沙棘总黄酮抗单侧输尿管梗阻大鼠肾脏纤维化作用及机制

Effect of total flavonoids of hippophae on the renal fibrosis of unilateral ureteral obstructive and related mechanism

目的研究沙棘总黄酮对单侧输尿管梗阻(UUO)大鼠肾脏纤维化的影响及相关机制。方法雄性SD大鼠分为假手术组、模型组、依那普利组、沙棘总黄酮低剂量组和沙棘总黄酮高剂量组,UUO手术后连续给药2周,观察肾脏形态变化,测量肾脏皮质厚度、肾脏重量和肾脏长度的变化,Masson三色染色检测沙棘总黄酮对胶原纤维沉积的影响,免疫组化检测沙棘总黄酮对α-平滑肌肌动蛋白(α-SMA)、纤维连接蛋白(Fibronectin)和细胞间隙连接蛋白(Cx43)表达和分布的影响;Western blot检测沙棘总黄酮对α-SMA、钙黏附蛋白-E(E-cadherin)、Fibronectin和转化生长因子-β_(1)(TGF-β_(1))表达的影响;内源性硫化氢(H_(2)S)检测试剂盒检测沙棘总黄酮对肾脏组织和血清H_(2)S水平的影响。结果沙棘总黄酮改善肾脏形态及皮质厚度,抑制胶原纤维的沉积,降低α-SMA、Fibronectin、TGF-β_(1)和Cx43的表达,上调E-cadherin的表达,显著升高肾脏组织和血清中H_(2)S水平。结论沙棘总黄酮可显著改善UUO大鼠肾脏纤维化,抑制上皮细胞-间充质转化形成,减少细胞外基质沉积;沙棘总黄酮抗肾脏纤维化作用可能与其降低TGF-β_(1)表达,升高内源性H_(2)S水平和抑制Cx43表达有关。

Objective To determine the effect of total flavonoids of hippophae(TFH)on the renal fibrosis of unilateral ureteral obstructive(UUO)and related mechanism.Methods Male SD rats were randomly divided into a sham group,a model group,2 TFH groups,and an enalapril group.After the surgery,TFH and enalapril were given via gastric gavage continblously for 2 weeks.Rats in the sham group were given sodium chloride.The changes of renal cortex thickness,weight and length were measured.The interstitial collagen fibrils of the renal tissue were observed with Masson trichrome staining.Theα-SMA,Fibronectin and Cx43 expression and distribution of the renal tissue were observed with immunohistochemistry.The effect of THF on the protein levels ofα-SMA,E-cadherin,Fibronectin and TGF-β_(1) was also measured by Western blot.The renal tissue and the serum H_(2)S were detected by endogenous H_(2)S assay kit.Results TFH ameliorated the renal morphology and cortical thickness,inhibited the collagen fibrils formation,downregulated the expression ofα-SMA,Fibronectin and Cx43,upregulated the expression of E-cadherin in UUO renal tissue,and increased the H_(2)S level in the kindney tissue and the serum.Conclusion TFH can significantly improve renal fibrosis,inhibit epithelial-mesenchymal transformation formation and reduce extracellular matrix deposition in UUO rats.The effect of TFH on the renal fibrosis may be related to the decrease of TGF-β_(1) expression,the increase of endogenous H_(2)S level and the inhibition of Cx43 expression.