苦瓜提取物调节ZDF糖尿病大鼠糖异生的作用机制  

Mechanism of Momordica charantia Extract in Regulating Gluconeogenesis in ZDF Diabetes Rats

作　　者：许光远 张茁[2] 张晓明[1] XU Guangyuan;ZHANG Zhuo;ZHANG Xiaoming(Fuxing Hospital,Capital Medical University,Beijing 100045,China;The First Affiliated Hospital of Shaanxi University of Chinese Medicine,Xianyang 712046,China)

机构地区：[1]首都医科大学附属复兴医院,北京100045 [2]陕西中医药大学第一附属医院,陕西咸阳712046

出　　处：《中国实验方剂学杂志》2023年第16期65-71,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金青年基金项目(82104614);北京市自然科学基金项目(7204268);北京市中医药科技发展基金项目(QN2018-35018);北京中医药薪火传承3+3工程张晓明基层老中医传承工作室;北京市临床中医重点专科培育项目。

摘　　要：目的:观察苦瓜提取物对糖尿病大鼠糖异生信号通路的影响。方法:5~6周龄雄性Zucker Diabetic Fatty(ZDF)大鼠随机分为模型组、苦瓜组(苦瓜提取物0.40 g·kg^(-1)灌胃),另7只健康雄性ZDF(fa/+)大鼠为正常组,每日1次灌胃,连续6周。实验过程中,观察大鼠一般情况,记录体质量,第1、3、5周检测空腹血糖、随机血糖;第6周行口服葡萄糖耐量实验(OGTT)、检测大鼠血清甘油三酯(TG)、游离脂肪酸(FFA)、总胆固醇(TC)、天门冬氨酸氨基转移酶(AST)、丙氨酸氨基转移酶(ALT);肝苏木素-伊红(HE)染色检测肝形态结构;肝糖原染色(PAS)检测肝糖原存储;采用实时荧光定量聚合酶链式反应(Real-time PCR)检测肝脏磷酸稀醇式丙酮酸羧激酶(PEPCK)、葡萄糖-6-磷酸酶(G6Pase)mRNA表达;蛋白免疫印迹法(Western blot)检测肝脏叉头转录因子1(FoxO1)磷酸化水平及磷酸稀醇式丙酮酸羧激酶(PEPCK)、G6Pase蛋白表达。结果:与模型组比较,苦瓜组大鼠体质量、空腹血糖、随机血糖、糖耐量明显改善(P<0.05,P<0.01);血清FFA、TC、TG明显降低(P<0.05,P<0.01);ALT、AST各组间差异无统计学意义;HE显示肝细胞排列较整齐,脂肪样变性减轻;PAS显示肝糖原储存增加;肝p-FoxO1蛋白表达显著升高(P<0.01),FoxO1蛋白表达无显著差异,PEPCK、G6Pase mRNA及蛋白表达明显降低(P<0.05)。结论:苦瓜提取物具有降糖、降脂、改善糖耐量及肝糖原储备作用,与其通过上调FoxO1磷酸化来抑制PEPCK、G6Pase表达,调控糖异生相关。Objective:To observe the effect of Momordica charantia extract(MCE)on the gluconeogenesis signaling pathway in diabetes rats.Method:Male Zucker Diabetic Fatty(ZDF)rats aged 5-6 weeks were randomly divided into a model group and an MCE group(administered MCE at a dose of 0.40 g·kg^(-1) by gavage).Additionally,seven healthy male ZDF(fa/+)rats were assigned to the normal group and received administration once daily for six consecutive weeks.During the experiment,the general condition of the rats was observed,and body weight was recorded.Fasting blood glucose and random blood glucose levels were measured in the 1st,3rd,and 5th weeks.In the 6th week,an oral glucose tolerance test(OGTT)was conducted,and serum levels of triglycerides(TG),free fatty acid(FFA),total cholesterol(TC),aspartate aminotransferase(AST),and alanine aminotransferase(ALT)were measured.Hematoxylin-eosin(HE)staining was performed to examine liver morphology,periodic acid-Schiff(PAS)staining was used to assess hepatic glycogen storage,and Real-time polymerase chain reaction(PCR)was employed to measure the mRNA expression of phosphoenolpyruvate carboxykinase(PEPCK)and glucose-6-phosphatase(G6Pase)in the liver.Western blot analysis was conducted to measure the phosphorylation level of forkhead box protein O1(FoxO1)and the protein expression of PEPCK and G6Pase in the liver.Result:Compared with the model group,the MCE group showed significant improvements in body weight,fasting blood glucose,random blood glucose,and glucose tolerance(P<0.05,P<0.01)and reduced serum levels of FFA,TC,and TG(P<0.05,P<0.01).There were no significant differences in ALT and AST between the two groups.In the MCE group,the HE staining revealed more orderly liver cell arrangement and reduced hepatic steatosis and the PAS staining showed increased hepatic glycogen storage.The protein expression of p-FoxO1 in the liver was significantly elevated(P<0.01),while there was no significant difference in FoxO1 protein expression.The mRNA and protein expression of PEPCK and G6Pase signif

关 键 词：苦瓜提取物 糖尿病 糖异生 叉头转录因子1 磷酸稀醇式丙酮酸羧激酶 葡萄糖-6-磷酸酶 

分 类 号：R2-0[医药卫生—中医学] R22R242R2-031R285.5