同型半胱氨酸硫内酯对缺氧神经母细胞瘤系细胞内质网应激途径相关因子表达的影响

Effect of homocysteine thiolactone on expression of endoplasmic reticulum stress pathway-related factors in hypoxic neuroblastoma cells

目的探讨同型半胱氨酸硫内酯(HTL)对缺氧神经母细胞瘤系细胞内质网应激途径相关因子表达的影响。方法选取人神经母细胞瘤株SHSY5Y细胞采用氯化钴作用24 h制作缺氧模型后,分为模型组、试验A组、试验B组、试验C组、试验D组,每组5例,后4组分别加入HTL 50、100、200和400μmol/L同时刺激。另取处于对数生长期的SHSY5Y细胞为对照组(n=5)。检测各组细胞活性,免疫组织化学染色观察模型组和试验D组、半胱氨酸天冬氨酸酶(Caspase)-12和Caspase-3及TNF-α蛋白表达,ELISA检测各组上述3种蛋白表达。结果与对照组比较,模型组细胞活力明显降低(0.922±0.039 vs 1.143±0.076,P=0.000)。试验A组、试验B组、试验C组、试验D组细胞活力明显低于模型组(P<0.01)。免疫组织化学染色显示,与模型组比较,试验D组TNF-α蛋白表达明显升高(0.34±0.04 vs 0.26±0.01,P=0.014)。模型组、试验A组~D组的Caspase-3、Caspase-12、TNF-α蛋白表达呈上升趋势。与对照组比较,试验C组Caspase-3和试验D组Caspase-3、Caspase-12、TNF-α蛋白表达明显升高(P<0.01)。结论HTL可能通过Caspase-12凋亡途径及TNF-α激活,抑制细胞生长,促进细胞炎性反应发生及凋亡,加重缺氧后损伤。

Objective To study the effect of homocysteine thiolactone(HTL)on expression of endoplasmic reticulum stress pathway-related factors in hypoxic neuroblastoma cells.Methods The human neuroblastoma SHSY5Y cells were divided into model group and test groups A-D which were treated with cobalt chloride,50,100,200 and 400μmol/L of HTL+cobalt chloride respectively with human neuroblastoma SHSY5Y cells in logarithmic growth stage served as a control group.The cell activity and the expressions of caspase-12,caspase-3,TNF-αin different groups were detected.Results The activity of human neuroblastoma SHSY5Y cells was significantly lower in model group than in control group and in test groups A-D than in model group(P<0.01).The expression level of TNF-αprotein was significantly higher in group D than in model group(0.34±0.04 vs 0.26±0.01,P=0.014).The expression levels of caspase-3,caspase-12 and TNF-αprotein were significantly higher in model group and test groups A-D and were significantly higher in test groups C and D than in control group(P<0.01).Conclusion HTL can inhibit the growth and inflammatory reaction respone of SHSY5Y cells and further aggravate their injury by regulating the caspase-12 apoptosis pathway and activating the TNF-α.