PI3K和Notch信号途径对寻常型银屑病患者外周血CD4^(+)T细胞的增殖和活化发挥协同调控作用  

PI3K and Notch signaling pathway play a synergistic regulatory role in the proliferation and activation of CD4^(+)T cells in peripheral blood of patients with psoriasis vulgaris

作　　者：刘佳[1] 魏明[1] LIU Jia;WEI Ming(Department of Clinical Laboratory,Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第五附属医院检验科,河南郑州450052

出　　处：《皮肤性病诊疗学杂志》2022年第4期305-310,共6页Journal of Diagnosis and Therapy on Dermato-venereology

摘　　要：目的研究PI3K和Notch信号途径在寻常型银屑病患者外周静脉血CD4^(+)T细胞增殖、活化中的影响。方法采集28例寻常型银屑病患者和28例健康对照外周静脉血,采用免疫磁珠法分选外周血CD4^(+)T细胞。将寻常型银屑病患者外周血CD4^(+)T细胞分为4组:空白对照组,10μmol/L PI3K阻滞剂组(LY294002组),25μmol/L Notch阻滞剂组(DAPT组),10μmol/L LY294002+25μmol/L DAPT组(LY294002+DAPT组)。均分别用植物血凝素(PHA)10μg/mL和IL-21000 U/mL刺激增殖6 h,采用FCM检测CD4^(+)T细胞内的CyclinA、Cyclin D1及P27^(kipl)细胞周期蛋白水平,逆转录-聚合酶链反应(RT-PCR)检测Cyclin A、Cyclin D1及P27^(kipl)mRNA水平,并对数据进行统计学处理。结果CD4^(+)T细胞经免疫磁珠法分选后纯度为(90.00±3.90)%,CD4^(+)T细胞培养存活百分率达(92.50±4.60)%。寻常型银屑病患者外周血CD4^(+)T细胞Cyclin A、Cyclin D1水平分别为(27.60±3.80)%、(14.70±3.20)%,mRNA水平分别为0.56±0.14、1.37±0.39,与健康对照组(13.50±3.70)%、(7.80±2.00)%和(0.31±0.12)、(0.93±0.35)比较均明显升高,差异有统计学意义(P分别为0.002、0.037和0.008、0.043)。寻常型银屑病患者外周血CD4^(+)T细胞P27^(kipl)蛋白和mRNA分别为(23.50±3.60)%和(0.17±0.02),均低于健康对照组的(36.80±5.60)%和(0.33±0.05),差异有统计学意义(P分别为0.007、0.001)。与空白对照组CD4^(+)T细胞Cyclin D1蛋白和mRNA表达[分别为(12.30±3.50)%和(1.74±0.39)]比较,LY294002组、DAPT组、LY294002+DAPT组表达均下降,分别为(7.20±3.10)%、(8.20±2.50)%、(4.30±1.50)%和(1.11±0.29)、(1.26±0.28)、(0.63±0.20),差异均有统计学意义(均P<0.05)。与空白对照组CD4^(+)T细胞P27^(kipl)蛋白[(21.80±3.20)%]比较,LY294002组、DAPT组、LY294002+DAPT组蛋白水平均升高,分别为(30.90±5.10)%、(27.60±5.20)%、(43.90±3.00)%,差异均有统计学意义(P分别为0.005、0.006、0.001);与空白对照组CD4^(+)T细胞P27^(kipl)mRNA(0.15±0.08)比较,LY294002�Objective To study effects of phosphoinositide 3 kinase(PI3K)and Notch signaling pathway on the proliferation and activation of CD4^(+)T cells in peripheral blood of patients with psoriasis vulgaris.Methods Peripheral venous blood was collected from 28 patients with psoriasis vulgaris and 28 healthy controls.CD4^(+)T cells in peripheral blood were sorted by immunomagnetic bead method.CD4^(+)T cells from peripheral blood of patients with psoriasis vulgaris were divided into four groups:the blank control group,and 10μmol/L PI3K blocker group(LY294002 group),25μmol/L Notch blocker group(DAPT group),10μmol/L LY294002+25μmol/L DAPT group(LY294002+DAPT group).CD4^(+)T cells were stimulated with phytohemagglutinin(PHA)10μg/ml and interleukin(IL)-21000 U/ml for 6 hours.The protein levels of Cyclina,Cyclin D1 and P27^(kipl)in CD4+T cells were detected by FCM,and the mRNA levels of Cyclin A,Cyclin D1 and P27^(kipl)were detected by reverse transcription polymerase chain reaction(RT-PCR).Results The purity of CD4^(+)T cells were(90.00±3.90)%,and the survival rate of CD4^(+)T cells was(92.50±4.60)%.The protein levels of Cyclin A and Cyclin D1 in peripheral blood CD4^(+)T cells of patients with psoriasis vulgaris were(27.60±3.80)%,(14.70±3.20)%,and the mRNA levels were(0.56±0.14)and(1.37±0.39)%,respectively.Compared with the healthy control group(13.50±3.70)%,(7.80±2.00)%and(0.31±0.12),(0.93±0.35),the difference was statistically significant(P=0.002,0.037,0.008 and 0.043).The level of P27^(kipl)Protein and mRNA of CD4^(+)T cells in peripheral blood of patients with psoriasis vulgaris were(23.50±3.60)%and(0.17±0.02)%respectively,which were significantly lower than those in healthy control group(36.80±5.60)%and(0.33±0.05)(P=0.007,P=0.001).The expression of Cyclin D1 protein and mRNA in CD4^(+)T cells of the blank control group were(12.30±3.50)%and(1.74±0.39),respectively,while the level in LY294002 group,DAPT group and LY294002+DAPT group decreased significantly,with(7.20±3.10)%,(8.20±2.50)%,(4.30±1.50)%a

关 键 词：寻常型银屑病 CD4^(+)T细胞 PI3K NOTCH 

分 类 号：R758.63[医药卫生—皮肤病学与性病学]