背景:肝纤维化(HF)是不同慢性肝病进展为肝硬化甚至肝细胞癌的重要阶段。肝星状细胞(HSC)的激活在HF的进展中起着至关重要的作用。IFN-γ/Smad7通路可抑制HSCs活化,而TGF-β1/CUGBP1通路可抑制IFN-γ/Smad7通路的转导,促进HSCs的活化。因此,抑制TGF-β1/CUGBP1途径和激活IFN-γ/Smad7途径逆转HSCs活化并抑制HF。加味桃河承气汤(JTCD)源自古代中医著作《伤寒论》中的桃河承气汤。我们在JTCD中发现了几种抗HF成分,包括人参皂苷Rb1等,但JTCD中抗HF的具体机制尚不清楚。
目的:阐明JTCD通过抑制HSC活化逆转HF的具体机制,并建立中医药治疗HF的科学基础。
方法:我们构建了CCl4诱导的小鼠HF模型,并在体外激活了TGF-β1的人肝星状细胞系(LX-2),之后他们用JTCD和相应的抑制剂治疗。我们通过免疫荧光染色检查了上述两种途径中关键分子的表达,Western印迹和RT-PCR。
结果:JTCD减轻了小鼠的肝损伤并降低了血清ALT和AST水平。此外,JTCD通过降低α-SMA的表达减弱CCl4诱导的HF,COL1A1等标志HSCs在小鼠肝组织中活化。此外,JTCD能有效抑制TGF-β1、p-Smad3、p-p38MAPK,p-ATF2和CUGBP1在体内和体外上调IFN-γ的水平,p-STAT1和Smad7。机械上,在体外使用两种途径的抑制剂后,我们发现JTCD通过恢复TGF-β1/CUGBP1和IFN-γ/Smad7通路的平衡来抑制HSC的活化。
结论:我们证明JTCD通过抑制TGF-β1/CUGBP1信号通路和上调IFN-γ/Smad7信号通路抑制HSCs活化并逆转HF。此外,我们已经确定了JTCD干扰两种途径以抑制HSC活化的特定联系。JTCD是临床治疗HF的有效候选药物。
BACKGROUND: Hepatic fibrosis (HF) is an essential stage in the progression of different chronic liver conditions to cirrhosis and even hepatocellular carcinoma. The activation of hepatic stellate cells (HSCs) plays a crucial role in the progression of HF. IFN- γ/Smad7 pathway can inhibit HSCs activation, while TGF-β1/CUGBP1 pathway can inhibit IFN-γ/Smad7 pathway transduction and promote HSCs activation. Thus, inhibiting the TGF-β1/CUGBP1 pathway and activating the IFN-γ/Smad7 pathway reverses HSCs activation and inhibits HF. Jiawei Taohe Chengqi Decoction (JTCD) was derived from the Taohe Chengqi Tang in the ancient Chinese medical text titled \"Treatise on Febrile Diseases\". We found several anti-HF components in JTCD including ginsenoside Rb1 and others, but the specific mechanism of anti-HF in JTCD is not clear.
OBJECTIVE: To elucidate the specific mechanism by which JTCD reverses HF by inhibiting the activation of HSCs, and to establish a scientific foundation for treating HF with Traditional Chinese medicine (TCM).
METHODS: We constructed a CCl4-induced mice HF model in vivo and activated human hepatic stellate cell line (LX-2) with TGF-β1 in vitro, after which they were treated with JTCD and the corresponding inhibitors. We examined the expression of pivotal molecules in the two pathways mentioned above by immunofluorescence staining, Western blotting and RT-PCR.
RESULTS: JTCD attenuated liver injury and reduced serum ALT and AST levels in mice. In addition, JTCD attenuated CCl4-induced HF by decreasing the expression of α-SMA, COL1A1 and other markers of HSCs activation in mice liver tissue. Moreover, JTCD effectively suppressed the levels of TGF-β1, p-Smad3, p-p38MAPK, p-ATF2, and CUGBP1 in vivo and in vitro and upregulated the levels of IFN-γ, p-STAT1, and Smad7. Mechanically, after using the inhibitors of both pathways in vitro, we found that JTCD inhibited the activation of HSCs by restoring the balance of the TGF-β1/CUGBP1 and IFN-γ/Smad7 pathways.
CONCLUSIONS: We demonstrated that JTCD inhibited HSCs activation and reversed HF by inhibiting the TGF-β1/CUGBP1 signalling pathway and upregulating the IFN-γ/Smad7 signalling pathway. Moreover, we have identified specific links where JTCD interferes with both pathways to inhibit HSCs activation. JTCD is an effective candidate for the clinical treatment of HF.