Li Qian, Yan Shu-Guang, Hui Yi, Li Jing-Tao, Wei Hai-Liang, Zhang Hong
Medical Experiment Center, Shaanxi University of Chinese Medicine, Xianyang 712046.
Department of Pathophysiology, Shaanxi University of Chinese Medicine, Xianyang 712046.
Zhongguo Ying Yong Sheng Li Xue Za Zhi. 2020 Nov;36(6):576-581. doi: 10.12047/j.cjap.6010.2020.121.
To investigate underlying mechanism involving Roumudan(RMD) formulation (Z20160012) suppressed liver fibrosis induced by CCl injection in mice by inhibiting TGF-β1/Smad4 pathway. Male BALB/c mice were randomly divided into control group, liver fibrosis model group and RMD-treated group(=11). Mice in liver fibrosis model and RMD-treaded groups were injected intraperitoneally with CCl (20% in olive oil) at the dose of 2.5 mL/kg two times for one week and 5 mL/kg two times for 4 weeks. Mice in control group were treated intraperitoneally with the same volume of olive oil at the same time intervals. From sixth week, Mice in liver fibrosis model group were administrated with CCl (20% in olive oil, 1.5 ml/kg once per week) intraperitoneally and given distilled water by intragastric gavage. Mice in the RMD-treated group were administrated with CCl (20% in olive oil, 1.5 ml/kg/mouse once per week) intraperitoneally and given RMD(6.2 g/kg everyday) by intragastric gavage. Mice in the control group were administrated with olive oil (1.5 ml/kg/mouse once per week) intraperitoneally and given distilled water by intragastric gavage. The serum AST and ALT levels were estimated for assessment of liver function. The pathologic changes of mice' livers were examined by the HE, Masson, immunohistochemical staining, Western Blot, Q-PCR and so on. After intraperitoneally injected with CCl in mice, the pathological characteristics of liver fibrosis were observed compared with the control group at the sixth week. Compared with the liver fibrosis model group, RMD improved the liver function significantly through reducing liver index(＜0.01) and the levels of ALT(＜0.01), AST(＜0.01) and HYP(＜0.05). The expression of TGF-β1(＜0.05), α-SMA(＜0.05), (＜0.01) and (＜0.01) were decreased by RMD. The positive expression area of mRNA in RMD treated group was lower than that in liver fibrosis model group. The RMD formulation could attenuate liver fibrosis by inhibiting TGF-β1/Smad4 pathway and extracellular matrix (ECM) production in mice.
为探讨柔木丹（RMD）制剂（Z20160012）通过抑制TGF-β1/Smad4信号通路抑制小鼠四氯化碳（CCl）注射诱导的肝纤维化的潜在机制。将雄性BALB/c小鼠随机分为对照组、肝纤维化模型组和RMD治疗组（每组n = 11）。肝纤维化模型组和RMD治疗组小鼠腹腔注射2.5 mL/kg剂量的CCl（20%溶于橄榄油），每周2次，共1周，之后4周每周2次，每次5 mL/kg。对照组小鼠在相同时间间隔腹腔注射相同体积的橄榄油。从第6周起，肝纤维化模型组小鼠腹腔注射CCl（20%溶于橄榄油，1.5 ml/kg，每周1次），并通过灌胃给予蒸馏水。RMD治疗组小鼠腹腔注射CCl（20%溶于橄榄油，1.5 ml/kg/只，每周1次），并通过灌胃给予RMD（6.2 g/kg/天）。对照组小鼠腹腔注射橄榄油（1.5 ml/kg/只，每周1次），并通过灌胃给予蒸馏水。检测血清AST和ALT水平以评估肝功能。通过HE染色、Masson染色、免疫组化染色、Western Blot、Q-PCR等方法检测小鼠肝脏的病理变化。在小鼠腹腔注射CCl后，于第6周观察肝纤维化的病理特征。与肝纤维化模型组相比，RMD通过降低肝指数（P＜0.01）、ALT水平（P＜0.01）、AST水平（P＜0.01）和HYP水平（P＜0.05）显著改善了肝功能。RMD降低了TGF-β1（P＜0.05）、α-SMA（P＜0.05）、Smad2（P＜0.01）和Smad3（P＜0.01）的表达。RMD治疗组中Smad4 mRNA的阳性表达面积低于肝纤维化模型组。RMD制剂可通过抑制小鼠TGF-β1/Smad4信号通路和细胞外基质（ECM）生成来减轻肝纤维化。
