阿片类毒品对HIV-1感染者PBMCs中TLR9基因表达的影响  

Influence of Opiate Abuse on Expression of Toll-like Receptor 9 in Peripheral Blood Mononuclear Cells of HIV-1-Infected Individuals

作　　者：潘沛江 韦富梅[1] 蒋俊俊[1] 梁冰玉[1] 黄颉刚[1] 廖艳研[2] 苏锦明[1] 李彧[1] 杨小艺[1] 陈晖[3] 叶力[1] 梁浩[1] 

机构地区：[1]广西医科大学公共卫生学院,广西艾滋病防治研究重点实验室,南宁530021 [2]广西壮族自治区疾病预防控制中心消杀与媒介控制所,南宁530028 [3]广西医科大学第一附属医院老年消化内科,南宁530021

出　　处：《病毒学报》2015年第2期132-138,共7页Chinese Journal of Virology

基　　金：国家自然科学基金资助项目(81171624);教育部博导课题(20114503110003);广西医科大学青年科学基金项目(GXMUYSF201210)

摘　　要：探讨滥用阿片类毒品(Opiates)对Ⅰ型艾滋病病毒(Human immunodeficiency virus,HIV-1)感染者外周血单个核细胞(PBMCs)中TLR9基因表达水平的影响,为阐明阿片类毒品促进HIV-1复制的作用机制奠定基础。首先在南宁、柳州、钦州市地区的美沙酮、艾滋病自愿咨询检测门诊招募对象,经研究对象知情同意,分为4组,即阿片类毒品滥用的HIV-1感染组(Opiates HIV(+)组)、阿片类毒品滥用非HIV-1感染组(Opiates HIV(-)组)、非阿片类毒品滥用的HIV感染组(Non-opiates HIV(+)组)和健康对照组(Control组),每组随机招募50人。其次,以问卷形式调查对象的人口学特征,并采集其外周静脉血,分离出PBMCs后提取RNA。最后采用qPCR、蛋白质印迹法(Western blot,WB)法检测4组人群PBMCs中TLR9的mRNA、蛋白表达水平。经调查发现4组人群在年龄、性别、民族、户籍所在地、婚姻状况、文化程度和吸毒年限等人口学特征方面差异均无统计学意义(P>0.05)。Opiates HIV(+)与Non-Opiates HIV(+)组病毒载量中位数分别为4.450×103和3.977×103 cp/mL,差异有统计学意义(P<0.05)。TLR9 mRNA相对表达量在Opiates HIV(+)、Non-Opiates HIV(+)、Opiates HIV(-)和Control组中分别为(2.13±1.59)×10-3、(3.66±2.22)×10-3、(1.96±1.42)×10-3和(7.66±4.87)×10-3。阿片类毒品滥用和HIV感染对TLR9的表达存在交互作用(F=25.91,P=0.000)。经单独效应分析,HIV阳性与阴性人群中,阿片类毒品滥用者TLR9相对表达量均明显低于非毒品滥用者(P<0.05);在吸毒人群中,HIV阳性者与HIV阴性者两组间差异没有统计学意义(P>0.05);在不吸毒人群中,HIV阳性者低于正常组,两组间差异有统计学意义(P<0.05)。WB的结果显示,Opiates HIV(+)、Non-Opiates HIV(+)、Opiates HIV(-)三组TLR9蛋白的表达量均低于Control组。上述结果表明阿片类毒品能够下调HIV-1感染者PBMCs中TLR9的表达水平,提示阿片类毒品可能通过影响TLR9所介导的机体免疫效应,进而促�The aim of this study was to investigate the influence of opiate abuse on the expression of Tolllike receptor 9（TLR9）in the peripheral blood mononuclear cells（PBMCs）of HIV-1-infected patients and to elucidate possible mechanisms involved in the enhancement of HIV-1replication by opiate abuse.A total of 200 participants were enrolled in the study by random selection from methadone treatment centers and voluntary HIV counseling and testing centers in the cities of Nanning,Liuzhou,and Qinzhou.These participants included 50HIV-positive opiate abusers（Opiates HIV（＋）group）,50HIV-negative opiate abusers（Opiates HIV（-）group）,50 HIV-positive subjects who were not opiate abusers（Non-opiates HIV（＋）group）,and 50HIV-negative subjects who were not opiate abusers（Control group）.PBMCs were isolated from the peripheral blood samples from the subjects and the expression levels of TLR9 mRNA and protein were determined by q-PCR and western blot respectively.There was no significant difference among the four groups in age,gender,nationality,domicile,marital status,educational background or duration of drug abuse（P0.05）.The median viral loads of the Opiates HIV（＋）were significantly higher than those of the Non-Opiates HIV（＋）groups（4.450×103 and 3.977×103 copies/mL respectively,P0.05）.The relative expression levels of TLR9 mRNA in the Opiates HIV（＋）,Non-Opiates HIV（＋）,Opiates HIV（-）and Control groups were（2.13±1.59）×10-3,（3.66±2.22）×10-3,（1.96±1.42）× 10-3 and（7.66 ± 4.87）× 10-3,respectively.The expression of TLR9 mRNA was significantly lower in both HIV-1-infected and-uninfected groups of opiate abusers compared with groups of non-abusers（P0.05）.There was no significant difference in TLR9 mRNA expression levels between the Opiates HIV（＋）group and the Opiates HIV（-）group（P0.05）.However,in the non-opiate groups,the expression levels of TLR9 mRNA in the HIV（＋）group were significantly lower than that of the control

关 键 词：阿片类毒品 HIV-1 TLR9受体 

分 类 号：R593.3[医药卫生—内科学]