摘要
目的比较HBV感染免疫耐受期产妇脐带血与正常产妇脐带血浆细胞样树突状细胞(pDC)中PI3K、mTOR、p70S6K、IFNα表达的差异。方法选取湖南中医药大学第一附属医院产科住院部2017年10月-2020年1月HBV感染免疫耐受期产妇20例(乙型肝炎组)及正常产妇10例(正常组)。分离培养脐带血pDC,在培养的第7天加入CpG-A,24 h后分别收集细胞及上清液,real-time PCR检测pDC PI3K、mTOR、p70S6K mRNA的表达,Western Blot技术检测pDC PI3K、mTOR、p70S6K蛋白表达,ELISA法检测pDC培养上清IFNα水平。计量资料采用两独立样本t检验。结果乙型肝炎组HBV感染免疫耐受期产妇脐带血pDC PI3K、mTOR、p70S6K mRNA表达水平显著低于正常产妇组(t值分别为-81.04、-63.07、-34.55,P值均<0.001);乙型肝炎组HBV感染免疫耐受期产妇脐带血pDC PI3K、mTOR、p70S6K蛋白表达水平亦显著低于正常产妇组(t值分别为-8.13、-7.75、-6.71,P值均<0.001);乙型肝炎组HBV感染免疫耐受期产妇脐带血pDC培养上清IFNα表达水平显著低于正常产妇组(t=-15.88,P<0.05)。结论HBV感染免疫耐受期产妇脐带血pDC的PI3K、mTOR、p70S6K mRNA和蛋白表达水平及IFNα水平减少,pDC功能受到抑制。
Objective To investigate the expression of phosphoinositide 3-kinase(PI3K),mammalian target of rapamycin(mTOR),70-kDa ribosomal protein S6 kinase(p70S6K),and interferon-α(IFNα)in umbilical cord blood plasmacytoid dendritic cells(pDCs)in parturients in the immune tolerance stage of hepatitis B virus(HBV)infection versus normal parturients.Methods A total of 20 parturients in the immune tolerance stage of HBV infection and 10 normal parturients who were hospitalized in Inpatient Department of Obstetrics in The First Affiliated Hospital of Hunan University of Chinese Medicine from October 2017 to January 2020 were enrolled as hepatitis B group and normal group,respectively.Umbilical cord blood pDCs were isolated and cultured,and CpG-A was added on day 7.The cells and the supernatant were collected after 24 hours;real-time PCR was used to measure the mRNA expression of PI3K,mTOR,and p70S6K,Western Blot was used to measure the protein expression of PI3K,mTOR,and p70S6K,and ELISA was used to measure the level of IFNαin the supernatant of pDCs.The two-independent-samples t-test was used for comparison of continuous data between the two groups.Results Compared with the normal group,the hepatitis B group had significantly lower mRNA and protein expression of PI3K,mTOR,and p70S6K in umbilical cord blood pDCs(mRNA expression:t=-81.04,-63.07,and-34.55,all P<0.001;protein expression:t=-8.13,-7.75,and-6.71,all P<0.001).The hepatitis B group had significantly lower expression of IFNαin the supernatant of umbilical cord blood pDCs than the normal group(t=-15.88,P<0.05).Conclusion Parturients in the immune tolerance stage of HBV infection have reductions in the mRNA and protein expression of PI3K,mTOR,and p70S6K and the level of IFNαin umbilical cord blood pDCs,suggesting that pDC function is inhibited.
作者
杨雯珺
霍燚
彭建平
YANG Wenjun;HUO Yi;PENG Jianping(Hunan University of Chinese Medicine,Changsha 410007,China;Department of Hepatology,The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,China)
出处
《临床肝胆病杂志》
CAS
北大核心
2020年第8期1727-1731,共5页
Journal of Clinical Hepatology
基金
国家自然科学基金项目(8170151973)。
