大黄-丹参药对延缓单侧输尿管梗阻大鼠肾纤维化的研究

Research of Rhubarb-Salvia miltiorrhiza inhibiting renal fibrosis in rats with unilateral ureteral obstruction

目的 探讨大黄-丹参药对对单侧输尿管梗阻(UUO)大鼠肾纤维化的保护作用机制。方法 按照体质量将大鼠随机分为6组:正常组、模型组、对照组和低、中、高剂量实验组,每组10只。除正常组外,其余各组采用单侧输尿管结扎法复制UUO大鼠肾纤维化模型。低、中、高剂量实验组分别按每千克1.5,3.0,6.0 g生药灌胃大黄-丹参药对水煎液;对照组按2.5 g·kg^(-1)灌胃尿毒清溶液;正常组和模型组均灌胃给予等体积的纯净水,灌胃体积为10 mL·kg^(-1),每天1次,连续21 d。用马松染色观察肾纤维化程度并评分,用实时荧光定量聚合酶链反应法检测肾组织microRNA-21(miR-21)、磷酸酶与张力蛋白同源基因(PTEN)和蛋白激酶B(Akt)的表达水平。结果 低、高剂量实验组和对照组、模型组、正常组的肾纤维化评分分别为(3.25±0.49),(3.03±0.40),(2.69±0.37),(3.90±0.15)和0分,肾组织的miR-21相对表达量分别为7.21±0.62,3.77±0.56,3.78±0.39,17.61±0.48和1.00±0.00,PTEN mRNA相对表达量分别为0.61±0.07,0.84±0.08,0.52±0.04,0.35±0.08和1.00±0.00,Akt mRNA相对表达量分别为13.61±0.53,11.52±0.48,9.16±0.95,19.47±1.49和1.00±0.00。低、高剂量实验组和对照组、正常组的上述指标与模型组比较,差异均有统计学意义(均P<0.01)。结论 大黄-丹参药对可以显著抑制UUO大鼠肾组织中miR-21/PTEN/Akt信号通路,从而发挥对肾纤维化大鼠的保护作用。

Objective To investigate the protective mechanism of Rhubarb-Salvia miltiorrhizaon on renal fibrosis in rats with unilateral ureteral obstruction(UUO). Methods The rats were randomly divided into 6 groups according to body weight: normal, model, control and experimental-L,-M,-H groups, with 10 rats per group. Except for the normal group, unilateral ureteral ligation was used to replicate the UUO rat renal fibrosis model. Rats in the experimental-L,-M and-H groups were administrated with Rhubarb-Salvia miltiorrhiza decoction(1.5, 3.0, 6.0 g crude drug·kg^(-1)) by gastrogavage. Rats in control group were gavaged with 2.5 g·kg^(-1)Niaoduqing solution. Rats in normal and model groups were given equal volume of pure water by intragastric administration. Gastric volume was 10 mL·kg^(-1), once daily for consecutive 21 days. The degree of renal fibrosis was observed and scored by Masson staining. The expressionlevels of microRNA-21(miR-21), phosphatase and tensin homolog(PTEN) and protein kinase B(Akt) in renaltissues were detected by real-time fluorescence quantitative polymerase chain reaction.Results The scores of renalfibrosis in experimental-L,-H groups and control, model, normal groups were(3. 25 ± 0. 49),(3. 03 ± 0. 40),(2. 69 ± 0. 37),(3. 90 ± 0. 15) and 0 points;the relative expression levels ofmiR-21 in renal tissues were7. 21 ± 0. 62, 3. 77 ± 0. 56, 3. 78 ± 0. 39, 17. 61 ± 0. 48 and 1. 00 ± 0. 00;the relative expression levels ofPTEN mRNA were 0. 61 ± 0. 07, 0. 84 ± 0. 08, 0. 52 ± 0. 04, 0. 35 ± 0. 08 and 1. 00 ± 0. 00;the relative expression levels ofAktmRNA were 13. 61 ± 0. 53, 11. 52 ± 0. 48, 9. 16 ± 0. 95, 19. 47 ± 1. 49 and 1. 00 ± 0. 00, respectively. The aboveindexes in the experimental-L,-H groups and control, normal groups were significantly different from those in themodel group(allP< 0. 01).Conclusion Rhubarb-Salvia miltiorrhiza can significantly inhibit miR-21/PTEN/Aktsignaling pathway in renal tissues of UUO rats, thus exerting protective effect on renal fibrosis rats.