白芍总苷对动脉粥样硬化大鼠TNF-α/P38MAPK/NF-κB/RBP4信号通路及其血清IL-17,IL-27,IL-33影响的研究

Total Glucosides of Paeony Has Effect on TNF-α/P38MAPK/NF-κB/RBP4 Signaling Pathways in Rats with Atherosclerosis and Its Regulating Effect on IL-7, IL-27 and IL-33 in Serum

目的:观察白芍总苷(Total glucosides of paeony,TGP)对动脉粥样硬化(Atherosclerosis,AS)大鼠TNF-α/P38MAPK/NF-κB/RBP4信号通路的影响及对血清中白细胞介素-17 (Interleukin-17,IL-17),白细胞介素-27 (Interleukin-27,IL-27),白细胞介素-33 (Interleukin-33,IL-33)的调节作用。方法:30只雄性SD大鼠随机分成正常组、模型组和TGP组,每组10只。除正常组外,模型组和TGP组大鼠均通过喂养高脂饲料结合腹腔注射维生素D3的方法建立大鼠AS模型,共12周,同时TGP组大鼠按照200 mg/(kg·d)剂量在造模过程中同步连续灌胃给药12周,正常组和模型组则同步给予等体积生理盐水。以上各组大鼠于建立模型第12周末处死,称取各组大鼠体质量及心脏组织质量计算心脏指数(Cardiac index,CI);HE染色观察各组大鼠主动脉形态结构病理学变化;采用ELISA试剂盒测定各组大鼠血清中IL-27、IL-17、IL-33的水平;采用蛋白印记法检测大鼠主动脉中肿瘤坏死因子-α(Tumor necrosis factor-α,TNF-α)、p38丝裂原活化蛋白激酶(p38 Mitogen activated protein kinase,p38MAPK)、核因子κB (Nuclear factor kappa-B,NF-κB)、视黄醇结合蛋白4 (Retinol-binding protein 4,RBP4)的表达。结果:与正常组比较,模型组大鼠的CI、主动脉HE染色病理评分、细胞因子水平(IL-27、IL-17、IL-33)以及TNF-α、p38MAPK、NF-κB、RBP4蛋白相对表达量均显著增加(P <0.05);与模型组比较,TGP组大鼠的CI、主动脉HE染色病理评分、细胞因子水平(IL-27、IL-17、IL-33)以及TNF-α、p38MAPK、NF-κB、RBP4蛋白相对表达量均显著降低(P <0.05)。结论:TGP对AS大鼠心脏具有显著的保护作用,可改善其主动脉粥样硬化程度,同时抑制TNF-α/P38MAPK/NF-κB/RBP4信号通路相关蛋白的表达,减少IL-17、IL-27及IL-33等炎症细胞因子的表达。

Objective: To observe the effect of total glucosides of paeony(TGP) on TNF-α/P38 MAPK/NF-κB/RBP4 signaling pathways in rats with atherosclerosis(AS) and its regulating effect on interleukin-17(IL-17), interleukin-27(IL-27)and interleukin-33(IL-33) in serum. Methods:A total of 30 male SD rats were randomly divided into the normal group,the model group and the TGP group,10 rats in each group. Except those in the normal group,the rats in the model group and the TGP group were fed with high-fat diet and given intraperitoneal injection of vitamin D3 for 12 weeks to establish the rat models of AS;the rats in the TGP group were synchronously given 200 mg/(kg·d) of total glucosides of paeony by gavage for12 weeks during the modeling,while the normal group and the model group were given with the same volume of saline. The rats in the above groups were sacrificed at the end of the 12 thweek after the modeling;the cardiac index(CI) was calculated by weighting the body and the heart of rats in each group;the pathological changes in morphology and structure of aortae in rats in each group were observed after HE staining;the levels of IL-27,IL-17 and IL-33 in serum in rats in each group were detected through ELISA kits;the expressions of tumor necrosis factor-α(TNF-α), p38 mitogen activated protein kinase(p38 MAPK),nuclear factor kappa-B(NF-κB) and retinol-binding protein 4(RBP4) in the aortae of rats were detected through the western blot. Results:Compared with those in the normal group,the score of CI,the pathological score of aortae after HE staining,the levels of IL-27,IL-17 and IL-33,and the relative protein expressions of TNF-α,p38 MAPK,NF-κB and RBP4 in rats in the model group were significantly increased(P < 0.05);compared with those in the model group,the above indexes in rats in the TGP group were significantly decreased(P < 0.05). Conclusion:TGP has significantly protective effect on the heart of AS rats, which can reduce the degree of aortic atherosclerosis, inhibit the expressions of TNF-α/P38 MAPK/NF-κB/RBP4 signaling pathways-related proteins, and reduce the expressions of the inflammatory cytokines such as IL-17,IL-27 and IL-33.