利胆软坚方降脂减肥作用及其对胆汁酸代谢轮廓谱的影响

Effect of Lidan Ruanjian Prescription on Lipid-lowering and Antiobesity and Metabolic Profile of Bile Acid

目的:基于肥胖“从胆论治”的学术观点,研究利胆软坚方对高脂饮食诱导肥胖大鼠的降脂减肥作用及其对血清胆汁酸轮廓谱的影响,探讨其作用机制。方法:42只大鼠高脂饲料喂养9周建立肥胖大鼠模型,取24只造模成功大鼠随机分为模型组、利胆软坚方高、低剂量组(30,15 g·kg^-1),每组8只,另取8只正常大鼠作为正常组,模型组和正常组给予生理盐水,给药组给予相应剂量药物,灌胃4周。测定大鼠体质量、肝脏质量、脂肪质量等肥胖指标;胆管插管术引流监测2 h内胆汁流量;全自动生化仪测定血清总胆固醇(TC),甘油三酯(TG),高密度脂蛋白胆固醇(HDL-C),低密度脂蛋白胆固醇(LDL-C)水平;超高效液相色谱-质谱联用(UPLC-MS/MS)法测定大鼠血清胆汁酸代谢轮廓谱中各胆汁酸的含量。结果:与正常组比较,模型组体质量、肝脏质量、脂肪质量均显著升高(P<0.01),血清TC,TG,LDL-C水平明显上升(P<0.05,P<0.01),2 h内胆汁总分泌量及各测试点胆汁流量均降低,初级胆汁酸占比明显下降(P<0.05),血清总胆汁酸含量显著降低(P<0.01),血清胆汁酸轮廓谱中胆酸(CA),脱氧胆酸(DCA),鹅去氧胆酸(CDCA),猪去氧胆酸(HDCA),牛磺胆酸(TCA),牛磺去氧胆酸(TDCA),牛磺鹅去氧胆酸(TCDCA),牛磺猪去氧胆酸(THDCA),甘氨去氧胆酸(GDCA)含量明显降低(P<0.05,P<0.01);与模型组比较,利胆软坚方高、低剂量组体质量、肝脏质量明显降低(P<0.05,P<0.01),利胆软坚方高剂量组血清TC,TG,LDL-C水平明显下降(P<0.05,P<0.01),低剂量组TG水平明显下降(P<0.05),高剂量组在给药后1~1.5 h胆汁流量明显增加(P<0.05),高、低剂量组初级胆汁酸占比明显升高(P<0.05),利胆软坚方高剂量组TCA,DCA,甘氨胆酸(GCA),GDCA水平明显升高(P<0.05,P<0.01),低剂量组仅TCA,TCDCA水平明显升高(P<0.05)。结论:利胆软坚方具有降脂减肥作用,其机制可能与其增加胆汁分泌量,增加初级胆汁酸合成及调节胆汁酸轮廓谱有关。

Objective:To investigate the pharmacodynamics effects of antiobesity,lipid-lowering and the regulations of serum bile acid profiles of Lidan Ruanjian prescription(LDRJ)in obesity rats induced by highfat diet.Method:The 42 rats were fed high-fat diet for 9 weeks to establish model of obese rats,24 rats were randomly divided into model group,high and low-dose LDRJ group(30,15 g·kg^-1).Another 8 normal rats were selected as the normal group.The model group and normal group were given normal saline,and drug group was given the corresponding dose of drug for 4 weeks.Body weight,liver weight,white adipose tissue(WAT)weight were determined after administration medicine for 4 weeks.The bile flow of the rats was measured by bile duct intubation and fasting serum lipid levels of total cholesterol(TC),total triglyceride(TG),highdensity lipoprotein cholesterol(HDL-C),low-density lipoprotein cholesterol(LDL-C)were detected by automatic biochemical analyzer.Ultra-performance liquid chromatography-tandem mass spectrometry(UPLCMS/MS)assay was used to test serum bile acid profile of each group rats.Result:Compared with the control group,the average body weight,liver weight,WAT weight of the model group were significantly increased(P<0.01),while the fasting serum TC,TG and LDL-C levels were elevated(P<0.05,P<0.01).The total bile secretion and bile flow at each test point within 2 h were decreased and the proportion of primary bile acids was decreased(P<0.05).The serum total bile acid content decreased significantly(P<0.01),levels of cholic acid(CA),deoxycholic acid(DCA),chenodeoxycholic acid(CDCA),hyodeoxycholic acid(HDCA),taurocholic acid(TCA),taurodeoxycholic acid(TDCA),taurochenodeoxycholic acid(TCDCA),taurohyodeoxycholic acid(THDCA)and glycodeoxycholic acid(GDCA)were significantly reduced(P<0.05,P<0.01).Compare with model group,body weight,liver weight in high and low-dose LDRJ groups reduced significantly(P<0.05,P<0.01).Fasting serum TC,TG and LDL-C levels were decreased in high-dose group(P<0.05,P<0.01),so did as TG levels in low-dose group(P<0.05).The bile flow rate increased significantly in high-dose group 1~1.5 h after administration(P<0.05).All dose treatment groups increased the proportion of primary bile acids(P<0.05)and changed the bile acid profile,especially elevated the bile acid levels of TCA,DCA,glycocholic acid(GCA),GDCA in high-dose LDRJ group(P<0.05,P<0.01),while TCA and TCDCA in low-dose group(P<0.05).Conclusion:LDRJ has significant lipid-lowering and antiobesity effects and the mechanism might involve the increase of bile secretion,the stimulation of primary bile acid synthesis and the regulation of bile acid profile.