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作 者:王姝瑶[1] 孟天娇 姚墨 刘佳[4] 叶芸 姚尚争 刘影 孙文萍 Wang Shuyao;Meng Tianjiao;Yao Mo;Liu Jia;Ye Yun;Yao Shangzheng;Liu Ying;Sun Wenping(Department of Neurology,Tongliao Hospital,Tongliao 028000,Inner Mongolia Autonomous Region,China)
机构地区:[1]内蒙古通辽市医院神经内科,028000 [2]内蒙古通辽市医院骨科,028000 [3]诸暨市人民医院神经内科 [4]首都医科大学宣武医院神经内科 [5]北华大学附属医院神经内科 [6]上海市松江区中心医院全科医学科
出 处:《中华老年心脑血管病杂志》2020年第2期190-192,共3页Chinese Journal of Geriatric Heart,Brain and Vessel Diseases
基 金:吉林省科技发展计划(20180101144JC);吉林省教育厅“十三五”科学技术项目(JJKH20170063KJ);吉林省卫生技术创新项目(2017J079)
摘 要: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.
关 键 词:神经母细胞瘤 内质网应激 肿瘤坏死因子Α 半胱氨酸天冬氨酸蛋白酶12
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