参芪降糖胶囊通过调节NF-κB/NLRP3通路对糖尿病动脉粥样硬化大鼠糖脂代谢、内皮细胞功能的影响

Impacts of Shenqi Jiangtang Capsules on glucose and lipid metabolism and endothelial cell function in diabetes atherosclerotic rats by regulating the NF-κB/NLRP3 pathway

关键词:  参芪降糖胶囊  糖尿病动脉粥样硬化  核因子-κB/NOD样受体蛋白3通路  糖脂代谢  内皮细胞

Key words:  Shenqi Jiangtang Capsules  atherosclerosis in diabetes  nuclear factor-κB/NOD like receptor protein 3 pathway  glucose and lipid metabolism  endothelial cell

摘要:

[目的] 探讨参芪降糖胶囊（简称SJC）对糖尿病动脉粥样硬化（AS）大鼠糖脂代谢、内皮细胞功能的影响及对核因子（NF）-κB/NOD样受体蛋白3（NLRP3）信号通路的调节作用。[方法] 建立AS大鼠模型，所有大鼠随机分为空白组、模型组、SJC低剂量组、SJC高剂量组和SJC高剂量+NLRP3激活剂组，观察各组大鼠一般情况，试剂盒检测血清空腹血糖（FBG）、空腹胰岛素（FINS）、胰岛素抵抗指数（HOMA-IR）、白介素（IL）-1β、IL-6、肿瘤坏死因子（TNF）-α含量，自动分析仪检测血清总胆固醇（TC）、三酰甘油（TG）、低密度脂蛋白胆固醇（LDL-C）和高密度脂蛋白胆固醇（HDL-C）的含量，苏木精-伊红（HE）染色观察主动脉病理学特征；敏感肌张力描计检测动脉血管内皮功能；免疫蛋白印迹（Western blot）技术验证磷酸化（p）-NF-κB p65、NF-κB p65、NLRP3蛋白表达；免疫组化检测主动脉半胱天冬酶-1（Cleaved-Caspase-1）、凋亡相关斑点样蛋白（ASC）蛋白表达。[结果] 与空白组对比，模型组大鼠体型瘦弱，毛发干枯，多食多饮多尿，精神恍惚，主动脉管腔内膜不规则增厚且形成斑块，炎症细胞浸润，平滑肌排列不规则，血清FBG、FINS、HOMA-IR、TC、TG、LDL-C、IL-1β、IL-6、TNF-α及主动脉内膜p-NF-κB p65/NF-κB p65、NLRP3、Cleaved-Caspase-1、ASC蛋白表达水平升高，血管内皮最大舒张率和HDL-C水平下降（P＜0.05）；与模型组比较，SJC低、高剂量组大鼠毛色光泽增加，精神状态和多饮多尿状况好转，血管内斑块减少，中膜钙化少见，炎性细胞浸润减少，整体病变程度减轻，血清FBG、FINS、HOMA-IR、TC、TG、LDL-C、IL-1β、IL-6、TNF-α及主动脉内膜p-NF-κB p65/NF-κB p65、NLRP3、Cleaved-Caspase-1、ASC蛋白表达水平降低，血管内皮最大舒张率和HDL-C水平升高（P＜0.05）；与SJC高剂量组对比，SJC高剂量+NLRP3激活剂组大鼠DM体征和血管病变程度加重，血清FBG、FINS、HOMA-IR、TC、TG、LDL-C、IL-1β、IL-6、TNF-α及主动脉内膜p-NF-κB p65/NF-κB p65、NLRP3、Cleaved-Caspase-1、ASC蛋白表达水平增加，血管内皮最大舒张率和HDL-C水平降低（P＜0.05）。[结论] SJC可能通过抑制NF-κB/NLRP3信号通路改善AS大鼠糖脂代谢和内皮细胞功能。

Abstract:

[Objective] To investigate the impacts of Shengi Jiangtang Capsules(SJC) on glucose and lipid metabolism, endothelial cell function in diabetes atherosclerosis(AS) rats,and its regulatory effect on the nuclear factor(NF)-κB/NOD like receptor protein 3(NLRP3) signaling pathway. [Methods] An AS rat model was established,and all rats were randomly grouped into blank group,model group,SJC low-dose group,SJC high-dose group,and SJC high-dose+NLRP3 activator group,the general situation of rats in each group was observed,the kit was applied to detect the levels of serum fasting blood glucose(FBG),fasting insulin(FINS),insulin resistance index(HOMA-IR),interleukin-1β(IL-1β),IL-6,and tumor necrosis factor-α(TNF-α),the automatic analyzer was applied to detect the contents of serum total cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C) and high-density lipoprotein cholesterol(HDL-C),HE staining was applied to observe the pathological characteristics of the aorta;the sensitive tonometer was applied to detect the endothelial function of arteries;Western blot was applied to verify the protein expression of phosphorylated(p)-NF-κB p65,NF-κB p65,NLRP3;the expressions of Cleaved-Caspase-1(Cleaved-Caspase-1) and apoptosis-related speck-like protein(ASC) in aorta were detected by immuno- histochemistry. [Results] Compared with the blank group,the rats in the model group had a lean and weak body shape,dry hair,excessive eating,drinking and urination,confusion,irregular thickening and formation of plaques in the aortic lumen,infiltration of inflammatory cells,and irregular arrangement of smooth muscle,the levels of serum FBG,FINS,HOMA-IR,TC,TG,LDL-C,IL-1β,IL-6,TNF-α,and the protein expression levels of p-NF-κB p65/NF-κB p65,NLRP3,Cleaved-Caspase-1,and ASC in the aortic intima increased,the maximum diastolic rate of vascular endothelium and HDL-C level decreased(P＜0.05);compared with the model group,the fur color and luster of the rats in the low-dose and high-dose SJC groups were improved,their mental state and polydipsia and polyuria improved,intravascular plaques decreased,medial calcification was rare,inflammatory cell infiltration decreased,and the overall severity of the disease decreased,the levels of serum FBG,FINS,HOMA-IR,TC,TG,LDL-C,IL-1β,IL-6,TNF-α,and the protein expression levels of p-NF-κB p65/NF-κB p65,NLRP3,Cleaved-Caspase-1,and ASC in the aortic intima decreased,the maximum diastolic rate of vascular endothelium and HDL-C level increased(P＜0.05);compared with the SJC high-dose group,the signs of DM and the degree of vascular lesions of rats in the SJC high-dose+NLRP3 activator group worsened,the levels of serum FBG,FINS,HOMA-IR,TC,TG,LDL-C,IL-1β,IL-6,TNF-α,and the protein expression levels of p-NF-κB p65/NF-κB p65,NLRP3,Cleaved-Caspase-1,and ASC in the aortic intima increased,the maximum diastolic rate of vascular endothelium and HDL-C level decreased(P＜0.05). [Conclusion] SJC may improve glucose and lipid metabolism and endothelial cell function in AS rats by inhibiting the NF-κB/NLRP3 signaling pathway.