首页|钩藤碱对甲基苯丙胺依赖SH-SY5Y细胞模型及miR-375-3p/Elavl4表达的影响

钩藤碱对甲基苯丙胺依赖SH-SY5Y细胞模型及miR-375-3p/Elavl4表达的影响

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  • 国家科技期刊平台
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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.
Effects of rhynchophylline on methamphetamine-dependent SH-SY5Y cells model and expression of miR-375-3p/Elavl4
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.

methamphetaminedrug dependencerhynchophyllinemicroRNA-375-3p/embryonic lethal abnormal vision drosophila-like 4

李汉成、曾茗、陈依玲、陈玲红、莫志贤

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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

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

国家自然科学基金广东省重点领域研发计划广东省科技厅农村科技特派员课题广东省教育厅青年创新人才基金肇庆市科技指导类基金肇庆学院博士科研启动基金肇庆学院科研项目肇庆学院大学生创新创业训练计划肇庆学院大学生创新创业训练计划

818730302021B0707010008KTP202001762023kQNCX0902023040316003220003QN202224S202310580075S202310580070

2024

10.13699/j.cnki.1001-6821.2024.01.010

中国临床药理学杂志
中国药学会

中国临床药理学杂志

CSTPCD北大核心
影响因子：1.91
ISSN：1001-6821
年,卷(期)：2024.40(1)
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