扶肾方对UUO大鼠肾脏病理及CTGF、FN、Vimentin表达的影响

Effect of Fushen Decoction on the expression of CTGF,FN and Vimentin proteins in renal tissues of UUO rats

关键词:  扶肾方  肾间质纤维化  结缔组织生长因子  纤连蛋白  波形蛋白

Key words:  Fushen Decoction  renal interstitial fibrosis  CTGF  FN protein  Vimentin protein

摘要:

[目的] 研究扶肾方对单侧输尿管梗阻（UUO）大鼠肾组织结缔组织生长因子（CTGF）、纤连蛋白（FN）、波形蛋白（Vimentin）表达的影响。[方法] 健康雄性SD大鼠50只，适应性饲养1周后，按照随机原则将大鼠分为假手术组（12只）与造模组（38只），造模组在手术后7 d随机选取大鼠2只以验证造模是否成功，余造模组大鼠随机分为模型组（12只）、中药低、高剂量组（各12只）。除假手术组外，其余各组均以单侧输尿管结扎的方法制作肾间质纤维化模型，假手术组仅游离输尿管。各组大鼠在术后7 d开始灌胃给药，所有大鼠均按照每只每天2 mL进行灌胃，假手术组与模型组灌服生理盐水，中药低剂量组和高剂量组分别按照扶肾方溶度2.52和5.04 g/mL进行灌胃。用苏木精-伊红（HE）染色及电镜观察肾脏病理形态变化；蛋白免疫印迹（Western Blot）检测肾组织CTGF、FN、Vimentin的表达。[结果] 光镜下观察，HE染色示假手术组肾小球形态完整，肾小管结构正常，肾间质正常；模型组术后28 d，肾小球数量减少，肾皮质显著变薄，肾小管萎缩、坏死、消失明显，肾间质纤维化程度严重；中药低、高剂量组肾组织病变较模型组轻。电镜示对比假手术组，模型组肾组织病变明显，中药低、高剂量组肾组织病变较模型组轻。Western Blot检测显示，在术后14、28 d时，与假手术组比较，模型组、中药各剂量组CTGF、FN、Vimentin表达明显增多（P<0.05）；中药低、高剂量组与模型组比较，在术后14 d CTGF、FN、Vimentin表达明显下降（P<0.05）；与低剂量组比较，在术后28 d高剂量组CTGF、FN、Vimentin表达明显下降（P<0.05）。[结论] 扶肾方可以降低UUO大鼠CTGF、FN、Vimentin的表达，随着梗阻时间的延长，高剂量效果更好。

Abstract:

[Objective] To investigate the effects of Fushen Decoction on the expression of connective tissue growth factor (CTGF), fibronectin(FN), and vimentin in renal tissue of UUO rats with unilateral ureteral obstruction.[Methods] After 1 week of adaptive feeding, 16 healthy male SD rats were randomly divided into sham operation group, model group, low-dose group and high-dose group with 4 rats in each group. The renal interstitial fibrosis model was made by ligation of unilateral ureter in all groups except the sham operation group, while the ureter was only free in the sham operation group. The 7 days after the operation, the sham operation group and the model group were given normal saline by gavage at the rate of 2 mL/day, while the low-dose group and the high-dose group were given normal saline at the rate of 2.52 g/mL and 5.04 g/mL respectively. Two rats in each group were sacrificed 14 d and 28 d after surgery, respectively. The pathological changes of kidney were observed by HE staining and electron microscopy. Western blot was used to detect the expression of CTGF, FN and Vimentin proteins in renal tissues.[Results] Under the light microscope, HE staining showed that the glomeruli in the sham group were intact, with normal tubules and normal interstitium. The 28 days after the operation, the number of glomeruli in the model group was reduced, the renal cortex was significantly thinner than before, the tubules atrophy, necrosis and disappearance were obvious, and the degree of renal interstitial fibrosis was further aggravated. The pathological changes of renal tissue in the low and high dose groups were lighter than those in the model group. Electron microscopy showed that compared with the sham operation group, the renal tissue lesions in the model group were obvious, and those in the low and high dose TCM groups were less severe than those in the model group. Western blot showed that the protein expressions of CTGF, FN and Vimentin were significantly increased in the model group, the low-dose group and the high-dose group compared with the sham group (P<0.05). Compared with the model group, CTGF, FN and Vimentin protein expression were significantly decreased 14 days after operation (P<0.05). Compared with the low-dose group, the protein expressions of CTGF, FN and Vimentin were significantly decreased in the high-dose group 28 days after the operation (P<0.05).[Conclusion] Fushen Decoction could reduce the expression of CTGF, FN and Vimentin in UUO rats, and the effect was better with the prolongation of obstruction time.