普鲁泊福对TNF-α诱导的小鼠脊髓神经元凋亡及Bax表达的影响

In vitro effects of propofol on apoptosis and Bax expression induced by TNF-α in mouse spinal cord neurons

目的:研究普鲁泊福(propofol)对肿瘤坏死因子α(TNF-α)诱导的小鼠脊髓神经元凋亡及Bax表达的影响。方法:脊髓神经元取自小鼠胎鼠,置于含B27的神经细胞培养基中培养,在培养第7天随机分为6组:对照组,50μmol/L普鲁泊福组,TNF-α组,25μmol/L普鲁泊福+TNF-α组,50μmol/L普鲁泊福+TNF-α组,100μmol/L普鲁泊福+TNF-α组。在相应组中加入不同终浓度的普鲁泊福,孵育30 min,再加入TNF-α至终末浓度为2 000 U/ml,培养24 h后,采用碘化丙锭(PI)/Hoechst33342双染法检测细胞凋亡,于荧光显微镜下观察凋亡细胞。采用免疫细胞化学的方法检测Bax的表达。结果:与对照组相比,TNF-α组细胞凋亡百分率明显升高[(21.8±1.1)%vs(2.8±0.8)%,P<0.01],Bax表达明显增加[(0.251±0.016)vs(0.141±0.015),P<0.01];而经不同浓度普鲁泊福(25,50,100μmol/L)处理后再加入TNF-α的各组与TNF-α组相比,细胞凋亡百分率下降[(16.2±1.2)%、(15.3±0.6)%(、12.2±0.8)%vs(21.8±1.1)%,P<0.05、P<0.05、P<0.01],Bax的表达降低[(0.198±0.011)(、0.188±0.012)、(0.167±0.014)vs(0.251±0.016),P均<0.05],且呈现一定的剂量效应关系。结论:临床浓度的普鲁泊福可能通过调节Bax的表达进而抑制由TNF-α诱导的脊髓神经元凋亡。

Objective：To study the in vitro effects of propofol on apoptosis and Bax expression induced by TNF-α in mouse spinal cord neurons. Methods： Spinal cord neurons were isolated from fetal mice and cultured in neurobasal medium with B27 supplement. On the 7th day of culture, neurons were randomly divided into 6 groups： control group, propofol（50μmol/L） group, TNF-α group, propofol（25 μmol/L） with TNF-α group, propofol（50μmol/L） with TNF-α group and propofol（100μmol/L） with TNF-α group. Propofol with different concentrations was incubated with cultured cells for 30 min, then TNF-α was added with the final concentration of 2 000 U/ml for another 24 h incubation. Apoptosis was detected by PI/Hoechst33342 double staining technique and fluorescence microscopy. Bax expression was determined by immunocytochemical technique. Resuits： The apoptosis rate and expression of Bax in TNF-α group were both increased compared with those in control group （[21.8± 1.1]% vs [2.8±0.8]% ,P〈0.01; [0. 251±0. 016] vs [0. 141±0. 015] ,P〈0.01）. The apoptosis rates and expression of Bax in 3 propofol groups（25,50,100μmol/L） were （[16. 2 ± 1. 2]%, [15. 3± 0. 6]% and [12. 2 ± 0. 8]%） and （[0. 198±0. 011] ,[0. 188±0. 012] and [0. 167±0. 014]）,respectively,and they were all significantly decreased compared with those in TNF-α group （[21.8± 1.1] % and [0. 251± 0. 0163], all P〈0.05）. Moreover, a dose-dependent manner was observed in the above changes. Conclusion： Propofol within clinical concentration ranges may inhibit the apoptosis of spinal cord neurons induced by TNF-α through modulating the expression of Bax.