拳卷地錢總黃酮抗小鼠肝纖維的作用及機制研究

梁爽 朱華 黃健軍 張文濤 盧森華 陳詩曼

中圖分類號 R285.5 文獻標志碼 A 文章編號 1001-0408（2021）19-2363-08

DOI 10.6039/j.issn.1001-0408.2021.19.10

摘 要 目的：研究拳卷地錢總黃酮抗小鼠肝纖維化的作用及可能機制。方法：將72只小鼠隨機分為空白組、模型組、陽性對照組（秋水仙堿0.2 mg/kg）和卷拳地錢總黃酮高、中、低劑量組（300、150、75 mg/kg），每組12只。除空白組外，其余各組小鼠均于背部皮下注射25%四氯化碳-花生油溶液復制肝纖維化模型;于造模的同時，空白組和模型組小鼠灌胃水，其余各組小鼠灌胃相應藥物，灌胃體積均為20 mL/kg，每天1次，連續10周。末次灌胃后，檢測小鼠血清中丙氨酸轉氨酶（ALT）和天冬氨酸轉氨酶（AST）的水平;觀察小鼠肝組織病理學變化;檢測小鼠肝組織中Ⅰ型膠原蛋白（COL-Ⅰ）、COL-Ⅲ、轉化生長因子β1（TGF-β1）水平;檢測小鼠肝組織中α-平滑肌肌動蛋白（α-SMA）、TGF-β1、Smad2、Smad4、Smad7蛋白的表達水平;檢測小鼠肝組織中TGF-β1、Smad2、Smad4、Smad7 mRNA的表達水平。結果：與空白組比較，模型組小鼠血清中ALT、AST水平，肝組織中COL-Ⅰ、COL- Ⅲ、TGF-β1水平以及α-SMA 、TGF-β1、Smad2、Smad4 的蛋白表達水平和TGF-β1、Smad2、Smad4的 mRNA表達水平均顯著升高（P<0.05或P<0.01），肝組織中Smad7的mRNA及其蛋白的表達水平均顯著降低（P<0.05）;肝組織損傷及膠原纖維增生較明顯。與模型組比較，陽性對照組和拳卷地錢總黃酮高劑量組小鼠上述指標均顯著逆轉（P<0.05或P<0.01）;拳卷地錢總黃酮中劑量組小鼠血清中ALT水平，肝組織中COL-Ⅰ水平和TGF-β1、Smad2、Smad4 的mRNA及其蛋白的表達水平均顯著降低（P<0.05或P<0.01）;拳卷地錢總黃酮低劑量組小鼠肝組織中Smad2、Smad4蛋白的表達水平均顯著降低（P<0.05）;各給藥組小鼠肝組織的損傷及纖維化程度均有所減輕。結論：拳卷地錢總黃酮具有抗小鼠肝纖維化的作用，其作用機制可能與調節TGF-β/Smad信號通路中TGF-β1、Smad2、Smad4、Smad7的mRNA及其蛋白的表達相關。

關鍵詞 拳卷地錢;總黃酮;肝纖維化;TGF-β/Smad信號通路;小鼠

Study on the Effect and Mechanism of Total Flavonoids from Marchantia convoluta on the Anti-hepatic Fibrosis in Mice

LIANG Shuang1，2，ZHU Hua1，2，HUANG Jianjun1，2，ZHANG Wentao2，LU Senhua3，CHEN Shiman2（1. School of Ethnology and Sociology， Guangxi University for Nationalities， Nanning 530006， China; 2. College of Pharmacy， Guangxi University of TCM， Nanning 530200， China; 3. Food Laboratory， Yulin Center for Food and Drug Control， Guangxi Yulin 537000， China）

ABSTRACT ? OBJECTIVE： To study the effect and potential mechanism of the total flavonoids from Marchantia convoluta on anti-hepatic fibrosis in the mice. METHODS： Seventy-two mice were randomly divided into blank group， model group， positive control group （colchicine 0.2 mg/kg） and M. convoluta total flavonoids high-dose， medium-dose and low-dose groups （300， 150， 75 mg/kg）， with 12 mice in eac group. Except for blank group， other groups were subcutaneously given 25% CCl4-peanut oil solution on the back to induce liver fibrosis model. At the same time， blank group and model group were given water intragastrically， while other groups were given relevant medicine intragastrically 20 mL/kg， once a day， for consecutive 10 weeks. After last administration， the serum levels of ALT and AST were detected. Histopathological changes of liver tissue in mice was observed. The levels of ?COL-Ⅰ， COL-Ⅲ and TGF-β1 in liver tissue were detected. The protein expression levels of α-SMA and TGF-β1， Smad2， Smad4 and Smad7 in liver tissue were detected. The expression levels of TGF-β1， Smad2， Smad4 and Smad7 mRNA in liver tissue were detected. RESULTS： Compared with blank group， the serum levels of ALT and AST in model group， the levels of COL-Ⅰ， COL-Ⅲ and TGF-β1 in liver tissue， protein expression levels of α-SMA， TGF-β1， Smad2 and Smad4， mRNA expression levels of TGF-β1， Smad2 and Smad4 were increased significantly （P<0.05 or P<0.01）. The mRNA and protein expression levels of Smad7 in liver tissue were decreased significantly （P<0.05）. The degree of liver tissue injury and collagen fiber hyperplasia were serious. Compared with model group， above indexes of mice were reversed significantly in positive control group and M. convoluta total flavonoids high-dose group （P<0.05 or P<0.01）. Serum level of ALT， the levels of COL-Ⅰ， mRNA and protein expression of TGF-β1， Smad2 and Smad4 in liver tissue were decreased significantly in M. convoluta total flavonoids medium-dose group （P<0.05 or P<0.01）. Protein expression of Smad2 and Smad4 in liver tissue were decreased significantly in M. convoluta total flavonoids low-dose group （P<0.05）. The liver injury and fibrosis of mice were relieved in administration groups. CONCLUSIONS： M. convoluta total flavonoids possess the effect of anti-hepatic fibrosis， the mechanism of which is related to the regulation of mRNA and protein expression of TGF-β1， Smad2， Smad4 and Smad7 in the signaling pathway of TGF-β/Smad.