基于斑马鱼模型的疏肝降脂片改善非酒精性脂肪肝病的作用机制

Mechanism of Shugan Jiangzhi Pill improving non-alcoholic fatty liver disease based upon a zebrafishmodel

目的:研究疏肝降脂片(SGJZ)对非酒精性脂肪肝病(non-alcoholic fatty liver disease,NAFLD)斑马鱼的干预作用及其可能的机制。方法:采用2 mg·mL^(–1)蛋黄粉浸泡诱导构建NAFLD斑马鱼模型,分别给予1.63、3.26 mg·mL^(–1)SGJZ干预,连续处理3 d,油红O染色法评判脂肪肝发生率;HE染色法观察肝组织病理形态;微板法检测斑马鱼甘油三酯(triglyceride,TG)、总胆固醇(total cholesterol,TC)、谷丙转氨酶(alanine transaminase,ALT)、谷草转氨酶(aspartate transaminase,AST)、高密度脂蛋白胆固醇(high density lipoprotein cholesterol,HDL-C)、低密度脂蛋白胆固醇(low density lipoprotein cholesterol,LDL-C)含量;RT-qPCR法检测TLR4/MyD88/NF-κB信号通路相关mRNA表达水平。结果:蛋黄粉处理后,NAFLD斑马鱼模型造模成功;与Model组相比,SGJZ干预后幼鱼肝脏部位脂质沉积减轻,脂肪变性减轻;TG、TC、ALT、AST、LDL-C含量降低,HDL-C水平升高(P<0.05),呈剂量依赖性;SGJZ能下调TLR4、MyD88、IL-6、IL-1β、TNF-α、NF-κB的表达水平,高剂量下调作用更为显著(P<0.05)。结论:SGJZ对蛋黄粉诱导的NAFLD斑马鱼具有保护作用,机制可能与调控TLR4/MyD88/NF-κB信号通路有关。

OBJECTIVE To explore the interventional mechanism of Shugan Jiangzhi Pill(SGJZ)in the context of nonalcoholic fatty liver disease(NAFLD)utilizing a zebrafish model.METHODS NAFLD zebrafish model was established by immersing in egg yolk powder with SGJZ over a period of 3 consecutive days.Oil red O stain was utilized for assessing the preva⁃lence of fatty liver and hematoxylin-eosin HE stain for observing the pathological morphology of liver histology.Microplating was utilized for measuring the levels of triglyceride(TG),total cholesterol(TC),alanine aminotransferase(ALT),aspartate trans⁃aminase(AST)and low-density lipoprotein cholesterol(LDL-C)in zebrafish.Additionally,real-time quantitative polymerase chain reaction(RT-qPCR)was employed for determining the mRNA expression levels associated with the TLR4/MyD88/NF-κB signaling pathway.RESULTS After exposure to egg yolk powder,NAFLD zebrafish model was successfully established.Comparatively,SGJZ dosing significantly lowered lipid deposition and steatosis in liver of larval NAFLD zebrafish as compared with model group.The levels of TG,TC,ALT,AST and LDL-C dropped obviously while HDL-C level spiked(P<0.05).These outcomes demonstrated a dose-dependent relationship.In comparison to model group,SGJZ exhibited the capacity of sup⁃pressing the expression levels of TLR4,MyD88,IL-6,IL-1β,TNF-αand NF-κB at higher doses(P<0.05).CONCLUSISON SGJZ has demonstrated its potential of ameliorating NAFLD in zebrafish induced by egg yolk powder.The mechanism appears to be associated with a regulation of TLR4/MyD88/NF-κB signaling pathway.