大黄糖络丸调控NLRP3/caspase-1/GSDMD信号通路改善糖尿病大鼠大血管炎症损伤

DaHuangTangLuoWan regulates NLRP3/caspase-1/GSDMD signal pathway to improve macrovascular inflammatory injury in diabetic rats

关键词:  糖尿病大血管病变  大黄糖络丸  炎症损伤  NLRP3/Caspase-1/GSDMD通路

Key words:  diabetic macroangiopathy  DaHuangTangLuoWan  inflammatory injury  NLRP3/Caspase-1/GSDMD pathway

摘要:

摘要：目的 明确大黄糖络丸对糖尿病大鼠大血管损伤的干预作用，并基于NLRP3/Caspase-1/GSDMD通路介导炎症损伤探讨其具体机制。方法 选取40只SPF级9周龄ZDF（纯合子fa/fa）大鼠进行实验，适应性喂养1周后采用Purina#5008饲料诱导3周复制模型，成模后随机分为模型对照组、二甲双胍组（0.09g/Kg）和大黄糖络丸高、中、低剂量组（2.16、1.08、0.54g/kg），另选8只同龄ZDF（杂合子fa/+）大鼠作为空白对照组，干预12周后测定大鼠体质量和空腹血糖。采用全自动生化分析仪检测血浆中TC、TG、LDL-c、HDL-c的含量；采用HE染色法观察大鼠血管组织病理形态变化；采用ELISA技术检测血清中IL-1β和IL-18的含量；采用WB与qPCR技术检测血管组织中NLRP3、ASC、Caspase-1、GSDMD 蛋白和mRNA表达水平。结果 与空白对照组相比，模型对照组大鼠体质量和空腹血糖含量显著升高（P<0.05），TC、TG、LDL-c含量显著升高（P<0.05），HDL-c含量显著降低（P<0.05），血管内细胞排列不规则且出现细胞空泡，血管内膜增厚，IL-1β、IL-18含量显著增加（P<0.05），NLRP3、Caspase-1、GSDMD 蛋白和mRNA表达增加（P<0.05）。与模型对照组相比，各剂量组大鼠体质量和空腹血糖均显著降低（P<0.05），TC、TG、LDL-c含量显著升高（P<0.05），HDL-c含量显著降低（P<0.05），不同程度改善了血管病理损伤，血清中炎症因子（IL-1β、IL-18）含量显著降低（P<0.05），NLRP3、Caspase-1、GSDMD 的蛋白与mRNA表达显著降低（P<0.05）。与大黄糖络丸低剂量组相比，大黄糖络丸高剂量组有更加显著的改善作用（P<0.05）。结论 大黄糖络丸能够显著改善糖尿病大鼠血管损伤，并呈显著的剂量依赖，其具体机制与大黄糖络丸抑制NLRP3/Caspase-1/GSDMD通路改善血管炎症性损伤有关。

Abstract:

ABSTRACT: OBJECTIVE To clarify the interventional effects of DaHuangTangLuoWan (DHTLW) on macrovascular injury in diabetic rats and to explore the specific mechanisms based on the NLRP3/Caspase-1/GSDMD pathway mediating inflammatory injury. METHODS Forty SPF-grade 9-week-old ZDF (purebred fa/fa) rats were selected for the experiment, and the replica model was induced by Purina#5008 chow for 3 weeks after 1 week of acclimatization feeding, and then randomly divided into a model control group, a metformin group (0.09g/Kg), and a high-, medium-, and low-dose group of DHTLW (2.16, 1.08, and 0.54 g/kg) after the model was formed. Another 8 ZDF (heterozygous fa/+) rats of the same age were selected as blank control group, and the body mass and fasting blood glucose of the rats were measured after 12 weeks of intervention. The plasma levels of TC, TG, LDL-c and HDL-c were detected using an automatic biochemical analyzer; the pathomorphological changes of rat vascular tissues were observed using HE staining; the serum levels of IL-1β and IL-18 were detected using ELISA; and the expression levels of NLRP3, ASC, Caspase-1 and GSDMD proteins and mRNAs were detected by using WB and qPCR techniques. protein and mRNA expression levels. RESULTS Compared with the blank control group, rats in the model control group showed significantly higher body mass and fasting glucose content (P<0.05), significantly higher TC, TG, LDL-c content (P<0.05), significantly lower HDL-c content (P<0.05), irregular cell arrangement and cell vacuoles in the vasculature, thickening of vascular endothelium, and significant increase in the content of IL-1β, IL-18 ( P<0.05), increased expression of NLRP3, Caspase-1, GSDMD protein and mRNA (P<0.05). Compared with the model control group, body mass and fasting blood glucose of rats in each dose group were significantly reduced (P<0.05), TC, TG, LDL-c content was significantly increased (P<0.05), HDL-c content was significantly reduced (P<0.05), vascular pathological injury was improved to different degrees, and the content of serum inflammatory factors (IL-1β, IL-18) was significantly reduced (P<0.05), protein and mRNA expression of NLRP3, Caspase-1, GSDMD were significantly reduced (P<0.05). Compared with the low-dose group, the high-dose group of DHTLW showed more significant improvement (P<0.05). CONCLUSION DHTLW can significantly improve vascular injury in diabetic rats in a dose-dependent manner, and the specific mechanism is related to the inhibition of NLRP3/Caspase-1/GSDMD pathway by DHTLW to improve vascular inflammatory injury.