摘要
目的通过实时荧光定量PCR（RT-PCR）的方法检测中国HIV／AIDS患者外周血单个核细胞（PBMC）中TLR4 mRNA的表达水平，探讨TLR4 mRNA表达水平与HIV感染后疾病进展的关系。方法构建pUCm-TLR4质粒作为定量模板，在LightCycler检测仪上测定62例HIV／AIDS患者和30名健康对照者PBMC中TLR4 mRNA的含量。结果（1）HIV／AIDS患者PBMC中TLR4 mRNA的表达范围是（4．0×10^5-1．04×10^8）拷贝／μg RNA，显著高于健康对照组的（4．5×10^5-1．25×10^6）拷贝／μg RNA，（P〈0．01）。（2）在HIV／AIDS患者组中，艾滋病（AIDS）组外周血中，TLR4 mRNA的含量高于HIV慢性感染（HIV）组及长期不进展（LTNP）组，HIV组高于LTNP组（P〈0．05），LTNP组与对照组比较差别无统计学意义。（3）HIV／AIDS患者外周血中TLR4 mRNA的含量与HIV-1RNA病毒载量呈显著正相关（r=0．697，P〈0．01）。结论成功建立了人TLR4 mRNA表达水平的荧光定量检测方法；中国HIV／AIDS患者外周血单个核细胞中TLR4 mRNA的含量显著高于健康对照组，且与疾病进展密切相关。
Objective To detect the expression of TLR4 mRNA in peripheral blood mononuclear cells（PBMC） of Chinese HIV/AIDS patients through real-time quantitative reverse transcription -polymerase chain reaction（RT-PCR） and explore the relationship between expression of TLR4 mRNA and HIV disease progression. Methods pUCm-TLR4 plasmid was constructed as the quantitative template and Light Cycler was used to detect the expression of TLR4 mRNA in PBMC of 60 HIV/AIDS patients and 30 healthy control. Results （ 1 ） The expression of TLR4 mRNA in PBMC of Chinese HIV/AIDS patients ranges from 4. 0 × 10^5 to 10. 4 × 10^7 copy/lag RNA, significantly higher than the group of healthy control （4. 5×10^5 - 1.25×10^6 copy/lag RNA,P 〈0.01 ）. （2） In the group of HIV/AIDS patients, the expression of TLR4 mRNA in the group of AIDS is higher than the group of chronic HIV and the group of long-term nonprogressors（LTNP）. The expression in the group of chronic HIV is higher than the group of LTNP （ P 〈 0.05 ）. There was no significant difference between the group of LTNP and the group of healthy control. （ 3 ） The expression of TLR4 mRNA in PBMC of Chinese HIV/AIDS patients is positively correlated with HIV-1 RNA viral load （ r = 0. 697, P 〈 0. 01 ）. Conclusions We successfully constructed the fluorescence quantitative method to detect the expression of TLR4 mRNA in human. The expression of TLR4 mRNA in PBMC of Chinese HIV/AIDS patients is significantly higher than the group of healthy control, and the expression is closely correlated with the disease progression.
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2007年第9期1005-1008,共4页
Chinese Journal of Laboratory Medicine
基金
国家自然科学基金(30471548)
辽宁省优秀青年科研人才培养资金资助项目(3040011)
国家973重点基础研究发展计划资助项目(2006CB504206)
辽宁省教育厅高等学校科研项目(05L548)
