(1)Department of Biochemistry and Molecular Biology,Harbin Medical University, Harbin 150081, China;2)Department of Experimental Research Center,the Second Affiliated Hospital of Harbin Medical University,Harbin 150086,China; 3)Biotechnology Experimental Center for Teaching,Harbin Medical University, Harbin 150081, China)
Abstract:Thioredoxin1(Trx1)is an important thiol disulfide oxidoreductase in the cell,which plays a crucial role in the maintenance of cellular redox homeostasis against oxidative stress.To study the effects of Trx1 overexpression on matrix metalloproteinase 9(MMP9) expression,HBZY-1 cells were transfected with either PIRESpuro3-sense-Trx1 and PIRESpuro3-antisense-Trx1.The Trx1 expression was assayed by RT-PCR and Western blotting.The mRNA level of MMP9 and its enzyme activity were determined by RT-PCR and gelatin zymography.The ROS level was detected by flow cytometry.The results showed that the MMP9 mRNA and enzyme activity were significantly increased at 12,24 and 48 hours under high glucose condition (P<0.05).A similar response was showed in antisense-Trx1 transfected cells (P<0.01),but for sense-Trx1 transfected cells,no such increase was observed (P>0.05).The ROS level under high glucose was significantly higher than that under normal glucose at 12,24 and 48 hours (P<0.01).In sense-Trx1 transfected cells,the ROS level was significantly lower than that in antisenseTrx1 transfected cells (P<0.05).Taken together,the inhibition of MMP9 expression and activity by Trx1 over-expression was regulated by the ROS level under high glucose conditions in HBZY-1 cells.This suggests that Trx1 may be applied for the treatment of diabetic nephropathy.
房绍红,,刘心平,靳玉宏,贾慧婕,那丽颖,郑海霞,周宏博,. 高糖环境下Trx1过表达对HBZY-1细胞中MMP9表达的影响[J]. 中国生物化学与分子生物学报, 2009, 25(05): 465-472.
FANGShaoHo-ng,,LIUXin-Ping,JINYu-Hong,JIAHui-Jie,NALi-Ying,ZHENGHai-Xia,ZHOUHong-Bo,. Effect of Trx1 Overexpression on Expression Level of MMP9 under High Glucose Condition in HBZY-1. Chinese Journal of Biochemistry and Molecular Biol, 2009, 25(05): 465-472.