基于NLRP3/caspase-1/GSDMD通路研究黄精改善糖尿病大血管焦亡损伤的作用机制

Effect of Polygonati Rhizoma in improving pyroptosis injury of diabetic macroangiopathy via NLRP3/caspase-1/GSDMD pathway

探讨黄精调节NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)/天冬氨酸特异性半胱氨酸蛋白酶-1(cysteinyl aspartate specific proteinase-1,caspase-1)/消皮素D(gasdermin D,GSDMD)通路对糖尿病大血管焦亡损伤的影响。采用小剂量链脲佐菌素(streptozotocin,STZ)腹腔注射联合高糖高脂饮食诱导糖尿病大鼠模型。利用血糖检测仪、全自动生化分析仪、苏木素-伊红(hematoxylin-eosin,HE)染色、酶联免疫吸附测定(enzymelinked immunosorbent assay,ELISA)、免疫荧光、免疫组织化学及Western blot等方法,检测大鼠血糖含量、血脂水平、血管厚度、炎症因子含量及焦亡相关蛋白的表达水平,评价糖尿病大血管焦亡损伤,及药物抗糖尿病大血管焦亡损伤的作用机制。结果表明,药物联合饮食诱导成功构建糖尿病模型。与对照组相比,模型组大鼠空腹血糖含量升高,血浆中总胆固醇(total cholesterol,TC)、甘油三酯(triglyceride,TG)、低密度脂蛋白胆固醇(low-density lipoprotein cholesterol,LDL-c)含量显著升高,高密度脂蛋白胆固醇(high-density lipoprotein cholesterol,HDL-c)含量显著降低,血管内膜增厚,血清与主动脉中炎症因子肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)、白细胞介素-1β(interleukin-1β,IL-1β)、白细胞介素-18(interleukin-18,IL-18)含量增加,炎症小体NLRP3表达增加,主动脉血管细胞中caspase-1、GSDMD活化蛋白增加。与模型组相比,黄精干预可降低血糖、血脂含量,抑制血管增厚,减少糖尿病大鼠血清及主动脉炎症因子TNF-α、IL-1β、IL-18表达,抑制炎症小体NLRP3表达以及焦亡相关蛋白caspase-1、GSDMD的活化。综上所述,黄精可通过抑制细胞焦亡,减轻血管局部炎症,延缓糖尿病大血管焦亡损伤进程。

This study aims to explore the influence of Polygonati Rhizoma on the pyroptosis in the rat model of diabetic macroangiopathy via the NOD-like receptor thermal protein domain associated protein 3(NLRP3)/cysteinyl aspartate specific proteinase-1(caspase-1)/gasdermin D(GSDMD)pathway.The rat model of diabetes was established by intraperitoneal injection of streptozotocin(STZ)combined with a high-fat,high-sugar diet.The blood glucose meter,fully automated biochemical analyzer,hematoxylin-eosin(HE)staining,enzyme-linked immunosorbent assay,immunofluorescence,immunohistochemistry,and Western blot were employed to measure blood glucose levels,lipid levels,vascular thickness,inflammatory cytokine levels,and expression levels of pyroptosis-related proteins.The mechanism of pharmacological interventions against the injury in the context of diabetes was thus explored.The results demonstrated the successful establishment of the model of diabetes.Compared with the control group,the model group showed elevated levels of fasting blood glucose,total cholesterol(TC),triglycerides(TG)and low-density lipoprotein cholesterol(LDL-c),lowered level of high-density lipoprotein cholesterol(HDL-c),thickened vascular intima,and elevated serum and aorta levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and interleukin-18(IL-18).Moreover,the model group showed increased NLRP3 inflammasomes and up-regulated levels of caspase-1 and GSDMD in aortic vascular cells.Polygonati Rhizoma intervention reduced blood glucose and lipid levels,inhibited vascular thickening,lowered the levels of TNF-α,IL-1β,IL-18 in the serum and aorta,attenuated NLRP3 inflammasome expression,and down-regulated the expression levels of caspase-1 and GSDMD,compared with the model group.In summary,Polygonati Rhizoma can slow down the progression of diabetic macroangiopathy by inhibiting pyroptosis and alleviating local vascular inflammation.