摘要
目的 探讨复方茵芪三黄解毒汤抗肝纤维化进展的可能作用机制。方法 将38只C57BL/6J小鼠随机分为空白组、模型组、水飞蓟宾组和茵芪三黄解毒汤低、中、高剂量组,模型组8只,其余每组6只。除空白组外其余各组小鼠均腹腔注射10%四氯化碳(CCl4)建立肝纤维化模型,每周2次,持续8周。首次造模1周后开始药物干预,茵芪三黄解毒汤低、中、高剂量组分别给予茵芪三黄解毒汤混悬液8.325、16.65、33.3 g/(kg·d)灌胃,水飞蓟宾组给予水飞蓟宾混悬液55 mg/(kg·d)灌胃,空白组和模型组给予0.2 ml生理盐水灌胃,持续8周。使用小动物超声探测仪观察活体小鼠肝脏硬度记录超声灰度值,采用天狼猩红染色观察肝脏组织并计算胶原纤维沉积比例,检测各组小鼠肝功能[包括丙氨酸氨基转移酶(ALT)、门冬氨酸氨基转移酶(AST)]及血清中层粘连蛋白(LN)、透明质酸(HA)、Ⅲ型前胶原(PCⅢ)、Ⅳ型胶原(CⅣ)含量,检测小鼠肝组织中转化生长因子β1(TGF-β1)、波形蛋白(Vimentin)、E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、α平滑肌肌动蛋白(α-SMA)蛋白表达。结果 与空白组相比,模型组小鼠肝脏超声灰度值、胶原纤维沉积比例升高,血清ALT、AST、HA、LN、PCⅢ水平升高,肝组织中E-cadherin表达减少,N-cadherin、α-SMA、Vimentin、TGF-β1表达增加(P<0.05)。与模型组比较,水飞蓟宾组与茵芪三黄解毒汤中、高剂量组肝脏超声灰度值及胶原纤维沉积比例下降,血清ALT、AST、LN、HA、PCⅢ水平降低;而与模型组相比,水飞蓟宾组和茵芪三黄解毒汤各剂量组小鼠肝组织中E-cadherin表达升高,N-cadherin、Vimentin、TGF-β1、α-SMA表达均下降(P<0.05),其中茵芪三黄解毒汤中剂量组各项指标改善最为显著(P<0.05)。结论 茵芪三黄解毒汤能够显著减轻CCl4致小鼠肝纤维化的程度,可能与其调控肝脏硬度抑制小鼠上皮间质转化发生有关。
Objective To investigate the possible mechanism of compound Yinqi Sanhuang Jiedu Decoction(茵芪三黄解毒汤,YSJD)against the progression of hepatic fibrosis(HF).Methods Thirty-eight C57BL/6J mice were randomly divided into blank group,model group,silybin group and low-,medium-and high-dose YSJD groups,with eight mice in the model group and six mice each in other groups.Except for the blank group,all mice were injected in⁃traperitoneally with 10%carbon tetrachloride(CCl4)to establish a HF model,twice a week for 8 weeks.The drug in⁃tervention started one week after the first modeling;the low-,medium-and high-dose YSJD groups were given 8.325,16.65 and 33.3 g/(kg·d)of YSJD suspension by gavage,respectively,while the silybin group was given 55 mg/(kg·d)of silybin suspension by gavage,and the blank group and the model group were given 0.2 ml normal saline by gavage,all for 8 weeks.The liver hardness of living mice was observed using a small animal ultrasound detector,and greyscale ultrasound was recorded.The liver tissue was observed by Sirius scarlet staining,and the proportion of collagen fiber deposition was calculated.Liver function indicators including alanine aminotransferase(ALT)and aspartate aminotransferase(AST),and serum laminin(LN),hyaluronic acid(HA),pre-collagen type III(PCIII)and colla⁃gen type IV(CIV)were also detected.The protein expression of transforming growth factorβ1(TGF-β1),Vimentin,E-cadherin,αsmooth muscle actin(α-SMA)in liver tissue were detected.Results Compared to the blank group,the model group showed increased gray value,collagen deposition,serum ALT,AST,HA,LN and PCIII levels,de⁃creased expression of E-cadherin,and increased expression of N-cadherin,α-SMA,Vimentin and TGF-β1 in liver tissues(P<0.05).Compared to the model group,the ultrasonic gray value and the proportion of collagen fiber depo⁃sition in liver of silybin group and YSJD medium-and high-dose groups decreased,and the serum ALT,AST,LN,HA and PCⅢlevels decreased.Compared to the model group,the expressi
作者
王宁
吕文良
武庆娟
WANG Ning;LYU Wenliang;WU Qingjuan(Guang'anmen Hospital,China Academy of Chinese Medical Sciences,Beijing,100053)
出处
《中医杂志》
CSCD
北大核心
2023年第17期1792-1798,共7页
Journal of Traditional Chinese Medicine
基金
国家重点研发计划(2018YFC1705700)
国家自然科学基金(81774282)
中国中医科学院科技创新工程(C12021A00801,C12021A00802)
北京市自然基金(7222295)。
关键词
肝纤维化
茵芪三黄解毒汤
肝脏硬度
上皮间质转化
hepatic fibrosis
Yinqi Sanhuang Jiedu Decoction(茵芪三黄解毒汤)
liver hardness
epithelial mesen⁃chymal transition
