摘要
目的:观察温胆汤对肥胖痰湿证大鼠糖脂代谢、磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)信号通路关键分子蛋白表达的影响,探讨温胆汤干预肥胖痰湿证改善糖脂代谢的内在机制。方法:将126只大鼠随机分成2组:空白组16只,造模组110只,空白组采用基础饲料喂养,造模组采用高脂饲料喂养,共8周,制作肥胖痰湿证动物模型。造模成功后,视体质量情况,按序淘汰体质量过轻者,遴选出40只肥胖大鼠,随机分为模型对照(MC)组、奥利司他(ORLI)组、温胆汤低(WDD-L)、中(WDD-M)、高剂量(WDD-H)组,每组8只;另在空白组大鼠中随机选取8只设为正常对照(NC)组。ORLI组(32.40 mg/kg)、WDD-L组、WDD-M组和WDD-H组[按4.45、8.90、17.80 g/kg剂量(以生药量计算)]给予灌胃,MC和NC组则均给予等量蒸馏水灌胃,每天1次,共6周。治疗结束后,检测并计算大鼠体质量、Lee’s指数、脂体比和肥胖率,采用全自动生化分析仪检测大鼠血总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)及空腹血糖(FBG)水平;采用酶联免疫吸附测定法(ELISA)检测空腹胰岛素(FINS),并计算稳态模型胰岛素抵抗指数(HOMA-IR);采用Western Blot检测腹周脂肪细胞PI3K/Akt信号通路关键分子PI3K、PIP3、p-Akt、Akt蛋白表达。结果:与空白组比较,造模组体质量和Lee’s指数显著升高(P<0.01),肥胖率>20%,有形体肥胖,活动度降低,食欲降低,皮毛无光泽、蓬松,便溏,对外界反应能力降低,饮水量减少等痰湿证型症状,说明高脂饲料喂养可成功复制肥胖痰湿证大鼠模型。与NC组比较,MC组大鼠体质量、Lee’s指数、脂体比显著升高(P<0.05,P<0.01),血脂、FBG、FINS及HOMA-IR水平均显著改变(P<0.01),腹周脂肪细胞PI3K、PIP3、p-Akt、Akt蛋白表达显著降低(P<0.01)。与MC组比较,各用药组大鼠肥胖痰湿证型症状显著改善,大鼠体质量、脂体比显著降低(P<0.05,P<0.01),血脂、FBG、FINS及HOMA-IR水平显著改善(P<0.05,P<0.01),腹周脂肪细胞PI3K、PIP3、p-Akt、Akt蛋白表达显著升高(P<0.05,P<0.01)。与ORLI组比较,WDD-L、WDD-M组HDL-C水平显著降低(P<0.01)。结论:温胆汤能减轻肥胖大鼠体重,改善其痰湿证型症状,调节糖脂代谢,降低胰岛素水平,有效干预胰岛素抵抗,其作用机制可能是通过激活PI3K/Akt信号通路来实现的。
Objective:To observe the effects of Wendan Decoction on glucose and lipid metabolism and the expression of key molecules of phosphatidylinositol-3-kinase-protein kinase B(PI3K/Akt)signaling pathway in obese rats with phlegm dampness syndrome,and to explore the mechanism underlying the intervention of Wendan Decoction in obese rats with phlegm-dampness syndrome to improve glucose and lipid metabolism.Methods:A total of 126 rats were randomly divided into 2 groups(110 rats in the modeling group and 16 rats in the blank group),and the blank group was fed with basal diet while the modeling group was fed with high-fat diet for 8 weeks to create an animal model of obesity with phlegm-dampness syndrome.After successful modeling,the rats with too light weight were eliminated in order according to their body weight,40 obese rats were selected according to their body mass,and randomly divided into model control(MC)group,orlistat(ORLI)group,Wendan Decoction low dose(WDD-L)group,Wendan Decoction medium dose(WDD-M)group,and Wendan Decoction high dose(WDD-H)group,8 rats in each group;and 8 rats were randomly selected from the blank group to be set as the normal control(NC)group.ORLI group(32.40 mg/kg),WDD-L group(4.45 g/kg),WDD-M group(8.90 g/kg)and WDD-H group(17.80 g/kg)were given gavage by dose(in raw drug quantity),respectively,and equal amounts of distilled water were given to both MC and NC groups once daily for 6 weeks.After the treatment,samples were taken after anesthesia to detect and calculate body mass,Lee's index,lipid-body ratio and obesity rate,and blood total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),high-density lipoprotein cholesterol(HDL-C)and fasting blood glucose(FBG)levels were measured by automatic biochemical analyzer;Fasting insulin(FINS)was measured by enzyme-linked immunosorbent assay(ELISA)and the steady-state model insulin resistance index(HOMA-IR)was calculated;Western Blot was used to detect the expression of PI3K,PIP3,p-Akt and Akt proteins,key molecules of PI3K/Akt signaling pathway in peritoneal adipocytes.Results:Compared with the blank group,the body mass and Lee’s index of the modeling group were significantly higher(P<0.01),the obesity rate was>20%,and there were phlegm-damp symptoms such as physical obesity,decreased mobility,decreased appetite,lusterless and tight fur,loose stools,decreased responsiveness to the outside world,and decreased water intake,indicating that high-fat diet feeding can successfully replicate the obesity phlegm-damp syndrome rat model.Compared with the NC group,body mass,Lee’s index and lipid-to-body ratio were significantly higher in the MC group(P<0.05,P<0.01),blood lipid,FBG,FINS and HOMA-IR levels were significantly altered(P<0.01),and PI3K,PIP3,p-Akt and Akt protein expression in peritoneal adipocytes were significantly decreased(P<0.01).Compared with the MC group,the obesity and phlegm symptoms of rats in each dosing group were significantly improved,the body mass and lipid-to-body ratio of rats were significantly decreased(P<0.05,P<0.01),the blood lipid,FBG,FINS and HOMA-IR levels were significantly improved(P<0.05,P<0.01),and the expression of PI3K,PIP3,p-Akt and Akt proteins in peritoneal adipocytes were significantly increased(P<0.05,P<0.01).Compared with the ORLI group,the WDD-L,WDD-M group showed significantly lower HDL-C levels(P<0.01).Conclusion:Wendan Decoction can reduce the body weight of obese rats,improve their phlegm-damp symptoms,regulate glucolipid metabolism,reduce insulin levels,and effectively intervene in insulin resistance,and its mechanism of action may be achieved by activating PI3K/Akt signaling pathway.
作者
喻松仁
刘彩玲
钟友宝
熊乃鑫
徐佳玲
刘春燕
王萍
张洁
程绍民
YU Songren;LIU Cailing;ZHONG Youbao;XIONG Naixin;XU Jialing;LIU Chunyan;WANG Ping;ZHANG Jie;CHENG Shaomin(Experimental Animal Science and Technology Center,Jiangxi University of Chinese Medicine,Nanchang 330004,China;Xinjian District Hospital of Traditional Chinese Medicine,Nanchang 330100,China;Traditional ChineseMedicine School,Jiangxi University of Chinese Medicine,Nanchang 330004,China)
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2024年第10期5468-5473,共6页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
江西省自然科学基金项目(No.20212BAB206010)
江西省教育厅科学技术研究项目(No.GJJ201212)
国家自然科学基金项目(No.81960851)
江西省中医药管理局中医证候基础重点研究室(No.赣中医药科教字[2022]8号)
江西中医药大学中医体质-状态辨识健康管理研究团队(No.CXTD22016)。
