黄芩苷对人淋巴细胞Toll样受体的调节

发布时间：2018-05-20 07:56

本文选题：黄芩苷 + T淋巴细胞　；参考：《南昌大学》2008年硕士论文

【摘要】： 目的:研究黄芩苷对人淋巴细胞Toll样受体(TLR)的调节,探讨其在免疫调节方面的作用。方法: 1.采用尼龙毛分离法分离人外周血T和B细胞;2.采用UF-50流式分析寻找最佳黄芩苷作用浓度;3.采用实时定量荧光逆转录聚合酶链反应(RT-PCR),分析在最佳黄芩苷作用浓度处理前后TLRs的表达情况;4.通过受体抑制实验和实时定量荧光RT-PCR探讨黄芩苷对TLRs的调节位点。结果:1.体外孵育48h后,1mg/ml的黄芩苷可以显著促进人T、B淋巴细胞增殖(P0.05);2.使用1mg/ml黄芩苷相对于未用黄芩苷处理的T、B细胞,其TLR3、TLR7、TLR8及TLR9mRNA的表达均有显著增加(P0.05),T细胞增加倍数分别为21、15、57和66倍,B细胞增加的倍数分别为24、20、61和63倍,TLR1、TLR2、TLR4、TLR5、TLR6及TLR10 mRNA表达变化不显著(P0.05);3.在1mg/ml黄芩苷作用下人T、B细胞TLR3、7、8和9 mRNA在12h时表达已开始增加,除T细胞的TLR7 mRNA表达在48h时略有下降外,其余表达基本一直持续增加,至48h到达峰值;4.人T、B细胞在被黄芩苷作用前使用鼠抗人TLR4单克隆抗体封闭,会显著降低黄芩苷对TLR3、7、8和9 mRNA表达的诱导作用(P0.05),使得T细胞增加倍数分别降为11、5、28和36倍,下降比率分别为47.6%、66.7%、50.9%和45.5%;B细胞增加的倍数分别降为12、11、30和31倍,下降比率分别为50.0%、45.0%、50.8%和50.8%。结论:1.黄芩苷在体外能显著促进T、B淋巴细胞增殖;2.人外周血T、B细胞组成性表达含量差异较大的TLR1-10mRNA,黄芩苷在体外可以显著上调人T、B淋巴细胞TLR3、TLR7、TLR8及TLR9 mRNA的表达,但对TLR1、TLR2、TLR4、TLR5、TLR6及TLR10 mRNA没有影响;3.黄芩苷可能通过TLR4参与对人天然免疫能力的调节。
[Abstract]:Aim: to study the regulation of baicalin on Toll like receptor (TLR) in human lymphocytes and its role in immunomodulation. Methods: 1. Human peripheral blood T and B cells were separated by nylon wool method. UF-50 flow analysis was used to find the best concentration of baicalin. Real-time quantitative fluorescence reverse transcriptase polymerase chain reaction (RT-PCR) was used to analyze the expression of TLRs before and after the best concentration of baicalin. The regulatory sites of baicalin on TLRs were investigated by receptor inhibition test and real-time quantitative RT-PCR. The result is 1: 1. After incubation in vitro for 48 h, baicalin (1 mg / ml) could significantly promote the proliferation of human TnB lymphocytes. The expression of TLR8 and TLR9mRNA in 1mg/ml baicalin was significantly higher than that in TLB cells without baicalin treatment. The expression of TLR8 and TLR9mRNA in TLR3, TLR7, TLR7, TLR8, TLR8, TLR7, TLR8, TLR7, TLR8, TLR7, TLR8, TLR8, TLR8 and TLR6 of TLR7, TLR7, TLR6, and TLR6 in TLR7, TLR7, TLR6, TLR7, TLR6, TLR7, TLR6, TLR6, TLR7, TLR6, and TLR6, The expression of TLR3O78 and 9 mRNA began to increase at 12h after the action of 1mg/ml baicalin, and the expression of TLR3n78 and 9 mRNA increased continuously at 48h, except that the TLR7 mRNA expression of T cells decreased slightly at 48h. The blocking of mouse anti-human TLR4 monoclonal antibody before being treated with baicalin could significantly reduce the induction of baicalin on the expression of TLR3O7O8 and 9 mRNA, and reduce the T cell proliferation multiple to 115mand 36-fold, respectively. The decreasing rates were 47.6% and 45.5%, respectively, and the increase times of B cells were decreased to 120.1130 and 31 times, respectively, and the decreasing rates were 50.0% and 50.8%, respectively. Conclusion 1. Baicalin can significantly promote the proliferation of T B lymphocytes in vitro. In vitro, baicalin could significantly up-regulate the expression of TLR3, TLR7, TLR8 and TLR9 mRNA in human peripheral blood TLR1-10mRNAs, but had no effect on TLR1-10mRNAs and TLR5TLR6 and TLR10 mRNA. Baicalin may be involved in the regulation of human innate immunity through TLR4.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392

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