摘要
目的:探究三草尿毒灵抑制肾脏纤维化的作用与机制。方法:选取24只雄性BALB/c小鼠,随机分为假手术组、单侧肾缺血再灌注组、三草尿毒灵组以及环巴胺组,每组6只。制备左侧单侧肾缺血再灌注损伤模型后第3天起,三草尿毒灵组每天给予三草尿毒灵汤剂(9 g/kg)灌胃,环巴胺组每天给予Shh/Gli信号小分子抑制剂环巴胺(5 mg/kg)腹腔注射。术后第10天切除小鼠右侧肾脏,第11天处死小鼠,获取小鼠血清和肾脏进行肾功能检测、免疫组化、Western Blot和qRT-PCR分析。结果:与假手术组相比,单侧肾缺血再灌注组血肌酐和尿素氮水平明显升高(P<0.05);与单侧肾缺血再灌注组比较,三草尿毒灵组、环巴胺组血肌酐和尿素氮水平明显降低(P<0.05);三草尿毒灵组与环巴胺组血肌酐和尿素氮水平比较无显著差异(P>0.05)。单侧肾缺血再灌注组、三草尿毒灵组、环巴胺组肾脏间质纤维化面积均大于假手术组(P<0.05);三草尿毒灵组、环巴胺组肾脏间质纤维化面积均小于单侧肾缺血再灌注组(P<0.05);三草尿毒灵组与环巴胺组肾脏间质纤维化面积比较无显著差异(P>0.05)。单侧肾缺血再灌注小鼠的肾组织中,无论是mRNA水平还是蛋白水平,FN、COL-Ⅲ和α-SMA表达水平均较假手术组小鼠显著升高(P<0.05);三草尿毒灵组和环巴胺组mRNA、蛋白水平,FN、COL-Ⅲ和α-SMA表达水平均低于单侧肾缺血再灌注组(P<0.05);三草尿毒灵组、环巴胺组mRNA、蛋白及FN、COL-Ⅲ、α-SMA表达水平比较无显著差异(P>0.05)。qRT-PCR检测结果显示,单侧肾缺血再灌注组肾组织Gli 1 mRNA水平较假手术组明显升高(P<0.05);三草尿毒灵组和环巴胺组Gli 1 mRNA水平显著低于单侧肾缺血再灌注组(P<0.05);同样,Western blot检测显示三草尿毒灵组和环巴胺组Gli 1蛋白表达低于单侧肾缺血再灌注组(P<0.05)。三草尿毒灵组和环巴胺组Gli 1蛋白表达水平比较无显著差异(P>0.05)。结论:三草尿毒灵可通过调控Shh/Gli信号通路的表达,减轻肾脏纤维化。
Objective:To explore the effect and mechanism of Sancaoniaoduling in alleviating renal fibrosis.Methods:24 Male BALB/c mice were randomly divided into Sham Group,Unilateral Renal Ischemia/Reperfusion Injury Group,Sancaoniaoduling Group and CPN Group,each group with 6 mice.From the 3rd day after the preparation of the left unilateral renal ischema/reperfusion injury model,mice were subjected to daily intragastric administration of Sancaoniaoduling at 9 g/kg in Sancaoniaoduling Group,while the CPN Groups'mice were given daily intraperitoneal injections of CPN at 5 mg/kg.From the 10th day after surgery,mice were subjected to right kidney nephrectomy.Mice were sacrificed on the 11th day after surgery,serum and kidney tissues were collected for further investigation,such as renal function assay,immunohistochemistry study,western blot and qRT-PCR analysis.Results:Compared with Sham Group,serum creatine and blood urea nitrogen level were significantly increased in the UIRI model;compared with UIRI Group,Sancaoniaoduling Group and CPN Group were significantly lower(P<0.05);there was no significant difference between Sancaoniaoduling Group and CPN Group(P>0.05).The area of renal interstitial fibrosis in UIRI Group,Sancaoniaoduling Group,and CPN Group was larger than that of Sham Group(P<0.05);Sancaoniaoduling Group and CPN Group were smaller than UIRI Group(P<0.05);there was no difference between Sancaoniaoduling Group and CPN Group(P>0.05).In the kidney tissue of UIRI mice,regardless of the mRNA level or the protein level,the expression levels of FN,COL-Ⅲandα-SMA were significantly higher than those in the Sham Group(P<0.05);Sancaoniaoduling Group and CPN Group were lower than the UIRI Group(P<0.05);but there was no significant difference between Sancaoniaoduling Group and CPN Group(P>0.05).The results of qRT-PCR showed that the level of Gli 1 mRNA in kidney tissue of UIRI Group was significantly higher than that of Sham Group(P<0.05),and that of Sancaoniaoduling Group and CPN Group were significantly lower than that of UIRI Group(P<0.05).Western blot showed that the Gli 1 protein expression of Sancaoniaoduling Group and CPN Group was lower than that of UIRI Group(P<0.05).But,there was no significant difference in Gli 1 protein expression between Sancaoniaoduling Group and CPN Group(P>0.05).Conclusion:Sancaoniaoduling may inhibits renal fibrosis by regulating Shh/Gli signaling pathway.
作者
周燕妮
苏雨田
许正锦
ZHOU Yan-ni;SU Yu-tian;XU Zheng-jin(The Xiamen Hospital Affiliated to Beijing University of Chinese Medicine,Xiamen,Fujian361000)
出处
《实用中西医结合临床》
2020年第18期1-4,103,共5页
Practical Clinical Journal of Integrated Traditional Chinese and Western Medicine
基金
福建省自然科学基金资助项目(编号:2020D032)。
