基于NLRP3/GSDMD信号通路探索大黄素对糖尿病肾病大鼠肾组织的修复

Elucidating the Renoprotective Effects of Emodin in Diabetic Kidney Disease Rats via the NLRP3/GSDMD Signaling Pathway

关键词:  糖尿病肾病  大黄素  焦亡  NLRP3/GSDMD信号通路

Key words:  diabetic kidney disease  emodin  pyroptosis  NLRP3/GSDMD signaling pathway

摘要:

摘要：目的：基于NLRP3/GSDMD信号通路探索大黄素（Emodin，EM）对链脲佐菌素（Streptozotocin,STZ)诱导大鼠糖尿病肾病（Diabetic Kidney Disease，DKD)模型焦亡的影响。方法：设置对照组，单侧肾切除联合单次腹腔注射STZ诱导动物模型，分为糖尿病肾病模型组、大黄素干预组(40 mg·kg-1、80 mg·kg-1）及NLRP3抑制剂组（MCC950,10 mg·kg-1）。检测各组治疗前后肾功能指标并动态记录体重及血糖值变化；过碘酸-希夫染色（Periodic Acid-Schiff Stain，PAS）和马松三色染色（Masson）观察动物肾脏受损及修复情况；RT-qPCR检测IL-18、IL-1β、HO-1及NPHS2基因的表达水平变化；Western Blot检测Nephrin、NPHS2、CD2AP、NLRP3、GSDMD及caspase-1蛋白表达量变化。结果：经大黄素干预的DKD大鼠各项肾功能指标均得到改善；与模型组相比，80 mg·kg-1的大黄素干预显著降低了血清肌酐（Serum Creatinine,Scr）、血尿素氮（Blood Urea Nitrogen,BUN）等各项肾功能指标（p均＜0.05）；Nephrin、NPHS2、CD2AP等蛋白含量显著上调（p均＜0.05），NLRP3、GSDMD、caspase-1蛋白表达含量均显著下调（p均＜0.05）；IL-18、IL-1β等炎症因子含量出现下降趋势（p均＜0.05），并且NPHS2及HO-1含量也出现不同程度的上升（p均＜0.05）。结论：大黄素通过调节NLRP3/GSDMD焦亡信号通路，改善了大鼠的肾功能及病理损伤。

Abstract:

ABSTRACT: OBJECTIVE To investigate the effect of emodin (EM) on pyroptosis in a rat model of diabetic kidney disease (DKD) induced by streptozotocin (STZ), based on the NLRP3/GSDMD signaling pathway. METHODS Rats were divided into a control group and a model group induced by unilateral nephrectomy combined with a single intraperitoneal injection of STZ. The model animals were further assigned to a DKD model group, emodin intervention groups (40 mg·kg-1, 80 mg·kg-1), and an NLRP3 inhibitor group (MCC950, 10 mg·kg-1). Renal function indicators were measured before and after treatment, and changes in body weight and blood glucose levels were dynamically recorded. Kidney injury and repair were observed using Periodic Acid-Schiff Stain and Masson staining. The mRNA expression levels of IL-18, IL-1β,HO-1 and NPHS2 were detected by RT-qPCR.Protein expression levels of Nephrin、NPHS2、CD2AP、NLRP3、GSDMD and caspase-1 were measured by Western blot.RESULTS Emodin intervention improved various renal function indicators in DKD rats. Compared with the model group, 80 mg·kg-1 emodin significantly reduced serum creatinine(Scr) and blood urea nitrogen(BUN) levels (all p < 0.05). Protein levels of Nephrin, NPHS2, and CD2AP were significantly up-regulated (all p < 0.05), while the expression of NLRP3, GSDMD, and caspase-1 was significantly down-regulated (all p < 0.05). Inflammatory factors such as IL-18 and IL-1β showed a decreasing trend (all p < 0.05), and the expression of NPHS2 and HO-1 increased to varying degrees (all p < 0.05).CONCLUSION Emodin improves renal function and pathological damage in DKD rats by regulating the NLRP3/GSDMD-mediated pyroptosis signaling pathway.