基于PPAR-α/CPT-1信号通路的桑叶总黄酮调控T2DM大鼠肝脏脂质代谢作用及其机制  

Total Flavonoids of Mulberry Leaves Improves Liver Lipid Metabolism in Type 2 Diabetic Rats by Regulating PPAR-α/CPT-1 Pathway

作　　者：段钰卉 代泓钰 安永铖 程龙 石璐 李慧敏 王晨 吕迎兰 何昶昊 张慧琳 黄艳 付宛鑫 孟闫燕 赵保胜[1] DUAN Yuhui;DAI Hongyu;AN Yongcheng;CHENG Long;SHI Lu;LI Huimin;WANG Chen;LYU Yinglan;HE Changhao;ZHANG Huilin;HUANG Yan;FU Wanxin;MENG Yanyan;ZHAO Baosheng(College of Chinese Materia Medica and Life Sciences,College of Life Sciences,Beijing Research Institute of Chinese Medicine,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]北京中医药大学中药学院生命科学学院中医药研究院,北京100029

出　　处：《中国实验方剂学杂志》2022年第15期61-69,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81973535、81773960)。

摘　　要：目的:观察桑叶总黄酮改善2型糖尿病(T2DM)大鼠肝脏脂代谢紊乱的药效学作用,并基于肝脏过氧化物酶增殖物激活受体-α(PPAR-α)、肉毒碱棕榈酰基转移酶-1(CPT-l)蛋白探讨其作用机制。方法:采用醇提法+大孔树脂纯化法提取、纯化桑叶总黄酮,并进行鉴定。利用高脂肪饮食(HFD)+链脲佐菌素(STZ)法建立T2DM大鼠模型,选择血糖≥11.1 mmol·L^(-1)的大鼠,以桑叶总黄酮高、中、低剂量(300、150、75 mg·kg^(-1))分别灌胃给药8周,观察大鼠体质量及血糖情况。苏木素-伊红(HE)染色观察大鼠肝脏病理变化,生化法检测大鼠血清脂代谢指标总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)及高密度脂蛋白胆固醇(HDL-C)水平,采用实时荧光定量聚合酶链式反应(Real-time PCR)及蛋白免疫印迹法(Western blot)检测肝脏组织PPAR-α及CPT-1 mRNA和蛋白的表达。结果:桑叶总黄酮干预8周后,与正常组比较,模型组大鼠摄食量、肝脏指数、空腹血糖显著升高(P<0.01);与模型组比较,桑叶总黄酮高、中、低剂量组大鼠摄食量、空腹血糖、肝脏指数显著降低(P<0.01)。HE结果显示,正常组大鼠肝组织结构完整未见明显异常;模型组大鼠肝细胞空泡变性且有炎性浸润;桑叶总黄酮高、中、低剂量组大鼠肝脏结构未见明显异常。血脂四项结果显示,与正常组比较,模型组大鼠TC、TG、LDL-C水平显著升高(P<0.01),HDL-C水平显著降低(P<0.01);与模型组比较,各给药组TC、TG、LDL-C水平明显降低(P<0.05,P<0.01),HDL-C水平显著增加(P<0.01)。Real-time PCR结果显示,与正常组比较,模型组大鼠PPAR-α和CPT-1 mRNA表达显著降低(P<0.01);与模型组比较,桑叶总黄酮高剂量组PPAR-α和CPT-1 mRNA表达显著升高(P<0.01)。Western blot结果显示,与正常组比较,模型组大鼠PPAR-α和CPT-1蛋白表达显著降低(P<0.01);与模型组比较,桑叶总黄酮高剂量组PPAR-α和CPT-1蛋白表达明显升高(P<0.Objective:To investigate the medicinal effect of total flavonoids of mulberry leaves on regulating liver lipid metabolism disorder in diabetes mellitus type 2(T2DM)rats,and the mechanism based on liver peroxidase proliferators activate receptors-α(PPAR-α) and carnitine palmityl transferase-1(CPT-1)proteins.Method:Total flavonoids of mulberry leaves were extracted and purified by ethanol extraction +macroporous resin purification and then identified.T2DM rat model was induced by high fat diet(HFD)+streptozocin(STZ)method.Rats with blood glucose ≥ 11.1 mmol·L^(-1)were divided into three administration groups with the high dose(300 mg·kg^(-1)),medium dose(150 mg·kg^(-1)),and low dose(75 mg·kg^(-1))of total flavonoids of mulberry leaves for 8 weeks,respectively,to observe the weight and blood glucose of the rats.The pathological changes of rat livers were observed by hematoxylin-eosin(HE)staining.Biochemical method was used to detect the levels of total cholesterol(TC),triglyceride(TG),low density lipoprotein-cholesterol(LDL-C),and high density lipoprotein-cholesterol(HDL-C)of blood lipid metabolism in rats.The messenger ribonucleic acid(mRNA) and protein expressions of PPAR-α and CPT-1 were determined by real-time quantitative polymerase chain reaction(Real-time PCR) and Western blot.Result:After 8 weeks of intervention of total flavonoids of mulberry leaves,compared with the control group,the food intake,liver index,and fasting blood glucose of rats in the model group increased significantly(P<0.01).Compared with the model group,the food intake,fasting blood glucose,and liver index of rats in the administration groups decreased significantly(P<0.01).The results of HE staining showed that the liver tissue structure of rats in the control group was complete and there was no obvious abnormality.The model group showed vacuolar degeneration and inflammatory infiltration of hepatocytes of rats.There was no obvious abnormality in the liver structure of rats in the administration groups.The results of blood lipid

关 键 词：2型糖尿病(T2DM) 肝脏脂质代谢 桑叶总黄酮 过氧化物酶增殖物激活受体-α(PPAR-α) 肉毒碱棕榈酰基转移酶-1(CPT-1) 

分 类 号：R22[医药卫生—中医基础理论] R242[医药卫生—中医学] R2-031R285.5