麻黄拆分组分对风水水肿大鼠的干预机制研究

Study on the Intervention Mechanism of Ephedrae Herba Split Components on Fengshui Edema Rats

目的:探讨麻黄拆分组分干预风水水肿模型大鼠的作用机制。方法:随机选择60只雄性SD大鼠通过尾静脉注射阿霉素结合吹风刺激建立风水水肿模型,当大鼠出现四肢浮肿、恶风、弓背、毛松、尿量减少等症状即为造模成功,将造模成功的大鼠随机分为模型组、通宣理肺丸(4326 mg/kg)组、麻黄水煎液(4670 mg/kg)组及麻黄各拆分组分组:生物碱(51.48 mg/kg)组、非生物碱(215.28 mg/kg)组、多糖(276.12 mg/kg)组。连续灌胃干预21 d,正常对照组和模型组给予等体积蒸馏水。给药结束后,观察大鼠基础特征;试剂盒检测大鼠血清CRE、BUN水平;Flow sight检测大鼠肾组织中ROS及细胞凋亡、自噬水平;免疫荧光检测大鼠肾小管中自噬相关蛋白表达;Western Blot检测大鼠肾组织中凋亡、炎症及自噬相关蛋白表达;电镜观察大鼠肾组织中足细胞的超微结构。结果:麻黄生物碱和多糖组分可改善风水水肿大鼠四肢浮肿、恶风、弓背、毛松等症状,显著降低风水水肿大鼠血清CRE、BUN及肾脏ROS水平和肾组织细胞的凋亡、自噬,抑制ERK、PI3K、AKT、mTOR磷酸化,调节凋亡、炎症、自噬相关蛋白表达,麻黄生物碱组分可显著改善风水水肿模型大鼠肾小球基底膜增厚,足突增宽、弥漫性融合及自噬体增多的现象。结论:麻黄生物碱组分和多糖组分可能通过阻断氧化应激抑制ERK-NLRP3炎性小体,抑制细胞凋亡,并通过激活PI3K/mTOR信号转导抑制肾脏细胞自噬,进而改善风水水肿模型大鼠的肾脏病理损伤。

Objective:To explore the action mechanism of Ephedrae Herba split components intervening in Fengshui edema model rats.Methods:60 SD male rats were used to establish Fengshui edema models by tail vein injection adriamycin combined with blowing stimulation.The model was successfully established when the rats showed symptoms such as swollen limbs,bowed back,loose hair and decreased urine output.They were randomly divided into model group,Tongxuanlifei pill(4326 mg/kg)group,ephedra(4670 mg/kg)group,ephedra split components group,including alkaloid component(51.48 mg/kg),nonalkaloid component(215.28 mg/kg),polysaccharide components(276.12 mg/kg).The corresponding drugs were administered by gavage for 21 days,and the normal control group and model group were given the same amount of distilled water.After administration,the basic characteristics of rats were observed,the levels of CRE and BUN in rat serum were detected by the kits.The ROS and apoptosis,autophagy levels in rat kidneys were detected by Flow sight;The expression levels of autophagy-related proteins in rat renal tubules were detected by immunofluorescence;The autophagy and inflammation-related proteins in rat kidneys were detected by Western Blot method and the ultrastructure of podocytes in rat kidney tissue was observed by electron microscopy.Results:The Ephedrae Herba alkaloid and polysaccharide components could improve the swollen limbs,bowed back,loose hair and other symptoms in rats with Fengshui edema,reduced the levels of CRE,BUN in th seurm,ROS in the kidney and apoptosis,autophagy in kidney tissue,inhibited ERK,PI3K,AKT,mTOR phosphorylation,apoptosis,inflammatory,autophagy-related protein expressions.The alkaloid components significantly improved the obvious thickening of the glomerular basement membrane,the widening of the foot process,the diffuse fusion and the increase of autophagosomes in the Fengshui edema model rats.Conclusion:The Ephedrae Herba alkaloid and polysaccharide components may inhibit ERK-NLRP3 inflammasome by blocking oxidative stress,inhibit cell apoptosis,and inhibit excessive autophagy in kidney cells by activating PI3K/mTOR signal transduction,thereby improving kidney pathological damage in Fengshui edema model rats.