目的 探讨钩藤碱对甲基苯丙胺依赖人神经母细胞瘤细胞(SH-SY5Y细胞)模型及微小RNA-375-3p(miR-375-3p)/胚胎致死异常视觉-样4蛋白(Elav14)表达的影响.方法 用最大安全剂量诱导法建立甲基苯丙胺依赖细胞模型.将细胞分成正常组(完全培养液)、对照组(完全培养液+400 μmol·L-1钩藤碱孵育48 h)、模型组(完全培养液+100 μmol·L-1甲基苯丙胺孵育48 h)、实验组(完全培养液+400 μmol·L-1的钩藤碱孵育15 min后,再加入100μmol·L-1甲基苯丙胺孵育48 h).以酶联免疫吸附法检测细胞环腺苷酸(cAMP)、5-羟色胺(5-HT)水平;以实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测细胞miR-375-3p表达情况;以蛋白质印迹法检测Elavl4蛋白表达情况.结果 正常组、对照组、模型组和实验组cAMP表达水平分别为(6.33±0.93)、(6.57±1.12)、(10.89±1.03)和(7.81±1.32)pmol·mg-1;5-HT分别为(682.46±17.32)、(690.31±15.09)、(510.11±27.67)和(649.99±21.42)pg·mL-1;miR-375-3p 表达水平分别为 1.00±0.13、1.13±0.24、3.48±0.18 和 1.58±0.19;Elavl4 表达水平分别为 1.00±0.05、0.89±0.10、0.50±0.09和0.90±0.11.模型组上述指标与正常组比较,差异均有统计学意义(P＜0.01,P＜0.05);实验组上述指标与模型组比较,差异均有统计学意义(P＜0.05,P＜0.01).结论 本研究建立甲基苯丙胺依赖细胞模型,同时发现钩藤碱可能调控miR-375-3p/Elavl4表达拮抗甲基苯丙胺成瘾.
Objective To investigate the effects of rhynchophylline on methamphetamine-dependent SH-SY5Y cells model and microRNA-375-3p(miR-375-3p)/embryonic lethal abnormal vision drosophila-like 4(Elavl4)expression.Methods A methamphetamine-dependent cell model by maximum safe dose induction was established.The cells were divided into normal group(complete culture medium),control group(complete culture medium+400 μmol·L-1 rhynchophylline incubated for 48 h),model group(complete culture medium+100 μmnol·L-1 methamphetamine incubated for 48 h)and experimental group(complete culture medium+400 μmol·L-1 rhynchophylline incubated for 15 min,then 100 μmol·L-1 methamphetamine incubated for 48 h).The cyclic adenosine monophosphate(cAMP)and 5-hydroxytryptamine(5-HT)expression were detected by enzyme-linked immunosorbent assay(ELISA),miR-375-3p expression was detected by quantitative real time polymerase chain reaction(qRT-PCR),and Elavl4 expression was detected by Western blot.Results The cAMP expression levels of normal group,control group,model group and experimental group were(6.33±0.93),(6.57±1.12),(10.89±1.03)and(7.81±1.32)pmol·mg-1;5-HT were(682.46±17.32),(690.31±15.09),(510.11±27.67)and(649.99±21.42)pg·mL-1;miR-375-3p expression were 1.00±0.13,1.13±0.24,3.48±0.18 and 1.58±0.19;Elavl4 expression were 1.00±0.05,0.89±0.10,0.50±0.09 and 0.90±0.11,respectively.The differences between above indicators in model group and normal group were statistically significant(P＜0.01,P＜0.05);the differences between above indicators in experimental group and model group were statistically significant(P＜0.01,P＜0.05).Conclusion This study preliminarily established a methamphetamine-dependent cell model,and also found that rhynchophylline may regulate miR-375-3p/Elavl4 expression to antagonize methamphetamine addiction.
李汉成;曾茗;陈依玲;陈玲红;莫志贤
肇庆学院食品与制药工程学院,广东肇庆 526061南方医科大学中医药学院,广东广州 510515
中医学
甲基苯丙胺;药物依赖;钩藤碱;微小RNA-375-3p/胚胎致死异常视觉-样4蛋白
methamphetamine;drug dependence;rhynchophylline;microRNA-375-3p/embryonic lethal abnormal vision drosophila-like 4
《中国临床药理学杂志》 2024 (001)
47-51 / 5
国家自然科学基金资助项目(81873030);广东省重点领域研发计划基金资助项目(2021B0707010008);广东省科技厅农村科技特派员课题资助项目(KTP20200176);广东省教育厅青年创新人才基金资助项目(2023kQNCX090);肇庆市科技指导类基金资助项目(2023040316003);肇庆学院博士科研启动基金资助项目(220003);肇庆学院科研基金资助项目(QN202224);肇庆学院大学生创新创业训练计划基金资助项目(S202310580075,S202310580070)
10.13699/j.cnki.1001-6821.2024.01.010
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