摘要
目的观察骨形态发生蛋白-7(bone morphogenetic protein-7,BMP-7)对转化生长因子-β1(transforminggrowth factor-β1,TGF-β1)诱导系膜细胞表达基质金属蛋白酶(matrix metalloprotease,MMP)2、9的影响。方法将体外培养系膜细胞分4组:对照组(NC组)、BMP-7组、TGF-β1组、BMP-7+TGF-β1组(BT组)。酶联免疫吸附测定(en-zyme-linked immunosorbent assay,ELISA)细胞培养上清液MMP2、MMP9、金属蛋白酶组织抑制因子1(tissue inhibitorof metalloproteinase 1,TIMP1)、Ⅳ型胶原(collagen type IV,ColⅣ)、纤维连接蛋白(fibronectin,FN)水平。实时荧光定量反转录聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)测定系膜细胞MMP2、MMP9的表达。Western blot检测MMP2、MMP9蛋白表达水平。结果 NC组MMP2、MMP9、TIMP1、ColⅣ、FN水平及MMP2、MMP9 mRNA和蛋白质的表达与BMP-6组相比无统计学差异。与NC组、BMP-6组相比,TGF-β1组上清液MMP2、MMP9水平显著降低,TIMP1、ColⅣ、FN升高;MMP2、MMP9 mRNA和蛋白质的表达降低。与TGF-β1组相比,BT组MMP2、MMP9水平升高,TIMP1、ColⅣ、FN降低;MMP2、MMP9 mRNA和蛋白质的表达升高。结论 BMP-7可拮抗系膜细胞TGF-β1诱导MMP2、MMP9表达和分泌减少,抑制部分细胞外基质成分的聚积,对肾脏有保护作用。
Objective To investigate the effects of bone morphogenetic protein-7（BMP-7） on the expression of matrix metalloprotease 2（MMP2） and matrix metalloprotease 9（MMP9） induecd by transforming growth factor-β1（TGF-β1） in mesangial cells. Methods The cultured mesangial cells were divided into 4 groups： normal control group（NC group）,BMP-7 group,TGF-β1 group,combined BMP-7 and TGF-β1 treated group（BT group）.The levels of MMP2,MMP9,tissue inhibitor of metalloproteinase 1（TIMP1）,collagen type IV（col IV） and fibronectin（FN） were determined in cultured supernatant by enzyme-linked immunosorbent assay（ELISA）.Real-time quantitative reverse transcription polymerase chain reaction（RT-PCR） and Western blot were used to analyse the assay mRNA and protein expressions of MMP2 and MMP9 respectively. Results The levels of MMP2,MMP9,TIMP1,Col Ⅳ,FN and the mRNA and protein expressions of MMP2 and MMP9 were not significantly different between NC group and BMP-7 group.Compared with NC group and BMP-7 group,the levels and the mRNA and protein expression of MMP2,MMP9 were decreased,and the levels of TIMP1,Col Ⅳ,FN were increased in TGF-β1 group.Compared with TGF-β1 group,the levels and the mRNA and protein expression of MMP2,MMP9 were enhanced,the levels of TIMP1,Col Ⅳ,FN were decreased in BT group. Conclusion BMP-7 can antagonize TGF-β1-induced reduction of MMP2 and MMP9 expression and excretion in mesangial cells,and partly inhibit the accumulation of extracellular matrix to exert the renoprotective effect.
出处
《华南国防医学杂志》
CAS
2011年第5期402-405,共4页
Military Medical Journal of South China
