摘要
目的:观察镇肝熄风汤与法尼醇受体(FXR)激动剂对自发性高血压大鼠(SHR)血压、血浆胆汁酸谱及FXR、成纤维细胞生长因子21(FGF21)的影响。方法:9周龄雄性SHR 60只随机分为模型组、贝那普利组(1.0 mg·kg^(-1)·d^(-1))、镇肝熄风汤高、中、低剂量组(34.5、17.25、8.625 g·kg^(-1)·d^(-1))、激动剂组(10 mg·kg^(-1)·d^(-1)),同源9周龄雄性Wistar-Kyoto(WKY)大鼠10只为空白组。空白组、模型组予等量蒸馏水灌胃,连续干预8周。采集大鼠肝脏与血浆,用动物无创血压计以尾套法测量各组大鼠血压,用超高效液相色谱-串联质谱法(UPLC-MS/MS)检测各组大鼠血浆胆汁酸谱;用酶联免疫吸附测定法(ELISA)检测血浆FGF21表达水平;用实时荧光定量聚合酶链反应(Real-time PCR)检测各组大鼠肝脏组织中FXR、FGF21基因表达水平;用蛋白免疫印迹法(Western blot)检测各组大鼠肝脏组织中FXR、FGF21蛋白表达水平。结果:镇肝熄风汤降低了SHR收缩压与舒张压。与空白组比较,模型组血浆熊去氧胆酸、猪去氧胆酸、去氧胆酸、石胆酸、甘氨鹅去氧胆酸、甘氨熊去氧胆酸、甘氨猪去氧胆酸表达明显降低(P<0.05,P<0.01);血浆FGF21表达显著降低(P<0.01);肝脏FXR、FGF21 mRNA表达显著降低(P<0.01)。与模型组比较,镇肝熄风汤高剂量组熊去氧胆酸、猪去氧胆酸、甘氨胆酸、甘氨鹅去氧胆酸、甘氨熊去氧胆酸、甘氨猪去氧胆酸表达明显降低(P<0.05,P<0.01);低、中剂量组去氧胆酸表达明显升高(P<0.05);低剂量组甘氨胆酸表达明显降低(P<0.05);激动剂组甘氨胆酸表达明显降低(P<0.05)。镇肝熄风汤各剂量组与激动剂组血浆FGF21表达显著升高(P<0.05,P<0.01);肝脏FXR、FGF21 mRNA表达显著升高(P<0.05,P<0.01)。镇肝熄风汤中剂量组、激动剂组肝脏FXR蛋白表达显著升高(P<0.05,P<0.01);镇肝熄风汤高剂量组肝脏FGF21表达显著升高(P<0.01)。结论:镇肝熄风汤的降压作用可能与调节SHR血浆胆汁酸谱结构,上调肝脏FXR-FGF21表达有关。
Objective:To observe the effects of Zhengan Xifengtang on the blood pressure,bile acid spectrum in plasma,and the expression of farnesoid X receptor(FXR)and fibroblast growth factor 21(FGF21)in spontaneously hypertensive rats(SHR).Methods:Sixty 9-week-old males of SHR were randomized into SHR,benazepril(1.0 mg·kg^(-1)·d^(-1)),high-,medium-,and low-dose(34.5,17.25,8.625 g·kg^(-1)·d^(-1))Zhengan Xifengtang,and agonist(10 mg·kg^(-1)·d^(-1))groups,and 10 male Wistar-Kyoto(WKY)rats of 9 weeks old were selected as the WKY group.The WKY and SHR groups were administrated with an equal volume of distilled water for 8 consecutive weeks.The liver and plasma samples were collected from the rats.The blood pressure of rats in each group was measured by a tail-cuff method with an animal non-invasive sphygmomanometer.The bile acid spectrum in the plasma of rats in each group was determined by ultra-high performance liquid chromatography-tandem mass spectrometry(UPLC-MS/MS).The expression of FGF21 in the plasma was measured by enzyme-linked immunosorbent assay.The mRNA and protein levels of FXR and FGF21 in the liver tissue were determined by real-time PCR and Western blot,respectively.Results:Zhengan Xifengtang reduced the systolic and diastolic blood pressure of SHR.Compared with the WKY group,the SHR group presented declined levels of ursodeoxycholic acid,hyodeoxycholic acid,deoxycholic acid,lithocholic acid,glycochenodeoxycholic acid,glycoursodeoxycholic acid,and glycohyodeoxycholic acid in the plasma(P<0.05,P<0.01),reduced expression of FGF21 in the plasma(P<0.01),and down-regulated mRNA levels of FXR and FGF21 in the liver(P<0.01).Compared with those in the SHR group,the levels of ursodeoxycholic acid,hyodeoxycholic acid,glycocholic acid,glycochenodeoxycholic acid,glycoursodeoxycholic acid,and glycohyodeoxycholic acid became lowered in the highdose Zhengan Xifengtang group(P<0.05,P<0.01).The level of deoxycholic acid declined in the low-and medium-dose Zhengan Xifengtang groups(P<0.05).The level of glycocholic acid reduced in the low-dose Zhengan Xifengtang group(P<0.05)and the agonist group(P<0.05).The expression of FGF21 in the plasma and the mRNA levels of FXR and FGF21 in the liver increased in the high-,medium-,and low-dose Zhengan Xifengtang groups and the agonist group(P<0.05,P<0.01).The protein level of FXR in the liver of the medium-dose group and the agonist group was up-regulated(P<0.05,P<0.01).The expression of FGF21 in the liver of the highdose group increased(P<0.01).Conclusion:Zhengan Xifengtang may exert the antihypertensive effect by regulating the bile acid spectrum in the plasma and up-regulating the expression of FXR-FGF21 in the liver of SHR.
作者
凌必时
金华
祁尚文
刘双芳
司美龙
刘志军
LING Bishi;JIN Hua;QI Shangwen;LIU Shuangfang;SI Meilong;LIU Zhijun(Gansu University of Chinese Medicine,Key Laboratory of Dunhuang Medicine and Transformation Ministry of Education,Lanzhou 730000,China;Shenzhen Luohu People's Hospital,Shenzhen 518000,China;Affiliated Hospital of Gansu University of Chinese Medicine,Lanzhou 730000,China)
出处
《中国实验方剂学杂志》
北大核心
2025年第23期109-118,共10页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(82160886,81860832)
甘肃省自然科学基金项目(20JR10RA315)
甘肃省省级科技计划项目(23JRRA1524)
甘肃省省级重点人才项目(甘组通字[2023]20号)
甘肃省自然科学基金-优秀博士生项目(22JR5RA579)。
