摘要
目的探讨桃红四物汤对CCl_(4)诱导肝纤维化小鼠模型的干预作用及其作用机制。方法24只雄性C57/BL6小鼠随机分为对照组、模型组和桃红四物汤组,每组8只。模型组和桃红四物汤组小鼠腹腔注射10%CCl_(4),第3周开始给予桃红四物汤灌胃给药4周,检测肝功能(ALT、AST),观测肝组织病理形态学。实时荧光定量PCR检测α-平滑肌肌动蛋白(α-SMA)、透明质酸合成酶-2(HAS-2)、Ⅰ型胶原(Col1)mRNA的表达,蛋白免疫印迹法检测α-SMA、Col1及HAS-2蛋白的表达。分离小鼠原代肝星状细胞(HSC),应用siRNA沉默HAS-2,检测其对HSC激活的影响。计量资料两组间比较采用t检验,多组间比较采用单因素方差分析,组内进一步两两比较采用SNK或LSD-t检验。结果与正常组相比,模型组血清肝功能指标ALT、AST水平显著升高,桃红四物汤组小鼠血清ALT、AST水平均显著下降(P值均<0.01)。病理学染色显示:模型组炎症细胞浸润明显,增生的胶原纤维形成纤维间隔;桃红四物汤组胶原纤维间隔较疏松及炎症细胞浸润减轻。与模型组相比,桃红四物汤组α-SMA、Col1 mRNA和蛋白的表达水平均显著降低(P值均<0.001)。与正常组比较,模型组肝组织中HAS-2 mRNA表达水平显著升高(t=6.14,P<0.05),桃红四物汤组HAS-2蛋白表达水平(0.29±0.10)较模型组(1.00±0.12)明显减少(t=70.73,P<0.001)。siRNA沉默HAS-2,TGFβ处理后(Si HAS-2+TGFβ组)α-SMA、Col1 mRNA表达水平较NC+TGFβ组显著降低(P<0.01)。结论桃红四物汤通过抑制HAS-2对CCl 4致小鼠纤维化有显著治疗作用。
Objective To investigate the therapeutic effect of Taohong Siwu decoction on a mouse model of carbon tetrachloride(CCl_(4))-induced liver fibrosis and its mechanism of action.Methods A total of 24 male C57BL/6 mice were randomly divided into normal group,model group,and Taohong Siwu decoction group,with 8 mice in each group.The mice in the model group and the Taohong Siwu decoction group were given intraperitoneal injection of 10% CCl_(4),and Taohong Siwu decoction was given by gavage since week 3 for 4 consecutive weeks.Liver function[alanine aminotransferase(ALT)and aspartate aminotransferase(AST)]was measured,and liver pathomorphology was observed.Real-time PCR was used to measure the mRNA expression ofα-smooth muscle actin(α-SMA),hyaluronic acid synthase-2(HAS-2),and collagen typeⅠ(Col1),and Western blotting was used to measure the protein expression ofα-SMA,Col1,and HAS-2.Primary hepatic stellate cells(HSCs)were isolated,and HAS-2 was silenced by siRNA to observe its influence on HSC activation.The t-test was used for comparison of continuous data between two groups;a one-way analysis of variance was used for comparison between multiple groups,and the SNK or least significant difference t-test was used for further comparison between two groups.Results Compared with the normal group,the model group had significant increases in serum liver function parameters(ALT,AST)and the Taohong Siwu decoction group had significant reductions in the serum levels of ALT and AST(all P<0.01).Pathological staining showed that the model group had marked inflammatory cell infiltration and formation of fibrous septa by proliferated collagen fibers,and the Taohong Siwu decoction group had loose fibrous septa and alleviated inflammatory cell infiltration.Compared with the model group,the Taohong Siwu decoction group had significant reductions in the mRNA and protein expression ofα-SMA and Col1(all P<0.001).Compared with the normal group,the model group had a significant increase in the mRNA expression level of HAS-2 in liver
作者
刘旭凌
杨广越
张玮
孙田甜
马文婷
吴柳
薛冬英
刘成
陶乐
LIU Xuling;YANG Guangyue;ZHANG Wei;SUN Tiantian;MA Wenting;WU Liu;XUE Dongying;LIU Cheng;TAO Le(Laboratory of Liver Diseases,Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200062,China;Department of Infectious Diseases,Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200062,China;Central Laboratory,Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200062,China)
出处
《临床肝胆病杂志》
CAS
北大核心
2021年第11期2563-2568,共6页
Journal of Clinical Hepatology
基金
国家自然科学基金项目(81673788,81873136,81803898,81803232,82004106)
上海市自然基金(20ZR1450300)
上海市卫生健康委员会卫生行业临床研究专项(202040149)
上海市普陀区自主创新项目(ptkwws201817,ptkwws201904)。
