摘要
目的建立高脂饲料诱导的非酒精性脂肪肝(nonalcoholic fatty liver disease,NAFLD)大鼠模型,并观察其血浆摄食抑制因子nesfatin-1变化。方法雄性SPF级SD大鼠,随机分为对照组(n=6)和模型组(n=10)。对照组大鼠给予标准对照饲料,模型组则采用高脂饲料诱导NAFLD模型。葡萄糖耐量实验方法观察糖代谢变化,化学发光法检测血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、总胆红素(TBIL)、直接胆红素(DBIL)、间接胆红素(IBIL)、总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)水平,ELISA方法检测血浆nesfatin-1、胰岛素及瘦素浓度。肝脏湿重、肝指数及肝脏形态学观察,Western blot方法观察β-catenin、磷酸化β-catenin(p-β-catenin)及cyclin D1蛋白在肝脏的表达。结果与对照组比较,高脂饲料组大鼠体重、肝脏湿重和肝指数显著增加,血清ALT、TBIL、IBIL、TC、LDL-C及血浆nesfatin-1浓度显著上升,腹腔注射葡萄糖耐量实验的曲线下面积显著升高,而胰岛素及瘦素浓度则明显下降。相关性分析结果表明,血浆nesfatin-1浓度与瘦素浓度呈负相关(r=-0.571,P=0.021)。HE染色结果显示高脂饲料组大鼠肝脏呈现轻度炎症反应及程度不等的脂肪变性。Western blot结果表明高脂饲料组大鼠肝脏cyclin D1蛋白表达上调,而p-β-catenin表达则显著减少。结论高脂饲料连续喂养4周可诱导大鼠NAFLD样代谢紊乱的表现,并且nesfatin-1可能参与这一过程。
Objective To investigate the change of plasma nesfatin-1 concentration in a nonalcoholic fatty liver disease rat model induced by high-fat diet,and explore its effect on the dysfunction of glucose and lipid metabolism. Methods The nonalcoholic fatty liver disease rat model was established through introduction of a high-fat diet,and four weeks later,the intraperitoneal glucose tolerance test was conducted. Serum concentrations ofalanine aminotransferase（ ALT）, aspartate aminotransferase（ AST）, total bilirubin（ TBIL）,direct bilirubin（ DBIL）,indirect bilirubin（ IBIL）,total cholesterol（ TC）,triglyceride（ TG）, high density lipoprotein cholesterol（ HDL-C）, and low density lipoprotein cholesterol（ LDL-C） were detected using chemiluminescence technique. The plasma levels of nesfatin-1,leptin,and insulin（ INS） were measured via enzyme-linked immunosorbent assay（ ELISA）,the histological changes of the liver was observed via HE staining,and the protein expressions of β-catenin,p-β-catenin and cyclin D1 in the liver were detected using western blot and compared with β-actin. Results The bodyweight,liver weight,liver index,and area under the curve of the intraperitoneal glucose tolerance test were all higher in the model rats than those in the controls. Compared with the control,serum concentrations of ALT,TBIL,IBIL,TC and LDL-C in the model rats were all increased. The plasma nesfatin-1 level was increased in model rats while the plasma concentrations of leptin and insulin were decreased,and a negative correlation was found between the plasma concentration of nesfatin-1 and leptin. Inflammation and hepatocyte steatosis were detected in the livers of model rats,and the protein expression of cyclin D1 was upregulated while the phosphorylation of β-catenin was decreased in the livers of the model rats. Conclusion Post-creation of nonalcoholic fatty liver disease rat models through high fat diets,changes were observed in plasma nesfatin-1 concentration,perhaps a vital part of glucose and lipid metabolism dysfunction.
出处
《卫生研究》
CAS
CSCD
北大核心
2016年第3期452-457,共6页
Journal of Hygiene Research
基金
国家自然科学基金(No.81401122)
安徽省自然科学基金(No.1408085MH154)
教育部高等学校博士点基金(No.20133420120005)
