NLRP3在糖尿病肾病病理条件下的表达变化研究.docVIP

  NLRP3 在糖尿病肾病病理条件下的表达变 化研究# 都鹏超，王宇芃，范宝霞，韩慧蓉，宋振宇，易凡** 5 10 15 20 25 30 35 40 （山东大学医学院药理学研究所，济南 250012） 摘要：目的：通过对糖尿病肾病（diabetic nephropathy, DN）病人临床穿刺样本和 DN 小鼠 模型中肾脏组织以及肾脏细胞在模拟糖尿病肾病条件刺激下的体外研究，探讨 NLRP3 炎症 体在糖尿病肾病病理条件下的表达变化。方法：通过对 DN 病人穿刺样本进行免疫组化和 real-time RT-PCR 检测，观察 NLRP3 在 DN 病人穿刺样本中的表达变化；通过高脂喂养（HFD） /STZ 尾静脉注射联合建立 DN 小鼠模型，并通过 Western Blot 和半定量 PCR 方法观察 NLRP3 在 DN 小鼠模型中的表达变化；在体外实验中进一步探讨足细胞和系膜细胞在高糖、TNF- α、TGF-β条件下 NLRP3 蛋白的表达。结果：DN 病人小鼠模型中肾脏组织中的 NLRP3 表 达水平显著升高；足细胞在高糖、TNF-α或 TGF-β刺激下 NLRP3 表达明显升高且具有统 计学意义，而系膜细胞在高糖刺激下 NLRP3 蛋白表达升高，在 TNF-α、TGF-β刺激下 NLRP3 蛋白表达没有显著的变化。结论：NLRP3 炎症体的表达上调在 DN 的发病机制中可 能发挥重要作用，对肾脏固有细胞的体外实验表明 NLRP3 的表达变化具有细胞特异性，深 入研究 NLRP3 炎症体的作用机制将为糖尿病肾病的治疗提供新的靶点和思路。 关键词：药理学；NLRP3 炎症体；糖尿病肾病；固有免疫；足细胞 中图分类号：R966 The expression patterns of NLRP3 in diabetic nephropathy DU Pengchao, WANG Yupeng, FAN Baoxia, HAN Huirong, SONG Zhenyu, YI Fan (Shandong University, School of Medicine 250012) Abstract: Aim: The present study was designed to detect the expression of NLRP3 in diabetic nephropathy (DN) including DN clinical renal biopsies and the kidneys from DN mice and renal cells under the stimulation similar as diabetic environment. Methods: Immunohistochemistry and real-time RT-PCR were used to examine the expression patterns of NLRP3 in the DN clinical renal biopsies compared with normal controls. DN mice models were induced by high-fat-diet (HFD) and streptozotocin (STZ) injection. Western blot, semiquantitative RT-PCR were used to examine the expression changes of NLRP3 in the DN mice and the expression of NLRP3 in podocytes and mesangial cells treated with high glucose, TNF-α, TGF-β in vitro. Results: There was a significant increase in NLRP3 level in the kidneys from DN patients and DN mice; NLRP3 expression was significantly high in podocytes under the stimulation of high glucose, TNF-α or TGF-β. However, NLRP3 expression increased in high glucose-treated mesangial cells rather than TNF-α or TGF-β treatment. Conclusion: NLRP3 may play an important role