抗纤灵冲剂对肾间质纤维化大鼠结缔组织生长因子蛋白表达的影响

Effects of Kangxianling Granules on expression of connective tissue growth factor protein in rats with renal interstitial fibrosis

目的:从结缔组织生长因子(CTGF)蛋白表达的角度,研究抗纤灵冲剂干预肾间质纤维化的作用机制。方法:将96只SPF级SD大鼠随机分为假手术组24只,造模组72只,造模组大鼠应用UUO法制备肾间质纤维化大鼠模型,造模成功后分为模型组、抗纤灵组、氯沙坦组。药物干预后分批处死,留取尿液、左侧肾组织。应用免疫组化法测定CTGF在肾间质细胞及肾小管基底膜的蛋白表达;用放射免疫法测定尿β2-MG水平。结果:与假手术组比较,模型组大鼠CTGF蛋白表达和尿β2-MG水平显著增加(P<0.01,P<0.05);与模型组比较,抗纤灵组和氯沙坦组各时间点CTGF蛋白表达显著减少(P<0.05,P<0.01),抗纤灵组各时间点尿β2-MG水平显著降低(P<0.01,P<0.05);氯沙坦组尿β2-MG水平7d时显著降低(P<0.01);抗纤灵组各时间点CTGF蛋白表达显著低于氯沙坦组(P<0.01);抗纤灵组14d及28d时尿β2-MG水平显著低于氯沙坦组(P<0.01)。结论:抗纤灵冲剂通过调控CTGF基因表达,下调CTGF在肾间质细胞及肾小管基底膜的蛋白表达,从而达到改善肾间质纤维化、保护肾小管功能的作用。

Objective:To study the mechanism of Kangxianling Granules in intervention renal interstitial fibrosis from the expression of connective tissue growth factor(CTGF)protein.Methods:A total of 96 SPF Sprague-Dawley rats were randomly divided into sham-operated group(n=24)and model group(n=72).The model group was prepared by UUO method,they were randomly divided into model group,Kangxianling(KXL)group and Losartan group after successful modeling.After drug intervention,they were killed in batches,the urine and left renal tissue were collected.The protein expression of CTGF in renal stromal cells and basement membrane of renal tubules was determined by immunohistochemistry,and urine β2-MG levels were determined by radioimmunoassay.Results:Compared with the sham-operated group,the expression of CTGF protein and the level of β2-MG in the model group were significantly increased(P<0.01,P<0.05).Compared with the model group,the expression of CTGF protein in the KXL group and losartan group were significantly reduced at each time point(P<0.05,P<0.01).The level of β2-MG was significantly reduced in KXL group at each time point(P<0.01,P<0.05).The level ofβ2-MG in losartan group decreased significantly at 7d(P<0.01).The expression of CTGF protein in KXL group at each time point was significantly lower than that in losartan group(P<0.01).At 14d and 28d,the level ofβ2-MG in KXL group was significantly lower than that in losartan group(P<0.01).Conclusion:Kangxianling Granules can improve renal interstitial fibrosis and protect renal tubular function by regulated the expression of CTGF gene and down-regulated the protein expression of CTGF in renal stromal cells and basement membrane of renal tubules.