龙胆泻肝配方颗粒治疗生殖器疱疹的临床观察及其对外周血树突状细胞功能影响的研究
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摘要
目的：观察龙胆泻肝配方颗粒治疗湿热型生殖器疱疹的临床疗效；通过对患者治疗前后外周血树突状细胞功能变化的实验研究,来探讨中药龙胆泻肝配方颗粒抗病毒的免疫作用机制。
方法：1)将60例生殖器疱疹患者随机分为龙胆泻肝配方颗粒组、龙胆泻肝汤方组及西药阿昔洛韦组,观察治疗前后患者临床证候积分,判断其近期临床疗效。并且记录患者治疗后6个月内的复发情况,判断其远期临床疗效。2)在龙胆泻肝配方颗粒治疗组患者治疗前后,分别抽取静脉血10ml,及健康志愿者静脉血10ml。用淋巴细胞分离液分离外周血单个核细胞,体外经GM-CSF.IL-4,TNF-α的诱导生成大量的成熟树突状细胞,倒置显微镜观察其形态变化,流式细胞仪检测细胞表面CD11c、CD80、CD83、CD86、HLA-DR和HLA-ABC的表达。3)采用荧光定量RT-PCR的方法检测了治疗组患者治疗前后外周血树突状细胞TLR1～TLR9mRNA的表达情况。
结果：1)龙胆泻肝汤与龙胆泻肝配方颗粒治疗湿热下注型生殖器疱疹的近期疗效与远期疗效均优于西药阿昔洛韦片。龙胆泻肝配方颗粒的疗效与传统汤药的疗效相当。2)生殖器疱疹患者外周血树突状细胞表面的免疫分子,CD11c、CD80、CD83、CD86、HLA-DR和HLA-ABC的表达水平,均较正常人增高(P<0.05)。经龙胆泻肝配方颗粒治疗后,患者外周血树突状细胞表面的免疫分子的表达水平趋向于正常(P>0.05)。3)治疗前患者外周血树突状细胞TLR1～TLR9mRNA的表达水平与健康对照组比较,除TLR2mRNA外,均有统计学差异(P<0.05),其中TLR3、TLR4、TLR7、TLR8、TLR9mRNA的表达较正常水平升高；TLR1、TLR5、TLR6mRNA的表达较正常水平降低；TLR2mRNA的表达较正常水平无明显差异(P>0.05)。治疗后,患者外周血树突状细胞TLR1-TLR9mRNA的表达水平均较治疗前倾向于正常(P>0.05)。
结论：龙胆泻肝配方颗粒治疗湿热下注型生殖器疱疹有确切的临床疗效。龙胆泻肝配方颗粒能调节生殖器疱疹患者外周血Toll样受体的表达水平,通过特异的信号传导通路诱导树突状细胞表达高水平的辅助刺激因子,抗原提呈功能增强,产生炎性细胞因子和趋化因子,诱导生成病毒特异性T细胞,分泌细胞毒素杀伤靶细胞,促进机体发挥抗病毒的作用。
Objective To investigate the clinical significance of Longdanxiegan Chinese traditional medicine Granule in preventing and curing the Shire type genital herpes.To explore the immunological function of Longdanxiegan Granule,through studying the Dendritic cells extracted from the patients'blood.
Methods 1) The 60 patients were randomly divided into 3 groups completely.20 were treated with Aciclovir.20 were with Longdanxiegan Chinese traditional medicine Granule.20 were with Longdanxiegan Chinese traditional medicine. Observed the integral of clinical symptoms, clinical efficient rate, and the recurring rate respectively before and after the treatment.2) Peripheral blood mononuclear cells (PBMC) were separated from heparinized venous blood by stimulating factor GM-CSF、IL-4、TNF-α. The Dendritic cells were harvested for phenotypic analysis by flow cytometry.Then some phenotype changes such as CD11c、CD80、CD83、CD86、HLA-DR and HLA-ABC of DCs were observed.3) RNA Of the dendritic cells was extracted by Trizol. The levels of mRNA for TLR1-TLR9 were examined by fluorescent quantitative RT-PCR.
Result 1) The integral of clinical symptoms were improved obviously after theatment for three groups.There was no significant difference in clinical rate for the two Chinese traditional medicine groups. The group treated with Longdanxiegan Chinese traditional medicine Granule showed a lower recurring rate than the Aciclovir group.2) The expression of CD11c、CD80、CD83、CD86、HLA-DR and HLA-ABC on DCs were higher in patients than that in healthy controls(P＜0.05). After theatment, the expression of these molecules were regulated, there were no significant differences between patients and controls(P＞0.05).3) The expressions of TLR3、TLR4、TLR7、TLR8 and TLR9 mRNA were significantly higher in patients than that in healthy controls(P<0.05);but the expressions of TLR1、TLR5 and TLR6 mRNA were markedly lower than that in controls(P＜0.05).After theatment, there were no significant differences in the expressions of the TLR1-TLR9mRNA between patients and controls(P＞0.05).
Conclusion Longdanxiegan Chinese traditional medicine Granule is effective for the Shire type genital herpes. Longdanxiegan granule could adjust the expressions of TLRs on peripheral blood leucocyte in patients with genital herpes and play a key role in initiating innate immune responses to viral pathogens.
方法：1)将60例生殖器疱疹患者随机分为龙胆泻肝配方颗粒组、龙胆泻肝汤方组及西药阿昔洛韦组,观察治疗前后患者临床证候积分,判断其近期临床疗效。并且记录患者治疗后6个月内的复发情况,判断其远期临床疗效。2)在龙胆泻肝配方颗粒治疗组患者治疗前后,分别抽取静脉血10ml,及健康志愿者静脉血10ml。用淋巴细胞分离液分离外周血单个核细胞,体外经GM-CSF.IL-4,TNF-α的诱导生成大量的成熟树突状细胞,倒置显微镜观察其形态变化,流式细胞仪检测细胞表面CD11c、CD80、CD83、CD86、HLA-DR和HLA-ABC的表达。3)采用荧光定量RT-PCR的方法检测了治疗组患者治疗前后外周血树突状细胞TLR1～TLR9mRNA的表达情况。
结果：1)龙胆泻肝汤与龙胆泻肝配方颗粒治疗湿热下注型生殖器疱疹的近期疗效与远期疗效均优于西药阿昔洛韦片。龙胆泻肝配方颗粒的疗效与传统汤药的疗效相当。2)生殖器疱疹患者外周血树突状细胞表面的免疫分子,CD11c、CD80、CD83、CD86、HLA-DR和HLA-ABC的表达水平,均较正常人增高(P<0.05)。经龙胆泻肝配方颗粒治疗后,患者外周血树突状细胞表面的免疫分子的表达水平趋向于正常(P>0.05)。3)治疗前患者外周血树突状细胞TLR1～TLR9mRNA的表达水平与健康对照组比较,除TLR2mRNA外,均有统计学差异(P<0.05),其中TLR3、TLR4、TLR7、TLR8、TLR9mRNA的表达较正常水平升高；TLR1、TLR5、TLR6mRNA的表达较正常水平降低；TLR2mRNA的表达较正常水平无明显差异(P>0.05)。治疗后,患者外周血树突状细胞TLR1-TLR9mRNA的表达水平均较治疗前倾向于正常(P>0.05)。
结论：龙胆泻肝配方颗粒治疗湿热下注型生殖器疱疹有确切的临床疗效。龙胆泻肝配方颗粒能调节生殖器疱疹患者外周血Toll样受体的表达水平,通过特异的信号传导通路诱导树突状细胞表达高水平的辅助刺激因子,抗原提呈功能增强,产生炎性细胞因子和趋化因子,诱导生成病毒特异性T细胞,分泌细胞毒素杀伤靶细胞,促进机体发挥抗病毒的作用。
Objective To investigate the clinical significance of Longdanxiegan Chinese traditional medicine Granule in preventing and curing the Shire type genital herpes.To explore the immunological function of Longdanxiegan Granule,through studying the Dendritic cells extracted from the patients'blood.
Methods 1) The 60 patients were randomly divided into 3 groups completely.20 were treated with Aciclovir.20 were with Longdanxiegan Chinese traditional medicine Granule.20 were with Longdanxiegan Chinese traditional medicine. Observed the integral of clinical symptoms, clinical efficient rate, and the recurring rate respectively before and after the treatment.2) Peripheral blood mononuclear cells (PBMC) were separated from heparinized venous blood by stimulating factor GM-CSF、IL-4、TNF-α. The Dendritic cells were harvested for phenotypic analysis by flow cytometry.Then some phenotype changes such as CD11c、CD80、CD83、CD86、HLA-DR and HLA-ABC of DCs were observed.3) RNA Of the dendritic cells was extracted by Trizol. The levels of mRNA for TLR1-TLR9 were examined by fluorescent quantitative RT-PCR.
Result 1) The integral of clinical symptoms were improved obviously after theatment for three groups.There was no significant difference in clinical rate for the two Chinese traditional medicine groups. The group treated with Longdanxiegan Chinese traditional medicine Granule showed a lower recurring rate than the Aciclovir group.2) The expression of CD11c、CD80、CD83、CD86、HLA-DR and HLA-ABC on DCs were higher in patients than that in healthy controls(P＜0.05). After theatment, the expression of these molecules were regulated, there were no significant differences between patients and controls(P＞0.05).3) The expressions of TLR3、TLR4、TLR7、TLR8 and TLR9 mRNA were significantly higher in patients than that in healthy controls(P<0.05);but the expressions of TLR1、TLR5 and TLR6 mRNA were markedly lower than that in controls(P＜0.05).After theatment, there were no significant differences in the expressions of the TLR1-TLR9mRNA between patients and controls(P＞0.05).
Conclusion Longdanxiegan Chinese traditional medicine Granule is effective for the Shire type genital herpes. Longdanxiegan granule could adjust the expressions of TLRs on peripheral blood leucocyte in patients with genital herpes and play a key role in initiating innate immune responses to viral pathogens.
引文
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