葛根芩连汤干预2型糖尿病前期大鼠的代谢机制研究

Study on the Mechanism of Gegen Qin Lian Decoction in Preventing Type 2 Diabetic Rats

目的探讨葛根芩连汤对高脂诱导的2型糖尿病大鼠预防作用的代谢机制。方法将SD大鼠随机分为正常组(8只)和高脂造模组(40只),分别使用基础饲料和高脂饲料喂养12周。高脂造模组大鼠产生胰岛素抵抗(IR)后,将高脂造模组大鼠再随机分为模型组及葛根芩连汤低、中、高剂量组(1.65、4.96、14.86 g·kg-1),每组8只。模型组及不同剂量给药组继续给予高脂饲料喂养,给药组给予不同剂量的葛根芩连汤进行干预,灌胃给药,给药体积为10 mL·kg-1,每天1次,连续16周。检测大鼠空腹血糖值及空腹胰岛素含量,计算IR指数;使用超高效液相色谱-四级杆飞行时间串联质谱(UHPLC/Q-TOF-MS)结合多元分析方法,分析大鼠血清中的内源性代谢物及其变化差异,寻找葛根芩连汤干预IR大鼠及预防2型糖尿病的潜在生物标记物。结果高脂饲料喂养12周后,与正常组比较,造模组大鼠的IR指数明显升高(P<0.05),血糖无明显变化(P>0.05)。给药16周后,与正常组比较,模型组大鼠的IR指数及血糖值均显著升高(P<0.05,P<0.01);与模型组比较,葛根芩连汤低、中剂量组大鼠的IR指数显著降低(P<0.01),葛根芩连汤高剂量组的IR指数无明显变化(P>0.05);葛根芩连汤低剂量组的空腹血糖值明显降低(P<0.05),而葛根芩连汤中、高剂量组的空腹血糖值无明显变化(P>0.05)。大鼠血清代谢组分析鉴定共得到4种潜在代谢标志物,分别是Hippuric acid(马尿酸)、12-Ketodeoxycholic acid(12-去氧胆酸)、3-Oxo-4,6-choladienoic acid和LysoPC[20∶4(5Z,8Z,11Z,14Z)](溶血磷脂酰胆碱,LPC)。结论葛根芩连汤可以降低IR大鼠的胰岛素抵抗指数,同时可预防血糖升高,其机制可能与调节胆汁酸代谢、磷脂代谢以及肠道菌群代谢有关。

Objective To investigate the metabolic mechanism of Gegen Qin Lian Decoction(GQD)on prevention of type 2 diabete in rats induced by high-fat diet.Methods SD rats were randomly divided into normal group and high-fat group,fed with basic diet and high-fat diet respectively.After IR was produced in high-fat group,highfat group rats were randomly divided into model group,low-dose group(GQD-L,1.65 g·kg-1),medium dose group(GQD-M,4.96 g·kg-1)and high-dose group(GQD-H,14.86 g·kg-1).The model group continued to be fed with high-fat diet,while the different dose groups were given different doses of GQD for intervention.After 16 weeks of GQD administration,the blood glucose and IR indexes of all rats were detected,and the endogenous metabolites and their differences in serum samples were analyzed by Ultra-high Performance Liquid Chromatography-quadrupole Time-of-flight Mass Spectrometry(UHPLC/Q-TOF-MS)and multivariate analysis,in order to find the potential biomarkers for GQD intervening IR rats and preventing obese type 2 diabetes.Results After 12 weeks of high fat diet,compared with the normal group,the IR index of the model group was significantly higher(P<0.05),and there was no difference in blood glucose(P>0.05).After 16 weeks of GQD administration,compared with the normal group,the IR index of the model group was significantly increased(P<0.01),and the blood glucose was significantly increased(P<0.05).Compared with the model group,the IR indexes of the GQD-L group and the GQD-M group were significantly reduced(P<0.05),and there was no difference between the model group and the GQD-H group(P>0.05).The blood glu cose of GQD-L group decreased significantly(P<0.05),but there was no difference before and after treatment in GQD-M group and GQD-H group(P>0.05).Four potential metabonomic biomarkers of treating IR in rats with GQD were identified by the metabonomic study of serum samples,including hippuric acid,12 ketodeoxylic acid,3-oxo-4,6-choledienoic acid,and lysoPC[20:4(5 Z,8 Z,11 Z,14 Z)].Conclusion GQD can reduce the IR index of IR rats and prevent the increase of blood sugar simultaneously.The mechanism may be related to intestinal microecology metabolism,bile acid metabolism and phospholipid metabolism.