PURα介导糖尿病神经病变大鼠发病神经元中p27<sup>kip-1</sup>和TNF-α的表达及其意义-东南大学学报医学版

PURα介导糖尿病神经病变大鼠发病神经元中p27^kip-1和TNF-α的表达及其意义

单位：华中科技大学同济医学院附属武汉中心医院内分泌科, 湖北武汉 430000

关键词：糖尿病神经病变人转录因子活性蛋白α p27^kip-1 肿瘤坏死因子α 大鼠转染

分类号：R587.2;R-33

出版年·卷·期（页码）：2019·38·第三期（427-432）

摘要：

目的：探讨人转录因子活性蛋白α（PURα）介导糖尿病神经病变（DN）神经元中抑制蛋白p27^kip-1和肿瘤坏死因子α（TNF-α）的表达及其意义。方法：采用腹膜腔注射链脲佐菌素法制备DN大鼠作为DN组，以健康大鼠为对照组，每组20只。比较1周内两组大鼠病理性疼痛和热痛觉过敏情况，分析两组大鼠背根节（DRG）神经元PURα、p27^kip-1和TNF-α的表达。取鼠源海马神经元细胞株HT-22，转染为4组：PURα过表达组（转染PURα表达质粒）、mock组（不转染任何表达载体）、空白对照组（转染空表达载体）及PURα沉默组（转染PURαRNAi表达质粒），分析细胞株PURα、p27^kip-1和TNF-α的表达。结果：与对照组比较，DN组大鼠出现明显的触觉异常性疼痛和热痛觉过敏（P<0.05），DN组大鼠DRG神经元PURα、p27^kip-1及TNF-α的表达上调（P<0.05），PURα与p27^kip-1及TNF-α的表达呈显著正相关（P<0.05）。神经元细胞株中上调PURα的表达能显著提高p27^kip-1及TNF-α的表达水平（P<0.05），抑制细胞内源PURα表达，p27^kip-1及TNFα的表达水平则显著降低（P<0.05）。结论：PURα可介导p27^kip-1和TNF-α蛋白表达，可能参与了DN的发病过程。

Objective:To explore the purine-rich element binding protein alpha(PURα)-mediated expression of p27^kip-1and tumor necrosis factor alpha(TNF-α) in the pathogenesis of diabetic neuropathy(DN). Methods:DN rats were prepared by intraperitoneal injection of streptozotocin as the DN group, and healthy rats were used as the control group with 20 rats in each group. The pathological pain and thermal hyperalgesia were compared between the two groups within one week. The expression levels of PURα, p27^kip-1 and TNF-α in dorsal root ganglion(DRG) neurons in the two groups were analyzed by Western blot. The mouse hippocampal neuron cell line HT-22 was transfected into 4 groups,i.e., the PURα overexpression group (transfected with PURα expression plasmid), the mock group(without transfection of any expression vector),the blank control group (transfected empty expression vector) and the PURα silencing group (transfected with PURα RNAi expression plasmid), and the expression of PURα, p27^kip-1 and TNF-α in the four cell lines groups were determined. Results:Compared with the control group, the tactile allodynia and thermal hyperalgesia were increased,and the expression levels of PURα, p27^kip-1 and TNF-α was up-regulated in DRG neurons of the DN group(P<0.05). PURα was significantly and positively correlated with the expression of p27^kip-1 and TNF-α(P<0.05). Up-regulation of PURα expression in neuronal cell lines significantly increased the expression of p27^kip-1 and TNF-α(P<0.05).As the expression of endogenous PURα was inhibited, the expression levels of p27^kip-1 and TNF-α were significantly decreased(P<0.05). Conclusion:PURα can mediate the expression of p27^kip-1 and TNF-α protein, which may be involved in the pathogenesis of DN.

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