补肾活血中药对系膜增生性肾小球肾炎大鼠肾组织基质金属蛋白酶及组织抑制剂表达的影响

The expression of matrix metalloproteinase and tissue inhibitor in mesangial proliferative glomerulonephritis rat renal tissue and the effects of nourishing kidney and activing blood recipe of traditional Chinese medicine

目的:研究补肾活血中药对系膜增生性肾小球肾炎大鼠肾组织基质金属蛋白酶及组织抑制剂表达的影响。方法:取雄性SD大鼠54只,实验设正常对照组、模型组、补肾活血法药物干预组,每组分2周、4周、8周时间点,每个时间点6只大鼠。肾组织切片分别行HE和PAS染色,进行常规病理学检查。免疫组化检测肾组织基质金属蛋白酶(MMP-2)、TIMP-2、MT1-MMP、TGF-β1的表达。流式细胞术检测肾皮质MMP-2、TIMP-2、MT1-MMP和TGF-β1蛋白表达。结果:模型组大鼠随病程进展肾小球逐渐增大,系膜基质增多,肾小球基底膜增厚。免疫组化染色显示模型组较对照组MMP-2、MT1-MMP表达减少,而药物干预组较模型组表达减少程度轻;模型组较对照组TIMP-2、TGF-β1表达增多,而药物干预组较模型组表达增多程度轻。流式细胞仪检测显示,4周和8周模型组MMP-2的相对值较对照组降低,在统计学上差异具有显著性(P<0.05或P<0.01),而各期模型组TIMP-2和TGF-β1的相对值较对照组升高,MT1-MMP的相对值较对照组降低,差异具有显著性(P<0.01)。结论:系膜增生性肾小球肾炎肾组织MMP-2表达降低、活性减弱,TIMP-2表达升高,TGF-β1具有抑制MMP-2表达,诱导TIMP-2表达的作用。补肾活血法药物能上调MMP-2的表达及活性的部分恢复,同时下调TIMP-2的表达,使得胶原合成减少,在延缓肾小球疾病的进展,防治肾小球硬化方面具有确切疗效。

Objective：To investigate the expression of matrix metalloproteinase（MMP） and tissue inhibitor of matrix metalloproteinase（TIMP）in mesangial proliferative glomerulonephritis（MsPGN） rat renal tissue and the effects of nourishing kidney and activing blood recipe of traditional Chinese medicine.Methods：Chose 54 male rats and separated them into control group, MsPGN group and treated group with nourishing kidney and activing blood recipe of traditional Chinese medicine.Chose six rats per group at the time of the second,the fourth and the eighth week.The sections were treated with HE and PAS stains and the routine pathologic study was carried.Immunohistochemical staining was used to check the expression of MMP-2,TIMP-2, membrance type metalloproteinases-l（MT1-MMP） and transforming growth factor- β1（TGF- β1）.Flow Cytometry was used to check the protein expression of MMP-2,TIMP-2,MT1-MMP and TGF- β1 in rat renal tissue.Results：In MsPGN group,following with the disease progressing,the glomerulus grew graduatly,ECM became more and more,the basement membrane of glomerulus was thicker.The immunohistochemical staining showed the expression of MMP-2 and MT1-MMP was lower compared with control group,but in treated group the level was weaker,the expression of TIMP-2 and TGF- β1 was higher compared with control group,but in treated group the level was weaker too. At the fourth and the eighth week, the Flow Cytometry showed that in MsPGN group the FI of MMP-2 was lower compared with control group（P〈0.05 or P〈0.01）,but in every period the FI of TIMP-2 and TGF- β1 was higher, and of MT1-MMP was lower compared with control group（P〈0.01）.Conclusions：In MsPGN renal tissue,the expression of MMP-2 was reduced,its activity was weakened.The expression of TIMP-2 was increased,TGF- β1 could restrain the expression of MMP-2 and induce the expression of TIMP-2; Nourishing kidney and activing blood recipe of traditional Chinese medicine could induce the expression of MMP-2 and partly recover the activity,restrain the expression of TIMP-2 at the same time,decrease the composition of collagen; Nourishing kidney and activing blood recipe of traditional Chinese medicine could resist glomemlar sclerosis and postpone the course of chronic renal failure.