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Methamphetamine Inhibits Toll Like Receptor-9-Mediated Activation Anti-HIV Activity in Macrophages

Author: ZuoPing
Tutor: LiangHao
School: Guangxi Medical University
Course: Epidemiology and Biostatistics,
Keywords: Ice HIV TLR-9 CpG-ODN
CLC: R114
Type: Master's thesis
Year: 2010
Downloads: 42
Quote: 1
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Abstract


The first part: ice TLR-9 mediated anti-HIV activity Objective: To observe the effects of methamphetamine on the specificity of TLR-9 ligand CpG-ODN inhibition of macrophages infected with HIV produce. Methods: Fresh peripheral blood collected from healthy adults, mononuclear cells were isolated, and then by purified human macrophages were cultured adherent. Human macrophages with ice and / or dopamine receptor D1 blockers · (SCH23390), CpG-ODN pretreatment, and then with HIV Bal virus-infected cells, and after 4, 6, 8, 10, respectively, in the exposure, 12 days culture supernatant was collected for the detection of HIV reverse transcriptase activity, and observed under the microscope with the ice impact of HIV infection in the role of CpG-ODN on human macrophages. Results: 1. Methamphetamine HIV reverse transcriptase activity of the treatment groups were higher than the blank group 2.ODN treatment group of HIV reverse transcriptase activity detection value and the blank control group and significantly reduced compared. 3 of ice / ODN treatment group alone ODN treatment group compared to HIV reverse transcriptase activity was significantly increased. Ice / ODN / SCH / treated HIV reverse transcriptase activity was significantly lower than that of ice / ODN group. Conclusion: CpG-ODN TLR-9 receptor inhibition of human macrophages of HIV-1 infection, the ice can weaken the TLR-9 inhibition of HIV-infected macrophages. 2 ice may be acting at the dopamine receptor D1 weakened the role of TLR-9 mediated resistance to HIV infection. The second part: methamphetamine inhibit the TLR-9 mediated anti-HIV mechanism research purposes: to observe the ice TLR-9 in human macrophages and IFN-α gene expression, TLR-9 mediated antiviral IRF-7 signal transduction pathways, MyD88, the role of the expression of MxA, ISG56 immune factors, to investigate methamphetamine weakened TLR-9 inhibit HIV replication mechanisms. Methods: Fresh peripheral blood collected from healthy adults, mononuclear cells were isolated, and then by purified human macrophages were cultured adherent. Time gradient and concentration gradient method, adding ice cultured cells using real-time quantitative PCR to detect cell TLR-9 and endogenous IFN-α gene mRNA levels. Meanwhile, fluorescence immunoassay and ELISA methods were used to detect expression of TLR-9 and endogenous IFN-α protein in macrophages. In addition, the use of ice and / or dopamine receptor D1 blocker (SCH23390) and CpG-ODN on human macrophages pretreatment by real-time quantitative PCR detection TLR-9 mediated antiviral signal transduction pathways, IRF-7 , MyD88, MxA, ISG56 mRNA expression levels. Results: 1. Methamphetamine treated TLR-9 gene expression was significantly decreased with the the exposed amount and the number of days and gradually reduce the dose - and time - effect relationship (P = lt; 0.01). Ice / ODN treated TLR-9 gene expression was significantly lower than the ODN treated; methamphetamine / ODN / SCH treated TLR-9 gene expression above the ice / ODN group (P = lt; 0.01). 3. The methamphetamine treated with IFN-α gene expression was significantly decreased, and with the added amount of methamphetamine and dealing with the number of days gradually reduce the dose - and time - effect relationship (P = lt; 0.05). Ice / ODN treated with IFN-α gene expression was significantly lower than the ODN treated; the ice / ODN / SCH treated with IFN-α gene expression was significantly higher than the ice / ODN group (P = lt; 0.05). 5.ODN treatment group compared with the blank control group, IRF7, MyD88 and MxA expression were significantly increased (P lt; 0.05), no significant difference in ISG56 gene expression blank control group; ice / ODN treated IRF7, MyD88, MxA ISG56 gene expression was significantly below ODN treated group (P lt; 0.05); join dopamine receptor antagonist pretreatment, ice / ODN / SCH treated MyD88 and ISG56 gene expression with ice / ODN2216 group of significantly increased compared (P lt; 0.05), IRF7 and MxA gene expression did not change significantly. Conclusion: 1. Methamphetamine may TLR-9 expression by inhibiting macrophage down the generation of endogenous IFN-α. 2 ice can significantly weaken the CpG-ODN-mediated cell expression of TLR-9 and IFN-α. Ice can cut TLR-9 Dui IRF7, MyD88, of MxA and ISG56 cytokines induced .4. Methamphetamine weaken the role of TLR-9 expression regulating immune cytokines may be blocking dopamine receptors D1 blockers.

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