海洛因通过下调DNMT3a抑制印记基因H19启动子甲基化促进雄鼠的子代成瘾与复吸的机制  

Mechanism of heroin promoting addiction and relapse in progeny of male rats by inhibiting promoter methylation of imprinted gene H19 by down-regulating DNMT3a

作　　者：谢明仁 何天晓 王贤贤 谢刘阳 袁霞 张璟 XIE Ming-ren;HE Tian-xiao;WANG Xian-xian;XIE Liu-yang;YUAN Xia;ZHANG Jing(Public Security Branch,Gansu University of Political Science and Law,Key Laboratory of Evidence of Science and Technology Research and Application,Lanzhou 730070,China)

机构地区：[1]甘肃政法大学公安分院,甘肃省证据科学技术研究与应用重点实验室,甘肃兰州730070

出　　处：《中国药理学通报》2023年第8期1515-1521,共7页Chinese Pharmacological Bulletin

基　　金：甘肃省科技计划项目(No 21JR7RA571);甘肃政法大学项目(No GZF2021XZD06)。

摘　　要：目的探讨F0代雄鼠吸食海洛因对F1代大鼠成瘾与复吸的影响及机制。方法用不同剂量(1、3、9 mg·kg^(-1))海洛因处理F0代雄鼠。Western blot检测F0代雄鼠精子和F1代大鼠背侧海马组织中DNA甲基转移酶3a(DNMT3a)蛋白表达水平。DNA甲基化免疫共沉淀(MeDIP)-qRT-PCR检测H19启动子甲基化率。实时荧光定量PCR(qRT-PCR)检测H19 mRNA表达水平。构建海洛因自身给药模型,检测F1代各组大鼠的成瘾与复吸行为。结果海洛因明显降低F0代雄鼠精子中DNMT3a蛋白表达水平及H19启动子甲基化率,明显提高H19 mRNA表达水平(P<0.05)。F0代雄鼠吸食海洛因明显降低F1代大鼠背侧海马组织中H19启动子甲基化率,明显提高H19 mRNA表达水平(P<0.05)。F0代雄鼠吸食海洛因明显增加F1代大鼠在海洛因自身给药实验和复吸实验中的有效鼻触次数(P<0.05)。结论F0代雄鼠吸食海洛因会促进F1代大鼠对海洛因的成瘾与复吸,其机制可能与海洛因通过下调F0代雄鼠精子中DNMT3a表达抑制印记基因H19启动子甲基化,进而促进F1代大鼠背侧海马组织中H19 mRNA表达有关。Aim To investigate the effect of heroin use in male rats of F0 generation on heroin addiction and relapse in rats of F1 generation and the underlying mechanism.Methods Male rats of F0 generation were treated with different doses of heroin(1,3,9 mg·kg^(-1)).Western blot was used to detect the expression of DNA methyltransferase 3a(DNMT3a)protein in sperm of male rats of F0 generation and dorsal hippocampal tissue of F1 generation.DNA methylation immunoprecipitation(MeDIP)-qRT-PCR was used to detect the methylation rate of H19 promoter.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the expression level of H19 mRNA.Heroin self-administration model was established.Addiction and relapse behaviors of the rats of F1 generation were detected.Results Heroin significantly decreased DNMT3a protein expression level and H19 promoter methylation rate in sperm of male rats of F0 generation,and significantly increased H19 mRNA expression level(P<0.05).Heroin use in male rats of F0 generation significantly decreased the methylation rate of H19 promoter and increased the expression level of H19 mRNA in the dorsal hippocampus of rats of F1 generation(P<0.05).Heroin use in male rats of F0 generation significantly increased the number of responses to active holes of rats of F1 generation during heroin self-administration experiments and relapse experiments(P<0.05).Conclusions Heroin use in male rats of F0 generation can promote heroin addiction and relapse in rats of F1 generation.The mechanism may be related to the promotion of H19 mRNA expression in dorsal hippocampus of rats of F1 generation by inhibiting the methylation of the promoter of H19 by down-regulating the expression of DNMT3a of sperm of male rats of F0 generation.

关 键 词：海洛因 DNMT3a H19 甲基化 成瘾 复吸 

分 类 号：R-332[医药卫生] R394.2R749.61R971.2