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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,进而促进HIV-1的感染复制。
To investigate the effect of Opiates on TLR9 gene expression in peripheral blood mononuclear cells (PBMCs) infected with HIV-1 and to explore the role of opioid drugs in promoting HIV-1 replication The mechanism of action to lay the foundation. HIV-1 infected patients (Opiates HIV (+) group) were enrolled in the methadone and AIDS voluntary counseling and testing clinics in Nanning, Liuzhou and Qinzhou districts. After informed consent, , Opioid abuse non-HIV-1 infection (Opiates HIV (-)), non-opiates HIV (non-opiates HIV) and healthy control (non-opiates HIV) Randomly recruited 50 people. Secondly, we investigated the demographic characteristics of the subjects by questionnaires and collected peripheral venous blood. PBMCs were isolated and RNA was extracted. Finally, qPCR and Western blot (WB) were used to detect the mRNA and protein expression of TLR9 in PBMCs of four groups. The survey found that the four groups of people in terms of age, gender, ethnicity, residence location, marital status, education level and the age of drug abuse and other demographic characteristics were not statistically significant (P> 0.05). The median viral load of Opiates HIV (+) and Non-Opiates HIV (+) were 4.450 × 103 and 3.977 × 103 cp / mL, respectively, with statistical significance (P <0.05). The relative expression of TLR9 mRNA was (2.13 ± 1.59) × 10-3 and (3.66 ± 2.22) × 10-3 in Opiates HIV (+), Non-Opiates HIV (+) and Opiates HIV , (1.96 ± 1.42) × 10-3 and (7.66 ± 4.87) × 10-3, respectively. There was interaction between opioid abuse and HIV infection on TLR9 expression (F = 25.91, P = 0.000). Independent effect analysis revealed that the relative expression levels of TLR9 in opioid abusers were significantly lower than those in non-drug abusers (P <0.05) in HIV-positive and -negative populations. Among drug users, HIV-positive and HIV- There was no significant difference between the two groups (P> 0.05). Among the non-drug users, the positive rate of HIV was lower than that of the normal group. There was significant difference between the two groups (P <0.05). The results of WB showed that the expression of TLR9 protein in Opiates HIV (+), Non-Opiates HIV (+) and Opiates HIV (-) groups was lower than that in Control group. The above results indicate that opioid drugs can down-regulate TLR9 expression in PBMCs infected with HIV-1, suggesting that opioid drugs may promote the replication of HIV-1 infection by affecting TLR9-mediated immune response.