肝宁方对NASH小鼠保护作用及大鼠肝星状细胞影响的实验研究  

Protective effect of Ganning prescription on mice with nonalcoholic steatohepatitis and its effect on rat hepatic stellate cells:An experimental study

作　　者：彭佩纯 邓鑫 PENG Peichun;DENG Xin(Guangxi International Zhuang Medicine Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning 530001,Guangxi,China;School of Basic Medicine,Guangxi University of Chinese Medicine,Nanning 530004,Guangxi,China)

机构地区：[1]广西中医药大学附属国际壮医医院,广西南宁530001 [2]广西中医药大学基础医学院,广西南宁530004

出　　处：《湖南中医杂志》2023年第6期197-204,共8页Hunan Journal of Traditional Chinese Medicine

基　　金：广西壮族自治区自然科学基金项目(2021GXNSFBA220037);广西壮族自治区高校中青年教师科研基础能力提升项目(2019KY0334);广西中医药大学研究生教育创新计划项目(YCBXJ2021020);广西中医药大学桂派中医药传承创新团队项目(2022B004);广西国际壮医医院课题(2019013)。

摘　　要：目的:探讨肝宁方对非酒精性脂肪性肝炎(NASH)小鼠的保护作用及其对活化后大鼠肝星状细胞(HSCs)的影响。方法:将24只小鼠随机分为正常组、模型组、肝宁方组,每组各8只。对模型组、肝宁方组采用高脂饮食联合四氯化碳豆油溶液皮下注射建立NASH小鼠模型,造模成功后正常组和模型组予0.9%氯化钠注射液灌胃,肝宁方组予肝宁方灌胃。干预3周。将大鼠HSCs分成正常组、模型组及肝宁方组,正常组予正常培养基培养,模型组及肝宁方组采用转化生长因子-β1(TGF-β1)活化,随后正常组和模型组用含正常大鼠血清培养基培养,肝宁方组加入含30%肝宁方含药血清培养基培养。培养24 h。观察肝组织病理学情况;血清天冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT)、血清总胆固醇(CHOL)、三酰甘油(TG)水平;肝组织中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)表达,葡萄糖调节蛋白78(GRP78)及内质网核信号转导蛋白1α(IRE-1α)蛋白表达;肝细胞凋亡情况;HSCs的增殖情况。结果:肝宁方组和模型组小鼠相比,脂质空泡和出血减少,炎性细胞浸润减轻。小鼠血清AST、ALT、CHOL、TG水平,肝组织TNF-α、IL-6水平,GRP78、IRE-1α蛋白表达,肝细胞凋亡情况模型组与正常组比较,肝宁方组与模型组比较,差异均有统计学意义(P<0.01或P<0.05)。与模型组相比,肝宁方含药血清干预后HSCs增殖趋势明显减弱(P<0.01)。结论:肝宁方能够改善NASH小鼠肝脏病理学变化,保护肝脏、调节脂质代谢、抑制炎症反应、减轻肝细胞凋亡、抑制HSCs增殖,其机制可能与调控内质网应激有关。Objective:To investigate the protective effect of Ganning prescription on mice with nonalcoholic steatohepatitis(NASH) and its effect on activated rat hepatic stellate cells(HSCs).Methods:A total of 24 mice were randomly divided into normal group,model group,and Ganning prescription group,with 8 mice in each group.The mice in the model group and the Ganning prescription group were given high-fat diet combined with subcutaneous injection of carbon tetrachloride-soybean oil solution to establish a mouse model of NASH.After successful modeling,the mice in the normal group and the model group were given 0.9% sodium chloride injection by gavage,and those in the Ganning prescription group were given Ganning prescription by gavage,for 3 weeks.Rat HSCs were divided into normal group,model group,and Ganning prescription group.HSCs in the normal group were cultured in a normal medium,and those in the model group and the Ganning prescription group were activated by transforming growth factor-beta 1;then HSCs in the normal group and the model group were cultured in a medium containing normal rat serum,and those in the Ganning prescription group were cultured in a medium of serum containing 30% Ganning prescription.HSCs were cultured for 24 hours.Liver histopathology was observed;the serum levels of aspartate aminotransferase(AST),alanine aminotransferase(ALT),total cholesterol(CHOL),and triglyceride(TG) were measured,as well as the protein expression levels of tumor necrosis factor-α(TNF-α),interleukin-6(IL-6),glucose-regulated protein 78(GRP78),and IRE-1α in liver tissue;the apoptosis of hepatocytes and the proliferation of HSCs were observed.Results:Compared with the mice in the model group,the mice in the Ganning prescription group had reductions in lipid vacuoles and bleeding and alleviation of inflammatory cell infiltration.There were significant differences in the serum levels of AST,ALT,CHOL,and TG,the levels of TNF-α and IL-6 in liver tissue,the protein expression levels of GRP78 and IRE-1α,and the apoptosis of

关 键 词：非酒精性脂肪性肝炎 肝宁方 肝星状细胞 内质网应激 实验研究 

分 类 号：R285.5[医药卫生—中药学]