三叶香茶菜介导IκB-α磷酸化对CCl_(4)致大鼠肝纤维化的影响

Isodon ternifolius(D.Don)Kudo alleviates carbon tetrachloride-induced liver fibrosis in rats by downregulating IκB-αphosphorylation

目的探讨三叶香茶菜通过影响IκB-α及其磷酸化下调核转录因子(nuclear transcription factor-κB,NF-κB)信号通路减轻四氯化碳诱导大鼠肝纤维化。方法采用CCl_(4)肝纤维化大鼠模型,生化检测血清中谷丙转氨酶(ALT)、羟脯氨酸(HYP),ELISA检测转化生长因子β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)水平、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α),荧光定量PCR法检测大鼠肝组织Toll样受体4(TLR_(4))mRNA、NF-κB p65 mRNA、NF-κB抑制蛋白-α(IκB-α)mRNA的表达,Western blot法检测大鼠肝组织中TLR_(4)、NF-κB p65、p-IκB-α蛋白表达,HE染色评价肝组织病理损害。结果与模型组相比较,三叶香茶菜能够显著抑制肝纤维化大鼠中ALT、HYP、TGF-β1、α-SMA、IL-6、TNF-α水平,改善大鼠肝组织病理损害,显著抑制肝纤维化大鼠TLR_(4)mRNA、NF-κB p65 mRNA的表达,上调IκB-αmRNA的表达,下调TLR_(4)、NF-κB p65蛋白表达,下调p-IκB-α蛋白的表达。结论三叶香茶菜下调IκB-α磷酸化,抑制NF-κB信号通路的活化,达到抗肝慢性炎症损伤作用。

Objective To investigate whether the Isodon ternifolius(D.Don)Kudo attenuates carbon tetrachloride(CCl_(4))-induced liver fibrosis in rats by downregulating the nuclear transcription factor-κB(NF-κB)signaling pathway through interference with IκB-αphosphorylation.Methods Hepatic fibrosis was induced in rats with CCl_(4).Biochemical analysis was used to detect alanine aminotransferase(ALT)and hydroxyproline(HYP),and the enzymelinked immunosorbent assay was used to detect transforming growth factor(TGF-β1),α-smooth muscle actin(α-SMA),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)levels in sera.Fluorescence quantitative PCR was used to detect Toll-like receptor 4(TLR_(4)),NF-κB p65,IκB-αmRNA expression,and Western blot were used to analyze TLR_(4),NF-κB p65,and phosphorylated(p)-IκB-αprotein expression in rat liver tissues.Hematoxylin-eosin staining was used to evaluate histopathological damage in the rat livers.Results Compared with that in the model group,the pathological damage to the livers of rats treated with Isodon ternifolius(D.Don)Kudo extracts was ameliorated;the levels of ALT,HYP,TGF-β1,α-SMA,IL-6 and TNF-αin the serum of rats with liver fibrosis treated with Isodon ternifolius(D.Don)Kudo extracts were significantly reduced;the expression of TLR_(4)and NF-κB p65 mRNA were significantly inhibited;IκB-αmRNA was significantly upregulated;and the expression of TLR_(4),NF-κB p65 and p-IκB-αprotein was downregulated.Conclusions Isodon ternifolius(D.Don)Kudon alleviated liver fibrosis by downregulating IκB-αphosphorylation and inhibiting activation of the NF-κB signaling pathway.