【摘要】：目的探討過表達(dá)骨形成蛋白和激活素的跨膜抑制劑(BMP and activin membrane-bound inhibitor,BAMBI)對谷氨酸誘導(dǎo)的大鼠脊髓背角神經(jīng)元和星形膠質(zhì)細(xì)胞凋亡的影響。方法原代培養(yǎng)新生Wistar大鼠脊髓背角神經(jīng)元和星形膠質(zhì)細(xì)胞,將細(xì)胞分為6組:正常對照組,谷氨酸組,空載組,BAMBI過表達(dá)組,si RNA對照組和BAMBI沉默組。Western blot檢測各組細(xì)胞中BAMBI的表達(dá),流式檢測細(xì)胞凋亡,Western blot檢測自噬標(biāo)志分子微管相關(guān)蛋白輕鏈3(microtubule associated protein light chain 3,LC3),Beclin1和p62的表達(dá),熒光顯微鏡檢測GFP-LC3的含量,最后Western blot檢測腫瘤壞死因子-α(TNF-α),白介素-1β(IL-1β),裂解后的半胱天冬酶(cleaved-Caspase3)的表達(dá)。結(jié)果與對照組相比,谷氨酸誘導(dǎo)的細(xì)胞凋亡增加,LC3-Ⅰ的表達(dá)降低,LC3-Ⅱ,Beclin1和p62的表達(dá)增加,GFP-LC3的含量增加,TNF-α、IL-1β和cleaved-Caspase3的表達(dá)增加(P0.05);與谷氨酸組相比,轉(zhuǎn)染pc DNABAMBI的細(xì)胞凋亡減少,LC3和Beclin1的表達(dá)顯著增加,LC3-Ⅰ和p62的表達(dá)降低,細(xì)胞內(nèi)可見大量的GFP-LC3堆積,TNF-α、IL-1β和cleaved-Caspase3的表達(dá)顯著下降(P0.05)。BAMBI沉默組的結(jié)果與過表達(dá)組相反。結(jié)論過表達(dá)BAMBI可增強自噬,抑制谷氨酸誘導(dǎo)的大鼠脊髓背角神經(jīng)元和星形膠質(zhì)細(xì)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- 偽 (TNF- 偽), 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，

本文編號：2356104