葛根素对高糖诱导HUVEC-12细胞氧化损伤的保护作用

Protective effects of puerarin on high glucose-induced oxidative injury in HUVEC-12 cells

目的研究葛根素对高糖诱导人脐静脉内皮细胞HUVEC-12氧化损伤的保护作用。方法用含100 mmoL/L葡萄糖和10、25、50μmo L/L葛根素的培养基共同孵育HUVEC-12细胞36 h后,测定细胞存活率,细胞培养液中乳酸脱氢酶(LDH)水平及半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)活性,细胞内活性氧(ROS)水平及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)活性及丙二醛(MDA)、还原型谷胱甘肽(GSH)水平。qPCR检测细胞中SIRT1、PGC-1α mRNA表达,酶联免疫吸附法检测培养液中SIRT1、PGC-1α蛋白含有量。结果葛根素可增加HUVEC-12细胞存活率,抑制细胞LDH释放,清除ROS,降低MDA含有量与Caspase-3活性,增加SOD、CAT活性和GSH含有量。同时,葛根素还能提高SIRT1、PGC-1α mRNA表达及蛋白含有量。结论葛根素能通过激活SIRT1/PGC-1α通路来保护高糖诱导HUVEC-12细胞氧化损伤。

AIM To explore the action mechanism of puerarin＇s protective effects against oxidative stress of HUVEC-12 cells induced by high glucose. METHODS HUVEC-12 cells cultured with 100 mmol/L glucose medium and 10,25,50 μmoL/L puerarin for 36 h had the cell proliferation,the levels of lactate dehydrogenase（ LDH）,intracellular reactive oxygen species（ ROS）,the activities of caspase-3,superoxide dismutase（ SOD）and catalase（ CAT）,and the contents of malondialdehyde（ MDA） and glutathione（ GSH） measured. The mRNA expressions of SIRT1 and PGC-1α were detected by real-time fluorescence quantitative PCR,and the protein contents of SIRT1 and PGC-1α were determined by enzyme-linked immunosorbent assay. RESULTS The puerarin treatment to HUVEC-12 cells resulted in markedly lowered LDH level,caspase-3 activity,intracellular levels of MDA and ROS,and notable improvement of the cell viability,the activities of SOD and CAT,GSH content,the mRNA expressions and the protein contents of SIRT1 and PGC-1α as well. CONCLUSION The protective effect of puerarin on high glucose-induced oxidative damage of HUVEC-12 cells may be attributed to the SIRT1/PGC-1α pathway activation.