丹参酮ⅡA配伍DAPT对UUO大鼠肾间质纤维化的影响

Experimental study of tanshinone ⅡA combined with DAPT treatment on renal interstitial fibrosis in UUO rats

目的研究丹参酮ⅡA配伍DAPT对UUO大鼠肾间质纤维化的保护作用。方法将40只雄性SD大鼠随机分为5组,正常组、模型组、丹参酮ⅡA组(25 mg/kg)、γ-分泌酶抑制剂(DAPT)组(12 mg/kg)和丹参酮ⅡA配伍DAPT组(简称丹参酮ⅡA+DAPT组,其中丹参酮ⅡA和DAPT分别为25 mg/kg、12 mg/kg)。造模第2天给予相应药物干预,共干预14 d。观察肾组织病理变化,检测大鼠肾功能指标:N-乙酰-β-D-氨基葡萄糖苷酶(NAG)和尿微量白蛋白(UMA)的含量。免疫组织化学方法检测α-平滑肌肌动蛋白(α-SMA)、Ⅰ型胶原(ColⅠ)的表达。结果 (1)模型组肾脏组织呈肾小管萎缩,小管间质明显增宽,Masson染色可见大量胶原纤维弥漫分布。各干预组HE、Masson评分显著降低(P <0. 05),丹参酮ⅡA+DAPT组效果最为显著。(2)与正常组比较,模型组NAG、微量白蛋白含量均显著升高(P <0. 05);与模型组比较,各干预组微量白蛋白明显降低(P <0. 05),而NAG呈升高趋势。(3)与正常组比较,模型组α-SMA、ColⅠ平均光密度值显著升高(P <0. 05);与模型组比较,各干预组α-SMA、ColⅠ平均光密度值均明显下降(P <0. 05);各干预组比较,丹参酮ⅡA+DAPT组α-SMA平均光密度值明显低于丹参酮ⅡA组及DAPT组(P <0. 05),丹参酮ⅡA、丹参酮ⅡA+DAPT组ColⅠ平均光密度值显著低于DAPT组(P <0. 05)。结论丹参酮ⅡA配伍DAPT对肾小管间质纤维化有保护作用,一定程度上改善肾功能损害,降低α-SMA、ColⅠ的表达,缓解肾脏纤维化进程。

Objective To observe the effects of tanshinone IIA combined with DAPT on renal interstitial fibrosis in rats with unilateral ureteral obstruction （UUO）. Methods Forty male SD rats were randomly divided into five groups：control group,model group,tanshinone IIA （25 mg/kg） group,DAPT （12 mg/kg） group,tanshinone IIA combined DAPT （1∶ 1） group.On the second day after modeling,the corresponding drug intervention was given for 14 days.The pathological changes of renal tissue was observed and the renal function was detected.The expression of α-smooth muscle actin （α-SMA） and type I collagen （ColI） were detected by immunohistochemistry. Results ①The renal tissue in model group showed the atrophy of the renal tubules,and the interstitium of the tubules was widened.A large number of collagen fibers was diffusely distributed by Masson staining.The HE and Masson scores were decreased.② Compared with the normal group,the levels of NAG and UMA in the model group were higher than those in the normal group.Compared with the model group,the urinary microalbumin in each treatment group was lower.③ The results of immunohistochemistry showed that the average optical density of α-SMA and ColI in the model group was higher than those in the normal group.Compared with the model group,the average optical density of α-SMA and ColI in the treatment group was decreased.Compared with the treatment group,the average optical density of α-SMA in the combined treatment group was lower than that in the tanshinone IIA group and the DAPT group and the average optical density in the tanshinone IIA and the combined treatment group was significantly lower than that in DAPT group. Conclusion Tanshinone IIA combined with DAPT have protective effects on renal tubulointerstitial fibrosis via improving renal function and reducing the expression of α-SMA,ColI.