黄甫静 张金娟 董莉 李雕 张春雷 廖尚高 何迅

中圖分类号 R965 文献标志码 A 文章编号 1001-0408（2021）14-1685-07

DOI 10.6039/j.issn.1001-0408.2021.14.04

摘 要 目的：考察黄水枝醇提物（TPME）对四氯化碳（CCl4）致小鼠肝纤维化的改善作用，并初步探讨其可能的作用机制。方法：将60只雄性昆明种小鼠按体质量随机分为正常组、模型组、阳性对照组（秋水仙碱0.1 mg/kg）和TPME低、中、高剂量组（250、500、1 000 mg/kg），每组10只。除正常组外，其余各组小鼠均腹腔注射20%CCl4橄榄油溶液诱导肝纤维化，每周2次，连续8周。从造模的第5周起，各给药组小鼠灌胃相应药液，正常组和模型组小鼠灌胃等体积生理盐水，每天1次，连续4周。末次给药12 h后，称定各组小鼠肝脏质量并计算肝指数，检测血清中丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）、超氧化物歧化酶（SOD）活性和丙二醛（MDA）、Ⅲ型前胶原（PC-Ⅲ）、Ⅳ型胶原（C-Ⅳ）、层粘连蛋白（LN）、肿瘤坏死因子α（TNF-α）、白细胞介素6（IL-6）含量;采用Western blot法检测肝组织中α-平滑肌肌动蛋白（α-SMA）、转化生长因子β1（TGF-β1）和Smad3蛋白的表达水平;以苏木精-伊红（HE）、Masson染色后观察肝组织病理变化。结果：与正常组比较，模型组小鼠肝指数和血清中ALT、AST活性以及MDA、PC-Ⅲ、C-Ⅳ、LN、TNF-α、IL-6含量均显著升高，SOD活性显著降低（P<0.01），肝组织中α-SMA、TGF-β1和Smad3蛋白的表达水平均显著升高（P<0.01），镜下可见肝组织出现明显的纤维化病变。与模型组比较，阳性对照组和TPME各剂量组小鼠肝指数和血清中ALT、AST活性以及MDA、PC-Ⅲ、C-Ⅳ、LN、TNF-α、IL-6含量均显著降低，SOD活性均显著升高（P<0.05或P<0.01），肝组织中α-SMA、TGF-β1和Smad3蛋白的表达水平均显著降低（P<0.05或P<0.01），镜下可见肝纤维化病变均有不同程度的改善。与TPME低剂量组比较，TPME高剂量组小鼠血清中PC-Ⅲ、LN、IL-6含量和肝组织中TGF-β1、Smad3蛋白的表达水平均显著降低（P<0.05）。结论：TPME对CCl4诱导的小鼠肝纤维化具有一定的改善作用，该作用可能与抑制胶原合成、抑制氧化应激反应、降低炎症因子含量、下调α-SMA和TGF-β1/Smad信号通路相关蛋白的表达有关。

关键词 黄水枝;肝纤维化;炎症反应;氧化应激;转化生长因子β1/Smad信号通路;小鼠

Preliminary Study on Improvement Effect of Tiarella polyphylla Ethanol Extract on CCl4-induced Hepatic Fibrosis in Mice and Its Mechanism

HUANG Fujing1，ZHANG Jinjuan2，DONG Li1，LI Diao1，ZHANG Chunlei1，LIAO Shanggao1，HE Xun1（1. School of Pharmacy， Guizhou Medical University， Guiyang 550025， China; 2. School of Basic Medicine， Guizhou Medical University， Guiyang 550025， China）

ABSTRACT   OBJECTIVE： To investigate the improvement effect of Tiarella polyphylla ethanol extract （TPME） on CCl4-induced hepatic fibrosis in mice， and to explore its possible mechanism preliminarily. METHODS： Totally 60 male Kunming mice were randomly divided into normal group， model group， positive control group （colchicine 0.1 mg/kg）， TPME low-dose， medium-dose and high-dose groups （250， 500， 1 000 mg/kg） according to body weight， with 10 mice in each group. Except for normal group， other groups were given 20% CCl4 olive oil solution intraperitoneally to induce hepatic fibrosis， twice a week， for consecutive 8 weeks. From the fifth week after modeling， administration groups were given relevant medicine intragastrically， normal group and model group were given constant volume of normal saline intragastrically， once a day， for consecutive 4 weeks. Twelve hours after last administration， the liver weight of mice in each group was measured and the liver index was calculated. The serum contents of ALT， AST， SOD， MDA， PC-Ⅲ， C-Ⅳ， LN， TNF-α and IL-6 were determined. Western blot assay was used to detect the protein expression of α-SMA， TGF-β1 and Smad3 in liver tissue. HE and Masson staining were used to observe the pathological changes of hepatic tissue. RESULTS： Compared with normal group， the liver index， the activities of ALT and AST and the contents of MDA， LN， PC-Ⅲ， C-Ⅳ， LN， TNF-α and IL-6 in serum were increased significantly， while the activity of SOD was decreased significantly in model group （P<0.01）; the protein expression of α-SMA， TGF-β1 and Smad3 in liver tissues were increased significantly （P<0.01）. Obvious fibrosis lesions was observed in liver tissue. Compared with model group， the live indexes， the activities of ALT and AST， the contents of MDA， PC-Ⅲ， C-Ⅳ， LN， TNF-α and IL-6 in serum were decreased significantly in positive control group and TPME groups， while the activities of SOD were increased significantly （P<0.05 or P<0.01）. The protein expression of α-SMA， TGF-β1 and Smad3 in liver tissue were decreased significantly （P<0.05 or P<0.01）， and liver fibrosis was improved to different extent. Compared with TPME low-dose group， the contents of PC-Ⅲ， LN and IL-6  in serum， protein expression of TGF-β1 and Smad3 in liver tissue were decreased significantly in TPME high-dose group （P<0.05）. CONCLUSIONS： TPME can improve hepatic fibrosis induced by CCl4 in mice， the mechanism of which may be associated with the inhibition of collagen synthesis and oxidative stress， the reduction of inflammatory factors， and the down-regulation of the expression α-SMA and relative proteins of TGF-β1/Smad signal pathway.

KEYWORDS   Tiarella polyphylla; Hepatic fibrosis; Inflammation response; Oxidative stress; TGF-β1/Smad signal pathway; Mice

肝纤维化（hepatic fibrosis）是肝脏遭到病毒、自由基、炎症因子等损伤因素攻击时出现的损伤-修复反应，是各种慢性肝病向肝硬化乃至肝癌发展的中间环节[1]。研究表明，肝纤维化具有可逆性[2]，抑制并逆转肝纤维化对阻止慢性肝病向肝硬化、肝癌发展具有重要意义。

目前，临床上尚无有效的肝纤维化治疗药物[3]。……