斑秃患者PBMC中Th1/Th2型细胞因子、转录因子T-bet mRNA表达状况及中药对其调节作用
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摘要
目的
探讨肝肾不足型斑秃患者外周血单一核细胞(PBMC)中Th1/Th2型细胞因子、转录因子T-bet mRNA表达状况与斑秃中医证型及发病的关系;中药原花青素(PC)、黄芪多糖(APS)对斑秃患者PBMC中Th1/Th2型细胞因子、T-bet基因表达的调节作用。
方法
用逆转录-聚合酶链反应(RT-PCR)分别检测8例轻型斑秃、12例重型斑秃患者及10例正常人经PHA刺激后PBMC中Th1型细胞因子(IFN-γ、IL-12)、Th2型细胞因子IL-10及转录因子T-bet mRNA的表达水平;同时还检测经PC、APS分别与PHA共同刺激斑秃患者PBMC后Th1/Th2型细胞因子、转录因子T-bet mRNA表达水平的改变情况。
结果
经PHA刺激后轻型斑秃患者PBMC中Tb1型细胞因子(IFN-γ、IL-12)基因表达水平轻度升高,与正常对照比较无显著性差异(P>0.05);Th2型细胞因子(IL-10)基因表达下降,显著低于正常对照组(P<0.05);而转录因子T-betmRNA表达水平明显升高,显著高于正常对照组(P<0.05)。经PHA刺激后重型斑秃患者PBMC中Th1型细胞因子(IFN-γ、IL-12)及转录因子T-bet基因表达不仅显著高于正常对照组(P<0.01),而且还高于轻型斑秃病人(P<0.05);Th2型细胞因子(IL-10)基因表达显著低于正常对照组(P<0.05),与轻型斑秃病人比较无显著性差异。PC、APS分别与PHA共同刺激斑秃患者PBMC后可抑制Th1型细胞因子(IFN-γ、IL-12)及转录因子T-bet基因表达,逆转斑秃病人Th1型反应。相关性分析表明:转录因子T-bet与IFN-γ呈高度正相关(R=0.806,P<0.001);转录因子T-bet与IL-12呈高度
Objectives First to investigate the relationship between expression of Thl/Th2 types cytokines(IFN- γ、 IL-12/IL-10) & transcription factor T-bet mRNA in PBMC of the alopecia areata patients with symptoms of liver-kiney deficiency and TCM symptoms; Second to study the role of expression of Thl/Th2 types cytokines & transcription factor T-bet mRNA in PBMC in the pathogenesis of alopecia areata and its regulation by Procyanidins(PC) and Astraglus polysaccharide(APS).Methods Apply RT-PCR technique to detect the mRNA expression of Thl/Th2 types cytokines & transcription factor T-bet in PBMC of the patients with alopecia areata by stimulation with phytohemagglutinin(PHA) and PHA plus PC/APS respectively.Results The mRNA expression of Thl type cytokines in PHA-stimulated PBMC of the patients with localized alopecia areata were increased slightly compared with healthy controls, the expression of transcription factor T-bet mRNA markedly elevated with the healthy controls (p<0. 05) , on the contrary, the mRNA expression of Th2 type cytokines (IL-10)lower significantly than healthy controls(P<0. 05). However, the mRNA expression of both Thltype cytokines & transcription factor T-bet in PHA-stimulated PBMC of the patients with extensive alopecia areata were markedly increased compared with healthy controls(P<0. 01)and localized form(P<0. 05), but the mRNA expression of Th2 type cytokines (IL-10)were lower significantly than healthy
探讨肝肾不足型斑秃患者外周血单一核细胞(PBMC)中Th1/Th2型细胞因子、转录因子T-bet mRNA表达状况与斑秃中医证型及发病的关系;中药原花青素(PC)、黄芪多糖(APS)对斑秃患者PBMC中Th1/Th2型细胞因子、T-bet基因表达的调节作用。
方法
用逆转录-聚合酶链反应(RT-PCR)分别检测8例轻型斑秃、12例重型斑秃患者及10例正常人经PHA刺激后PBMC中Th1型细胞因子(IFN-γ、IL-12)、Th2型细胞因子IL-10及转录因子T-bet mRNA的表达水平;同时还检测经PC、APS分别与PHA共同刺激斑秃患者PBMC后Th1/Th2型细胞因子、转录因子T-bet mRNA表达水平的改变情况。
结果
经PHA刺激后轻型斑秃患者PBMC中Tb1型细胞因子(IFN-γ、IL-12)基因表达水平轻度升高,与正常对照比较无显著性差异(P>0.05);Th2型细胞因子(IL-10)基因表达下降,显著低于正常对照组(P<0.05);而转录因子T-betmRNA表达水平明显升高,显著高于正常对照组(P<0.05)。经PHA刺激后重型斑秃患者PBMC中Th1型细胞因子(IFN-γ、IL-12)及转录因子T-bet基因表达不仅显著高于正常对照组(P<0.01),而且还高于轻型斑秃病人(P<0.05);Th2型细胞因子(IL-10)基因表达显著低于正常对照组(P<0.05),与轻型斑秃病人比较无显著性差异。PC、APS分别与PHA共同刺激斑秃患者PBMC后可抑制Th1型细胞因子(IFN-γ、IL-12)及转录因子T-bet基因表达,逆转斑秃病人Th1型反应。相关性分析表明:转录因子T-bet与IFN-γ呈高度正相关(R=0.806,P<0.001);转录因子T-bet与IL-12呈高度
Objectives First to investigate the relationship between expression of Thl/Th2 types cytokines(IFN- γ、 IL-12/IL-10) & transcription factor T-bet mRNA in PBMC of the alopecia areata patients with symptoms of liver-kiney deficiency and TCM symptoms; Second to study the role of expression of Thl/Th2 types cytokines & transcription factor T-bet mRNA in PBMC in the pathogenesis of alopecia areata and its regulation by Procyanidins(PC) and Astraglus polysaccharide(APS).Methods Apply RT-PCR technique to detect the mRNA expression of Thl/Th2 types cytokines & transcription factor T-bet in PBMC of the patients with alopecia areata by stimulation with phytohemagglutinin(PHA) and PHA plus PC/APS respectively.Results The mRNA expression of Thl type cytokines in PHA-stimulated PBMC of the patients with localized alopecia areata were increased slightly compared with healthy controls, the expression of transcription factor T-bet mRNA markedly elevated with the healthy controls (p<0. 05) , on the contrary, the mRNA expression of Th2 type cytokines (IL-10)lower significantly than healthy controls(P<0. 05). However, the mRNA expression of both Thltype cytokines & transcription factor T-bet in PHA-stimulated PBMC of the patients with extensive alopecia areata were markedly increased compared with healthy controls(P<0. 01)and localized form(P<0. 05), but the mRNA expression of Th2 type cytokines (IL-10)were lower significantly than healthy
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