摘要
[目的]研究非酒精性脂肪性肝炎(non-alcoholic steatohepatitis,NASH)小鼠CXC趋化因子配体-12(CXC chemokine ligand-12,CXCL-12)/CXC趋化因子受体-4(CXC chemokine receptor-4,CXCR-4)/磷酸肌醇3-激酶(phosphoinositide 3-kinase,PI3K)/苏氨酸蛋白激酶(acid threonine protein kinase,AKT)/核因子-κB(nuclear factor-κB,NF-κB)信号通路的激活情况以及黄连素对该信号通路的影响,阐明NASH炎症反应的可能机制及黄连素的作用。[方法] 8周龄雄性ApoE基因敲除小鼠随机分成模型组、黄连素组和对照组。采用高脂高胆固醇饲料连续喂养12周建立NASH模型,第6周开始黄连素组予黄连素200mg(/kg·d)灌胃6周,第12周处死各组小鼠,检测体质量、ALT、AST、TC、TG水平;HE染色评价肝组织病理学改变;免疫组化检测小鼠肝组织CXCL-12、CXCR-4、pAKT、pNF-κB表达水平;Real-time PCR和Western blot分别检测小鼠肝脏组织CXCL-12、CXCR-4、PI3K、AKT、NF-κB的mRNA和蛋白表达水平。[结果]黄连素组ALT、AST、TC明显低于模型组(P<0.05);HE染色显示,黄连素组小鼠肝细胞气球样变、小叶内炎症较模型组明显减轻(P<0.05),NAS总分也低于模型组(P<0.05);免疫组化显示黄连素组小鼠的CXCL-12、CXCR-4、p AKT、p NF-κB阳性表达弱于模型组;CXCL-12、CXCR-4、PI3K、AKT、NF-κB mRNA以及CXCL-12、CXCR-4、p AKT、pNF-κB蛋白表达水平均低于模型组(P<0.05)。[结论]CXCL-12/CXCR-4/PI3K/AKT/NF-κB信号通路在小鼠NASH发生中发挥作用;黄连素能明显抑制NASH小鼠肝组织内CXCL-12/CXCR-4/PI3K/AKT/NF-κB信号通路的激活并能降低转氨酶和血脂,减轻肝脏内脂质沉积和炎症反应。
[Objective]To observe the activation of CXCL-12/CXCR-4/PI3K/AKT/NF-κB signaling pathway in non-alcoholic steatohepatitis(NASH)mice, and clarify the moleeular mechanism of inflammation initiated in NASH and the possible mechanism of berberine.[Methods]ApoE knockout mice(8weeks old),were assigned into the 3groups,model group,berberine group and control group randomly.NASH model was established by continuous feeding of high-fat and high-eholesterol feed for 12weeks.200mg/(kg'd)of berberberine was given to berberberine group at the 6th week for 6weeks and each group of mice was executed at the 12th week.Body weight,indicators of liver enzymes and blood lipid were detected.HE staining was used to evaluate the pathological changes.Immunohistoehemieal was used to detect CXCL-12,CXCR-4,pAKT,pNF-κB protein.Real-time PCR and Western blot was used to detect CXCL-12/CXCR-4/PI3K/AKT/NF-κB mRNA and protein expression level in mice liver tissue.[Results]ALT,AST and total cholesterol in berberine group were significantly lower than model group(P<0.05).Compared with model group,the balloon-like changes,lobular inflammation and NAS total scores were significantly decreased in berberberine group(P<0.05).The expression of CXCL-12,CXCR-4,PI3K,AKT and NF-κB mRNA in berberine group mice liver tissue were lower than model group(P<0.05).Irnmunohistochemieal result also shows the positive expression of CXCL-12, CXCR-4,pAKT,pNF-κB decreased.Western blot showed that CXCL-12,CXCR-4,pAKT and pNF-κB protein were significandy lower than model group(P<0.05).[Conelusion]CXCL-12/CXCR-4/PI3K/AKT/NF-κB signaling pathway plays a vital role in progression of NASH.Berberine can inhibit the activation of CXCL-12/CXCR-4/PI3K/AKT/NF-κB signaling pathway in the liver tissues of NASH mice and reduce liver lipid deposition,inflammation, which can be used to prevent and treat NASH.
作者
李玲
吕惠卿
LI Ling;LYU Huiqing(Zhejiang Chinese Medical University School of Nursing,Hangzhou(310053),China;College of Pharmaceutical Science,Zhcjiang Chinese Medical University)
出处
《浙江中医药大学学报》
CAS
2018年第11期941-950,共10页
Journal of Zhejiang Chinese Medical University
基金
浙江省中医药科学研究基金项目(792214E01004)
浙江省自然科学基金项目(LY16H100003)
浙江省科技厅公益技术应用(2017C33235)~~
