刺梨冻干粉对肾纤维化模型大鼠的影响及干预机制研究

Effect of Cili freeze-dried powder on renal fibrosis model in rats and intervention mechanism

目的通过观察刺梨冻干粉对单侧输尿管梗阻(unilateral ureteral obstruction,UUO)模型大鼠肾纤维化及免疫炎症因子的影响,探讨刺梨冻干粉延缓肾纤维化的干预机制。方法将SD雄性大鼠分为4组:假手术组、UUO模型组、氯沙坦钾组和刺梨冻干粉组,前两组均予以蒸馏水,氯沙坦钾、刺梨冻干粉组分别给予氯沙坦钾片(1 mg/100 g)及刺梨冻干粉(300 mg/100 g)灌胃治疗,首次给药后14 d处死大鼠,采集标本,观察各组大鼠肾组织病理改变,应用免疫组织化学法检测α平滑肌肌动蛋白(alpha smooth muscle actinα-SMA)、Ⅲ型胶原(collagenⅢ,Col-Ⅲ)及蛋白免疫印迹法(Western blot)检测肾脏组织转化生长因子β1(transforming growth factor-β1,TGF-β1)、核转录因子-κB p65(NF-κB p65)、Toll样受体2(Toll-like receptor,TLR2)蛋白的表达。结果光镜下,假手术组肾组织结构基本正常,无明显炎性细胞浸润,间质无或少许胶原纤维;模型组肾组织结构紊乱,大量炎性细胞浸润,肾间质可见明显的胶原纤维沉积,氯沙坦钾、刺梨冻干粉组与模型组比较病理改变明显减轻。假手术组大鼠肾组织中仅少量α-SMA、Col-Ⅲ、TGF-β1、NF-κB p65及TLR2蛋白表达,模型组中上述蛋白表达显著升高;与模型组比较,刺梨冻干粉组、氯沙坦钾组大鼠肾组织中α-SMA、Col-Ⅲ、TGF-β1、NF-κB p65、TLR2蛋白显著降低(P<0.01);刺梨冻干粉组肾组织中TLR2、NF-κB p65蛋白较氯沙坦钾组表达显著降低(P<0.05),α-SMA、Col-Ⅲ、TGF-β1蛋白表达二者比较差异无统计学意义(P>0.05)。刺梨冻干粉能够明显减轻UUO大鼠肾功能的损害及输尿管梗阻导致的肾纤维化。结论刺梨冻干粉对UUO模型大鼠肾组织局部的免疫微环境有调节作用,能改善肾纤维化。

Objective To observe the effects of Cili freeze-dried powder on renal fibrosis and inflammatory factors in unilateral ureteral obstruction（ UUO） model rats,and explore the intervention mechanism of Cili freeze-dried powder delaying the process of renal fibrosis. Methods The SD male rats were divided into four groups： sham operated group,model group,losartan potassium group,Cili freeze-dried powder group. The former two groups were given normal distilled water,and losartan potassium group and Cili freeze-dried powder group were treated with losartan potassium（ 1 mg/100 g） and Cili freeze-dried powder（ 300 mg/100 g） respectively. Model rats were sacrificed at 14 th day after first administration. The specimens were collected,and the pathological changes of renal tissue were observed. Immunohistochemical method was used to detect alpha-smooth muscle actin（ α-SMA）,collagen type Ⅲ（ Col-Ⅲ）,and Western blot was used to detect protein expression level of TGF-β1,NF-κB p65 and TLR2. Results Under light microscope,the renal tissue structure in the sham operated group was basically normal and there was no infiltration of inflammatory cells in the stroma,without Collagennous fiber or a little Collagennous fiber changes. The renal tissue structure in the model group was disor-dered,and a large number of inflammatory cell infiltration and obvious interstitial collagen deposits were seen in the interstitium of the kidney. As compared with model group,pathological changes were significantly alleviated in losartan potassium group and Cili freeze-dried powder group,and only a small amount of expression of α-SMA,Col-Ⅲ,TGF-β1,NF-κB p65 and TLR2 was detected in renal tissue in sham operated group. In the model group,the expression of these proteins increased significantly. As compared with the model group,α-SMA,Col-Ⅲ,TGF-β1,NF-κB p65 and TLR2 proteins were significantly down-regulated in the losartan potassium group and the Cili freeze-dried powder group（ P 0. 01）. As compared with the Losartan potassium group,the expression of TLR2 and NF-κB p65 proteins was significantly reduced in the Cili freeze-dried powder group（ P 0. 05）,and there was no statistically significant difference in the expression of α-SMA,Col-Ⅲ and TGF-β1（ P 0. 05）. Conclusions The Cili freeze-dried powder on immune renal tissue in the rat UUO model can modulate the local microenvironment,and can improve the renal fibrosis.