镇肝熄风汤对自发性高血压大鼠胆汁酸-TGR5/Rho/ROCK/AgRP信号通路的影响  

Effects of Zhengan Xifeng decoction on bile acid-TGR5/RhO/ROCK/AgRP signaling pathway in spontaneously hypertensive rats

作　　者：祁尚文 金华 凌必时 刘双芳 司美龙 QI Shang-wen;JIN Hua;LING Bi-shi;LIU Shuang-fang;SI Mei-long(Clinical College of Chinese Medicine,Gansu University of Chinese Medicine,Lanzhou 730000,Gansu Province,China;Key Laboratory of Dunhuang Medicine,Ministry of Education,Gansu University of Chinese Medicine,Lanzhou 730000,Gansu Province,China)

机构地区：[1]甘肃中医药大学中医临床学院,甘肃兰州730000 [2]甘肃中医药大学敦煌医学与转化教育部重点实验室,甘肃兰州730000

出　　处：《中国临床药理学杂志》2024年第22期3290-3295,共6页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(82160886,81860832);甘肃省自然科学基金资助项目(20JR10RA315,22JR5RA579);甘肃省省级重点人才基金资助项目(甘组通字[2023]20号)。

摘　　要：目的观察镇肝熄风汤对自发性高血压大鼠(SHR)胆汁酸-武田G蛋白偶联受体5(TGR5)/Ras同源物(Rho)/Rho关联含卷曲螺旋结合蛋白激酶(ROCK)/刺鼠关联蛋白(AgRP)信号通路的影响。方法将70只SHR随机分为模型组、对照组(灌胃0.5mg·kg^(-1)·d^(-1)苯磺酸氨氯地平片)和低、中、高剂量实验组(分别灌胃8.630×10^(3),1.725×10^(4)、3.450×10^(4)mg·kg^(-1)·d^(-1)镇肝熄风汤),每组14只;另取15只Wistar-Kyoto大鼠为正常组。模型组和正常组均灌胃给予等量蒸馏水。6组大鼠每天给药1次,连续8周。用超高效液相色谱-串联质谱法检测各组大鼠空肠内容物和结肠内容物胆汁酸谱,用蛋白质印迹法检测下丘脑组织中TGR5、Rho、ROCK、AgRP蛋白的相对表达水平。结果正常组、模型组、对照组和低、中、高剂量实验组比较,空肠内容物胆汁酸谱显著改变,共发现6种差异胆汁酸,分别为甘氨熊去氧胆酸、甘氨猪去氧胆酸、猪去氧胆酸、熊去氧胆酸、牛磺鹅去氧胆酸和鹅去氧胆酸;结肠内容物胆汁酸谱显著改变,共发现4种差异胆汁酸,分别为去氧胆酸、熊去氧胆酸、石胆酸和猪去氧胆酸。正常组、模型组、对照组和中剂量实验组的TGR5蛋白相对表达水平分别为1.01±0.22、0.75±0.23、0.96±0.16和1.02±0.21,Rho蛋白相对表达水平分别为0.81±0.14、0.58±0.19、0.83±0.27和0.94±0.24,ROCK蛋白相对表达水平分别为0.99±0.27、0.65±0.13、1.07±0.22和1.06±0.24,AgRP蛋白相对表达水平分别为0.98±0.33、1.46±0.45、0.83±0.25和0.95±0.33。中剂量实验组和正常组的上述指标与模型组比较,在统计学上差异均有统计学意义(均P<0.05)。结论镇肝熄风汤可能通过胆汁酸-TGR5/Rho/ROCK/AgRP信号通路抑制AgRP表达,从而达到降低血压的目的。Objective To observe the effects of Zhengan Xifeng decoction on bile acid-Takeda G protein-coupled receptor 5(TGR5)/Ras homolog(Rho)/Rho-associated coiled-coil containing protein kinase(ROCK)/agouti-related protein(AgRP)signaling pathway in spontaneously hypertensive rats(SHR).MethodssSeventySHRwere randomly divided intothemodelgroup,control group(gavage administration of 0.5 mg·kg^(-1).d^(-1)aaml lodipine besylate tablets),experimental-L,-M,-H group(gavage administration of 8.630×10^(3),1.725×10^(4),3.450×10^(4)mg·kg^(-1)·d^(-1)Zhengan Xifeng decoction).Fifteen Wistar-Kyoto rats served as the normal group.The model and normal groups were given the same amount of distilled water.Six groups were treated for 8 weeks with once a day.The bile acid profiles of jejunum contents and colon contents were detected by ultra-high-performance liquid chromatography-tandem mass spectrometry,and the expression levels of TCR5,Rho,ROCK and AgRP proteins in the hypothalamus were detected by Western blot.Results Compared with the normal,mode,control groups and experimental-L,-M,-H groups,the bile acid spectrum of jejunum contents was significantly changed,and 6 different bile acids were found,namely glycoursodeoxycholic acid,glycohyodeoxycholic acid,hyodeoxycholic acid,ursodeoxycholic acid,taurochenodeoxycholic acid and chenodeoxycholic acid.The bile acid spectrum of colon contents was significantly changed,and 4 different bile acids were found,namely deoxycholic acid,ursodeoxycholic acid,lithocholic acid and hyodeoxycholic acid.The relative expression levels of TGR5 protein in normal,model,control and experimental-M groups were 1.01±0.22,0.75±0.23,0.96±0.16 and 1.02±0.21;the relative expression levels of Rho protein were 0.81±0.14,0.58±0.19,0.83±0.27 and 0.94±0.24;the relative expression levels of R0CK protein were 0.99±0.27,0.65±0.13,1.07±0.22 and 1.06±0.24;the relative expression levels of AgRP protein were 0.98±0.33,1.46±0.45,0.83±0.25 and 0.95±0.33,respectively.Compared with the model group,the above index

关 键 词：镇肝熄风汤 高血压 胆汁酸 武田G蛋白偶联受体5 刺鼠关联蛋白 

分 类 号：R28[医药卫生—中药学]