卫矛醇提取物对小鼠肝纤维化的预防作用及其机制的初步探讨  

Prevention of liver fibrosis in mice by ethanol extract of Euonymus alatus and its preliminary mechanism

作　　者：黄涵柽 张孟科 周本宏[2] 祝灿[3] 黄德斌 董卫国[1] Huang Hancheng;Zhang Mengke;Zhou Benhong;Zhu Can;Huang Debin;Dong Weiguo(Department of Gastroenterology,Renmin Hospital of Wuhan University,Wuhan 430061,China;Department of Pharmacy,Renmin Hospital of Wuhan University,Wuhan 430060,China;Management Office of Scientific Research,Dong Fureng Institute of Economic and Social Development,Wuhan University,Wuhan 430071,China;Department of Pharmacy,Hubei Minzu University,Enshi 445000,China)

机构地区：[1]武汉大学人民医院消化内科,武汉430061 [2]武汉大学人民医院药学部,武汉430060 [3]武汉大学董辅礽经济社会发展研究院科研管理办公室,武汉430071 [4]湖北民族大学药理学教研室,恩施445000

出　　处：《中华消化杂志》2022年第3期188-197,共10页Chinese Journal of Digestion

基　　金：国家自然科学基金(82060745)。

摘　　要：目的研究卫矛茎皮醇提取物(EAT)和卫矛翅翘醇提取物(EAW)抗四氯化碳诱导小鼠肝纤维化的作用,并初步探讨其可能机制。方法选取60只C57BL/6小鼠,按随机数字表法随机分成健康对照组、模型组、EAW低剂量组、EAW高剂量组、EAT低剂量组、EAT高剂量组,每组10只。从造模前3 d开始,EAT低、高剂量组和EAW低、高剂量组小鼠分别予EAT和EAW各2.0、8.0 g/kg(含生药量)灌胃,健康对照组和模型组小鼠均给予等体积纯净水灌胃,1次/d,直至开始造模后第30天,共灌胃33次。EAT低、高剂量组和EAW低、高剂量组,以及模型组小鼠均予5%四氯化碳橄榄油溶液8 mL/kg腹腔注射,健康对照组小鼠予等体积0.9%氯化钠溶液腹腔注射,每周注射2次,至开始造模后第30天,共注射9次。检测各组小鼠的肝脏指数和血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、总胆红素、白细胞介素-6(IL-6)水平。采用苏木精-伊红和Masson染色法观察小鼠肝组织病理学变化并计算胶原容积分数,采用改良组织学炎症活动度(HAI)和Ishak系统评分评估小鼠肝组织的肝脏炎症反应和纤维化程度。采用免疫组织化学染色法检测α-平滑肌肌动蛋白(α-SMA)表达水平,蛋白质印迹法检测α-SMA、基质金属蛋白酶2(MMP2)和胞外信号调节激酶(ERK)1/2的蛋白质表达水平,荧光定量聚合酶链反应检测MMP2、ERK1/2的mRNA表达水平。统计学方法采用方差分析、Tukey检验、Dunn检验。结果EAW低、高剂量组和EAT高剂量组小鼠的肝脏指数均低于模型组(0.06±0.01、0.05±0.01、0.05±0.01比0.07±0.01),差异均有统计学意义(q=5.12、7.70、7.11,均P<0.01)。EAW低、高剂量组和EAT低、高剂量组小鼠血清ALT、AST水平均低于模型组[(601.76±141.38)、(283.35±42.32)、(734.74±116.06)、(391.60±34.33)U/L比(982.45±96.04)U/L,(509.49±152.29)、(345.41±67.39)、(282.30±65.72)、(243.23±45.20)U/L比(766.01±114.49)U/L],差异均有统计学意�Objective To study the role of ethanol extract of Euonymus alatus stems(EAT)and ethanol extract of Euonymus alatus wings(EAW)in anti-hepatic fibrosis induced by carbon tetrachloride in mice,and to explore its preliminary mechanism.Methods Sixty C57BL/6 mice were selected and randomly divided into healthy control group,carbon tetrachloride model(CTM)group,EAW low dose(EAW-L)group,EAW high dose(EAW-H)group,EAT low dose(EAT-L)group and EAT high dose(EAT-H)group,with 10 mice in each group.Three days before modeling,the mice of EAT-L,EAT-H,EAW-L and EAW-H group were gavaged with EAT or EAW at 2.0 or 8.0 g/kg,respectively,and the mice of healthy control group and CTM group were gavaged with equal volume of pure water,once a day till the 30th day after modeling(total 33 times).Five percent carbon tetrachloride olive oil solution was intraperitoneally injected at 8 mL/kg to establish liver fibrosis model in CTM,EAT-L,EAT-H,EAW-L and EAW-H groups.The mice in the healthy control group were intraperitoneally injected with equal volume of 0.9%sodium chloride solution,twice per week for 30 days,and a total of 9 times of injection.The liver index,serum alanine aminotransferase(ALT),aspartate aminotransferase(AST),total bilirubin(TBil)and interleukin-6(IL-6)were detected.Hematoxylin-eosin and Masson staining were used to observe the pathological changes of mouse liver tissue and calculate the collagen volume fraction.The liver inflammatory response and fibrosis degree were evaluated by histological activity index(HAI)and Ishak system score.The level ofα-smooth muscle actin(α-SMA)in liver tissue was both detected by immunohistochemistry and Western blotting.The expression of matrix metalloproteinase 2(MMP2)and extracellular signal-regulated kinase(ERK)1/2 at protein and mRNA level was detected by Western blotting and fluorescent quantitative polymerase chain reaction.Analysis of variance,Tukey test and Dunn test were used for statistical analysis.Results The hepatic indexes of EAW-L,EAW-H and EAT-H groups were lower than that

关 键 词：卫矛醇提物 Ras/ERK-MMP2 炎症损伤 肝纤维化 

分 类 号：R285.5[医药卫生—中药学]