甲基苯丙胺对大鼠小胶质细胞的损伤及炎性相关基因表达的影响

发布时间：2018-05-06 05:26

本文选题：甲基苯丙胺 + 小胶质细胞　；参考：《南京医科大学学报(自然科学版)》2017年09期

【摘要】：目的:观察甲基苯丙胺(methamphetamine,Meth)对原代小胶质细胞的损伤及炎性相关基因表达的影响。方法:原代培养SD胎鼠小胶质细胞,利用MTT和原位末端转移酶标记技术(TUNEL)分别检测Meth引起小胶质细胞活力和凋亡的变化。采用实时荧光定量聚合酶链反应法(real-time PCR)观察Meth对小胶质细胞炎性相关因子mRNA表达的影响。ELISA及试剂盒法检测Meth作用后小胶质细胞白介素(interleukin,IL)-6、肿瘤坏死因子(tumor necrosis factor,TNF)-α、诱导型一氧化氮合酶(inducible nitric oxide synthase,i NOS)的分泌改变。结果:MTT实验显示,Meth降低小胶质细胞活力,呈浓度依赖性,浓度为200μmol/L时与对照组比较,差异有统计学意义(P0.01)。TUNEL实验结果显示200μmol/L Meth可引起细胞凋亡,与对照组比较,差异有统计学意义(P0.05)。q-PCR结果显示Meth作用于小胶质细胞24 h可降低IL-24、一氧化氮合酶3(nitric oxide synthase 3,NOS3)表达水平,上调Peli3、Sigma受体1(Sigma receptor 1,Sig1-R)、IL-1β、IL-6、Toll样受体4(Toll like receptor 4,TLR4)的表达水平,差异有统计学意义(P0.05)。此外,蛋白水平亦发现,Meth可促进IL-6和TNF-α的分泌。结论:Meth可降低小胶质细胞活力,诱导小胶质细胞凋亡,并引起IL-1β、IL-1R、IL-6、TLR4等炎性因子mRNA表达变化,促进IL-6和TNF-α的分泌,进而可能损伤中枢神经系统,产生神经毒性。
[Abstract]:Aim: to investigate the effect of methamphetamine methamphetamine (methamphetamine) on primary microglia injury and inflammatory related gene expression. Methods: primary cultured fetal SD rat microglia cells were cultured. The changes of microglial activity and apoptosis induced by Meth were detected by MTT and in situ terminal transferase labeling technique (Tunel). Real-time PCR was used to observe the effect of Meth on the expression of mRNA of microglial inflammatory related factors. Elisa and kit were used to detect interleukin-6, tumor necrosis factor- TNF- 伪 and tumor necrosis factor (TNF- 伪) in microglial cells after Meth treatment. The secretory changes of inducible nitric oxide synthase I NOS. Results the cell viability was decreased in a concentration-dependent manner, and the difference was significant when the concentration was 200 渭 mol/L. The results of Tunel showed that 200 渭 mol/L Meth could induce cell apoptosis, compared with the control group. The results showed that Meth could decrease the expression of IL-24, nitric oxide synthase (3(nitric oxide synthase 3) and up-regulate the expression of 1(Sigma receptor 1 Sigma receptor 4(Toll like receptor 4 TLR4 in microglia for 24 h, and the difference was statistically significant (P 0 05). In addition, the protein level also found that Meth can promote the secretion of IL-6 and TNF- 伪. ConclusionMeth can decrease the activity of microglia, induce apoptosis of microglia, and induce the expression of mRNA, such as IL-1 尾 -IL-1RL-6IL-6TLR4, and promote the secretion of IL-6 and TNF- 伪, which may damage the central nervous system and produce neurotoxicity.
【作者单位】：南京医科大学公共卫生学院卫生毒理系;南京医科大学第一附属医院急诊医学中心;
【基金】：国家自然科学基金(81673213,81202230) 江苏省自然科学基金(BK20151557) 江苏高校优势学科建设工程资助项目
【分类号】：R749.64

【相似文献】