骨形成蛋白和激活素的跨膜抑制剂调节自噬抑制谷氨酸诱导的大鼠脊髓背角神经组织细胞凋亡

发布时间：2018-11-25 12:27

【摘要】：目的探讨过表达骨形成蛋白和激活素的跨膜抑制剂(BMP and activin membrane-bound inhibitor,BAMBI)对谷氨酸诱导的大鼠脊髓背角神经元和星形胶质细胞凋亡的影响。方法原代培养新生Wistar大鼠脊髓背角神经元和星形胶质细胞,将细胞分为6组:正常对照组,谷氨酸组,空载组,BAMBI过表达组,si RNA对照组和BAMBI沉默组。Western blot检测各组细胞中BAMBI的表达,流式检测细胞凋亡,Western blot检测自噬标志分子微管相关蛋白轻链3(microtubule associated protein light chain 3,LC3),Beclin1和p62的表达,荧光显微镜检测GFP-LC3的含量,最后Western blot检测肿瘤坏死因子-α(TNF-α),白介素-1β(IL-1β),裂解后的半胱天冬酶(cleaved-Caspase3)的表达。结果与对照组相比,谷氨酸诱导的细胞凋亡增加,LC3-Ⅰ的表达降低,LC3-Ⅱ,Beclin1和p62的表达增加,GFP-LC3的含量增加,TNF-α、IL-1β和cleaved-Caspase3的表达增加(P0.05);与谷氨酸组相比,转染pc DNABAMBI的细胞凋亡减少,LC3和Beclin1的表达显著增加,LC3-Ⅰ和p62的表达降低,细胞内可见大量的GFP-LC3堆积,TNF-α、IL-1β和cleaved-Caspase3的表达显著下降(P0.05)。BAMBI沉默组的结果与过表达组相反。结论过表达BAMBI可增强自噬,抑制谷氨酸诱导的大鼠脊髓背角神经元和星形胶质细胞凋亡。
[Abstract]:Aim to investigate the effects of transmembrane inhibitor (BMP and activin membrane-bound inhibitor,BAMBI (a transmembrane inhibitor of bone morphogenetic protein and activin) on glutamate induced apoptosis of spinal dorsal horn neurons and astrocytes in rats. Methods Spinal dorsal horn neurons and astrocytes of newborn Wistar rats were cultured and divided into 6 groups: normal control group, glutamate group, no-load group, BAMBI overexpression group. The expression of BAMBI, Beclin1 and p62 were detected by. Western blot in si RNA control group and BAMBI silencing group, apoptosis, Western blot was detected by flow cytometry, and the expression of microtubule-associated protein light chain 3 (microtubule associated protein light chain 3mLC3), Beclin1 and p62 were detected by flow cytometry. The content of GFP-LC3 was detected by fluorescence microscope, and the expression of tumor necrosis factor- 伪 (TNF- 伪), interleukin-1 尾 (IL-1 尾) and cysteinase (cleaved-Caspase3) were detected by Western blot. Results compared with control group, glutamate induced apoptosis increased, LC3- 鈪，

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