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目的:通过对肾间质纤维化大鼠肾组织α-平滑肌肌动蛋白(α-SMA)、Ⅳ型胶原及基质金属蛋白酶-9(MMP-9)、金属蛋白酶组织抑制因子-1(TIMP-1)基因表达的研究,探讨和络泄浊方对肾间质纤维化大鼠的治疗作用机理。方法:采用单侧输尿管结扎法(UUO)进行肾间质纤维化模型制备,48只W ister大鼠随机分为假手术组、模型组、中药灌胃组。术后第3、10、20、30天每组各处死4只大鼠。HE染色法观察肾脏病理改变,评定各组大鼠肾小管间质损害程度,免疫组化法检测肾脏α-SMA、Ⅳ型胶原蛋白表达,采用RT—PCR法检测肾脏MMP-9、TIMP-1mRNA的表达。结果:α-SMA、Ⅳ型胶原蛋白表达模型组明显增强(P<0.01),中药灌胃组各时间点的表达均少于模型组,假手术组表达无明显异常。模型组肾小管间质损伤指数及TIMP-1mRNA表达明显高于假手术组(P<0.01),并随时间延长表达增强,中药灌胃组肾小管间质损伤指数及TIMP-1mRNA表达亦高于假手术组(P<0.01),但较模型组表达弱(P<0.01)。MMP-9表达在模型组术后明显升高,第10天达到高峰,中药灌胃组在第20、30天时间点高于模型组(P<0.01)。结论:和络泄浊方通过调节α-SMA、Ⅳ型胶原及MMP-9/TIMP-1的基因表达,影响细胞外基质的降解过程,从而减轻肾间质纤维化。
OBJECTIVE: To investigate the effects of α-smooth muscle actin (α-SMA), type IV collagen, matrix metalloproteinase-9 (MMP-9), and tissue inhibitor of metalloproteinase-1 (TIMP-) on renal interstitial fibrosis in rats. 1) Research on gene expression, and explore the therapeutic mechanism of Hexiezhuo Fang on renal interstitial fibrosis in rats. METHODS: Unilateral ureteral ligation (UUO) was used to prepare renal interstitial fibrosis model. 48 Wistar rats were randomly divided into sham-operated group, model group, and intragastric administration group. On the 3rd, 10th, 20th and 30th days after operation, 4 rats were killed in each group. HE staining was used to observe the renal pathological changes. The degree of renal tubulointerstitial damage was assessed in each group. The expression of α-SMA and type IV collagen was detected by immunohistochemistry. The expression of MMP-9 and TIMP-1 mRNA was measured by RT-PCR. expression. RESULTS: The expression of α-SMA and type IV collagen was significantly increased in the model group (P<0.01). The expression of each group in the intragastric administration group was lower than that in the model group. No significant abnormal expression was observed in the sham operation group. In the model group, the tubulointerstitial injury index and TIMP-1 mRNA expression were significantly higher than those in the sham operation group (P<0.01), and the expression was increased with time. The tubulointerstitial injury index and TIMP-1 mRNA expression were also higher in the Chinese medicine group. The sham operation group (P<0.01) was weaker than the model group (P<0.01). The expression of MMP-9 was significantly increased in the model group and peaked on the 10th day. The intragastric administration group was higher than the model group on the 20th and 30th day (P<0.01). Conclusion: Heluo Xiedefang can reduce the degradation of extracellular matrix by regulating the gene expression of α-SMA, type IV collagen and MMP-9/TIMP-1, so as to reduce renal interstitial fibrosis.