二乙酰吗啡对BV-2细胞损伤作用机制研究  

Damage mechanism of diacetylmorphine on BV-2 cells

作　　者：谢明仁 祁珊 俞蕾 袁霞 杜靖 俞发荣 Xie Mingren;Qi Shan;Yu Lei;Yuan Xia;Du Jing;Yu Farong(Public Security Branch,Gansu University of Political Science and Law,Gansu Lanzhou,730070,China)

机构地区：[1]甘肃政法大学公安分院,甘肃兰州730070

出　　处：《中国法医学杂志》2024年第3期304-307,314,共5页Chinese Journal of Forensic Medicine

基　　金：甘肃省自然科学基金项目(21JR7RA571);甘肃政法大学重点项目(GZF2021XZD06);甘肃政法大学校级科研创新团队项目;兰州市科技计划项目(2022-2-68)。

摘　　要：目的 探索二乙酰吗啡(DAM)对神经胶质细胞(BV-2)损伤作用机制。方法 选用具有分裂增殖能力的神经胶质细胞(BV-2)为实验对象,分别给予30、60、120 mg/L DAM,孵育4、8、16、32和48 h,用台盼蓝、噻唑蓝(MTT)法检测DAM对BV-2细胞损伤程度和增殖抑制率,用流式细胞仪检测DAM对BV-2细胞周期和细胞凋亡的影响,用ELISA法检测BV-2细胞BDNF、HSP-70、Bax、caspase-9、c-Fos、TrkB蛋白水平。结果 台盼蓝、MTT检测显示,死亡细胞依DAM浓度增加和作用时间的延长显著增多,BV-2细胞增殖抑制率比对照组显著升高。流式细胞仪检测显示,DAM组G0/G1期细胞数比对照组明显增加,G2期细胞数显著减少,凋亡率增加。ELISA法检测显示,DAM组Bax、caspase-9、c-Fos蛋白水平比对照组显著升高,HSP-70、BDNF、TrkB蛋白水平比对照组明显降低。结论 DAM对BV-2细胞损伤作用机制与上调损伤蛋白水平、下调保护蛋白水平、损伤细胞膜结构、抑制细胞分裂、导致细胞凋亡、死亡有关。Objective To study the damage mechanism of diacetylmorphine(DAM) on BV-2cells.Methods BV-2cells with the ability to divide and proliferate were selected as experimental objects,BV-2 cells were treated with 30,60and 120 mg/L DAM,respectively.The cells were cu?tured for 4,8,16,32 and 48 h.The damage degree and proliferation inhibition rate of BV-2 cells were detected by trypan blue and thiazole blue(MTT) method.The effects of DAM on BV-2cell cycle and apoptosis were detected with flow cytometry.The c-Fos,Bax,caspase-9,BDNF,HSP-70 and TrkB protein level in BV-2 cells were detected by ELISA.Results Trypan blue and MTT detection showed that the death cells increased significantly with the increase of DAM concentration and prolonged action time and the inhibition rate of BV-2 cells was significantly higher than that of the control group.Cell cycle showed that the number of G0/G1 phase cells in DAM group was significantly increased,the number of G2 phase cells was significantly decreased,and the apoptosis rate was increased.ELISA showed that the protein levels of caspase-9,c-Fos and Bax in DAM groups were significantly higher than those in the control group.On the contrary,BDNF,HSP-70 and TrkB protein levels were significantly lower than those in the control group.Conclusion The damage mechanism of DAM on BV-2 cells is related to up-regulating damage protein level,downregulating protection protein level,damaging cell membrane structure,inhibiting cell division,leading to cell apoptosis and death.

关 键 词：法医病理学 损伤作用 二乙酰吗啡 BV-2细胞 凋亡 

分 类 号：D919.1[医药卫生—法医学]