新橙皮苷对非酒精性脂肪性肝炎小鼠NLRP3/NF-κB信号通路的影响

Effect of Neohesperidin on NLRP3/NF-κB Signaling Pathway on mice with non-alcoholic steatohepatitis

目的探讨新橙皮苷(Neohesperidin,NHP)对非酒精性脂肪性肝炎(NASH)小鼠的治疗作用及其机制。方法24只SPF级C57BL6雄性小鼠随机分为正常组、模型组和NHP组(40mg/kg)。通过喂饲高脂高胆固醇饲料(HFHC)制备NASH小鼠模型,造模同时给予相应药物治疗,12周后进行葡萄糖耐量试验、胰岛素耐量试验,计算小鼠的体重、肝重、肝指数,采集血清和肝组织标本,检测血清中胆固醇(TC),甘油三酯(TG),丙氨酸氨基转移酶(ALT),天门冬氨酸氨基转移酶(AST);蛋白免疫印迹法(Western blot)检测肝脏中炎症小体3(NLRP3)的蛋白表达及磷酸化核转录因子-κB(NF-κB)的蛋白表达情况;苏木素-伊红(HE)染色和油红O染色观察肝脏病理形态学变化。结果与正常组比较,模型组小鼠血清中TC、TG、ALT、AST含量、肝脏中NLRP3和NF-κB的mRNA和蛋白表达及p-NF-κB的蛋白表达水平显著升高(P<0.05);与模型组比较,NHP组小鼠血清中TC、TG、ALT、AST含量、肝脏中NLRP3和NF-κB的mRNA和蛋白表达及p-NF-κB的蛋白表达水平均明显降低(P<0.05)。HE染色和油红O染色结果提示其能明显减轻肝脏脂肪变性程度。结论NHP可能通过抑制NLRP3/NF-κB信号通路,减轻炎症反应来治疗NASH。

Objective To explore the therapeutic effect and mechanism of Neohesperidin(NHP) on non-alcoholic steatohepatitis(NASH) mice.Methods 24 SPF C57BL6 male mice were randomly divided into normal group, model group and NHP group(40mg/kg). High-fat and high-cholesterol diet(HFHC) were fed to bulid the NAFLD model, and each treatment group was given corresponding drugs at the same time. After 12 weeks, conducted glucose tolerance test and insulin tolerance test, calculated mice body weight, liver weight and liver index, collected serum and liver tissue samples to detecte in total cholesterol(TC),triglyceride(TG),alanine aminotransferase(ALT), aspartic acid aminotransferase(AST). Real-time fluorescence quantitative polymerase chain reaction(Real-time PCR) and Western blot were used to observe the gene and protein expressions of inflammatory body 3(NLRP3) and protein expression of phosphorylated nuclear transcription factor-κB(NF-κB) in liver.Hematoxylin-eosin(HE) staining and Oil red staining to observe the pathological morphological changes of liver. Results Compared with normal group, the contents of TC, TG, ALT and AST in serum, the mRNA and protein expression of NLRP3 and NF-κB in liver and the protein expression of P-NK-κB in model group were significantly increased(P<0.05).Compared with model group, the contents of TC, TG, ALT and AST in serum, the mRNA and protein expression of NLRP3 and NF-κB in liver and the protein expression of P-NF-κB in NHP group were distinctly decreased(P<0.05).HE staining and Oil red staining indicated that NHP could significantly reduce the degree of liver steatosis.Conclusion NHP may treat NASH by inhibiting NLRP3/NF-κB signaling pathway and alleviating inflammatory response.