当归补血汤及其活性成分通过抑制NLRP3炎症小体活化改善糖尿病肾病的实验研究

Danggui Buxue Decoction and Its Active Ingredients Improv Diabetic Nephropathy by Inhibiting the Activation of NLRP3 Inflammasome

目的:探究不同剂量的当归补血汤及其活性成分黄芪甲苷改善糖尿病肾病的作用机制及与NOD样受体蛋白3(nod-like receptor protein 3,NLRP3)炎症小体的相关性。方法:5 w~8 w的雄性GK大鼠分为模型对照组、阳性药格列喹酮10 mg/kg、盐酸贝那普利片10 mg/kg、当归补血汤2、4、8 g/kg组、黄芪甲苷40 mg/kg组,每组10只;10只雄性Wistar大鼠作为正常对照组。各组灌胃相应药物,正常对照组和模型对照组灌胃生理盐水,连续8 w。检测随机血糖、空腹血糖;计算肾脏系数;Elisa法检测大鼠尿液中白蛋白含量;HE染色观察肾脏组织病理改变;Western Blot法检测大鼠肾脏NLRP3、ASC、Caspase-1蛋白的表达。结果:与正常对照组相比,模型对照组大鼠的血糖、肾脏系数、尿白蛋白含量、炎症蛋白表达量均显著升高(P<0.01);与模型对照组比较,当归补血汤4、8 g/kg及其活性成分黄芪甲苷0.04 g/kg均可改善GK大鼠的体质量、血糖、肾脏肥大的情况,显著降低尿白蛋白含量,能有效抑制NLRP3小体的活化,其中8 g/kg组对调节血糖、降低尿白蛋白含量效果最为显著(P<0.01)。结论:不同剂量当归补血汤及其活性成分黄芪甲苷对GK大鼠肾脏均具有一定的保护作用,其机制与抑制NLRP3炎症小体密切相关。

Objective:To explore the mechanism of Danggui Buxue Decoction(DBD)of different doses and its active ingredient astragaloside IV in improving diabetic nephropathy and its correlation with nod-like receptor protein 3(NLRP3)inflammasome.Methods:Male GK rats aged 5-8 weeks were divided into a model group,a positive drug gliquidone(10 mg/kg)group,a benazepril hydrochloride tablets(10 mg/kg)group,low-,medium-,and high-dose DBD groups(2 g/kg,4 g/kg,and 8 g/kg),and an astragaloside IV(40 mg/kg)group,with 10 rats in each group.Ten male Wistar rats were assigned to the control group.Rats were treated with corresponding drugs or normal saline by gavage for 8 weeks.Random blood glucose and fasting blood glucose were detected,and the renal index was calculated.ELISA was used to determine the urinary albumin content.HE staining was used to observe pathological changes of renal tissues.The protein expression of NLRP3,ASC,and Caspase-1 in the kidney was detected by Western blot.Results:Compared with the control group,the model group showed increased blood glucose content,renal index,urinary albumin,and inflammatory protein expression(P<0.01).Compared with the model group,the medium-,and high-dose DBD groups and the astragaloside IV group showed improved body weight,blood glucose,and renal hypertrophy of GK rats,reduced content of urinary albumin,and inhibited activation of NLRP3 inflammasome,while the high-dose DBD group displayed the most significant effect in regulating blood glucose and reducing urinary albumin(P<0.01).Conclusion:DBD of different doses and its active ingredient astragaloside IV can protect the kidney of GK rats,and the mechanism is closely related to the inhibition of NLRP3 inflammasome.