Department of Cardiology,the Second Affiliated Hospital of Hebei Medical University,Shijiazhuang 050000,China)
出处
《基础医学与临床》
2023年第2期277-282,共6页
Basic and Clinical Medicine
基金
2019年河北省医学科学研究重点课题(20190161)。
