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[摘要]
目的研究肾康注射液对慢性马兜铃酸肾病(CAAN)模型大鼠肾间质纤维化的拮抗效应。方法雄性SD大鼠随机分为对照组、模型组和干预组,每组6只。于0、1、4、8、12周末分别检测体质量、尿糖2、4 h尿蛋白定量和肌酐清除率(Ccr);第12周末处死大鼠留取肾组织,行Masson染色观察肾间质纤维化程度;并用逆转录实时定量聚合酶链反应和免疫组织化学方法检测肾组织中转化生长因子-β1(TGF-β1)、结缔组织生长因子(CTGF)、纤溶酶原激活物抑制物-1(PAI-1)、金属蛋白酶组织抑制物-1(TIMP-1)和Ⅰ型胶原(Col I)mRNA及蛋白表达。结果与对照组比较,模型组和干预组大鼠体质量明显减轻(P<0.01),但干预组大鼠体质量明显高于模型组,12周末时存在显著差异(P<0.05)。与对照组相比,模型组大鼠24 h尿蛋白定量显著上升(P<0.01),Ccr显著下降(P<0.01);12周时肾间质纤维化面积显著增加(P<0.01);肾组织内上述各检测指标的mRNA及蛋白表达均显著上调(P<0.01)。与模型组相比,干预组大鼠各时间点Ccr均显著升高(P<0.01);第12周时24 h尿蛋白定量及肾间质纤维化面积均显著降低(P<0.05、0.01);上述高表达的mRNA及蛋白指标均被显著抑制(P<0.05)。结论肾康注射液可能通过抑制肾组织促细胞外基质(ECM)合成因子(TFG-β1、CTGF)及抗ECM降解因子(TIMP-1、PAI-1)生成,而改善CAAN的肾间质纤维化及肾功能。
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[Abstract]
Objective To study the antagonizing effects of Shenkang Injection(SKI) on renal interstitial fibrosis in model rats with chronic aristolochic acid nephropathy(CAAN).Methods Eighteen male SD rats were equally divided into the following three groups: control group,model gruop,and intervention group.Body weight,urinary glucose,24 h urinary protein excretion,and creatinine clearance(Ccr) were measured at the end of 0,1,4,8,and 12 week,respectively.At the end of the 12 week,all the rats were sacrificed and the renal pathological examination of each rat was performed.The mRNA and protein expression of transforming growth factor-β1(TGF-β1),connective tissue growth factor(CTGF),plasminogen activator inhibitor-1(PAI-1),tissue inhibitor of metalloproteinase-1(TIMP-1),and type I collagen(Col I) in kidney tissue were determined by realtime(RT)-PCR and immunohistochemistry staining,respectively.Results Compared with control group,body weight of rats in intervention group and intervention group were significantly decreased(P0.01);body weight of rats in intervention group was higher than that in intervention group,with a significant difference at the end of 12 week(P0.05).Compared with control group,the 24 h urinary protein excretion of rats in model group and intervention group increased significantly(P0.01);but the 24 h urinary protein excretion of rats in intervention group was lower than that in model group(P0.05).Compared with control group,the Ccr of model rats decreased significantly(P0.01);the Ccr of rats in intervention group was significantly higher than that in model group(P0.01).Compared with control group,the relative area of interstitial fibrosis of kidney tissue,in model rats was significantly enlarged(P0.01).The mRNA and protein expression of TGF-β1,CTGF,PAI-1,TIMP-1,Col I in kidney tissue was significantly up-regulated(P0.01) in the model rats.Compared with the control group.The increased mRNA and protein expression of TGF-β,CTGF,PAI-1,TIMP-1,Col I were all significantly down-regulated in intervention group(P0.05).Conclusion SKI could inhibit the production of promoting extracellular matrix(ECM) synthesis factors(TGF-β1 and CTGF) and antagonizing ECM degradation factors(TIMP-1 and PAI-1) in kidney tissue,which might be the mechanism that SKI alleviates the renal interstitial fibrosis and improves the renal function in CAAN rats.
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[基金项目]
卫生部属(管)医疗机构临床学科重点项目