【摘要】 目的 观察早发性卵巢功能不全(POI)患者外周血滤泡辅助性T细胞(Tfh)占比、功能相关细胞因子表达变化，并分析其与性激素水平的相关性。方法 POI患者29例(观察组)，同期健康体检者31例(对照组)，采用流式细胞术检测全血Tfh占比，免疫印迹实验检测外周血单个核细胞(PBMC)CXCR5、BCL-6蛋白，酶联免疫吸附法检测血清IL-6、IL-21，电化学发光法检测血清促卵泡刺激素(FSH)、促黄体生成素(LH)、泌乳素、孕酮、睾酮、雌二醇(E2)、抗缪勒管激素(AMH)水平，Spearman相关分析法分析Tfh、CXCR5蛋白、BCL-6蛋白、IL-6、IL-21与性激素的相关性。结果 与对照组比较，观察组全血Tfh占比增加，血清IL-6、IL-21水平升高，PBMC CXCR5、BCL-6蛋白表达升高，血清FSH、LH水平升高，血清E2、AMH水平降低，P均<0.05。全血Tfh占比与血清FSH、LH水平呈正相关(r分别为0.75、0.63,P均<0.05)，与血清E2、AMH水平呈负相关(r分别为-0.35、-0.82,P均<0.05)；血清IL-6水平与血清FSH、LH水平呈正相关(r分别为0.75、0.62,P均<0.05)，与血清E2、AMH水平呈负相关(r分别为-0.47、-0.77,P均<0.05)；血清IL-12水平与血清FSH、LH水平呈正相关(r分别为0.68、0.56,P均<0.05)，与血清E2、AMH水平呈负相关(r分别为-0.25、-0.73,P均<0.05);PBMC BCL-6蛋白表达与血清FSH、LH水平呈正相关(r分别为0.73、0.60,P均<0.05)，与血清E2、AMH水平呈负相关(r分别为-0.27、-0.76,P均<0.05);PBMC CXCR5蛋白表达与血清FSH、LH水平呈正相关(r分别为0.72、0.58,P均<0.05)，与血清E2、AMH水平呈负相关(r分别为-0.27、-0.70,P均<0.05)。结论 POI患者全血Tfh占比增加，PBMC CXCR5、BCL-6蛋白表达升高，血清IL-6、IL-21、FSH、LH水平升高及E2、AMH水平降低，Tfh及其相关因子与性激素呈正相关或负相关，Tfh及其相关因子可能参与了POI发生过程。更多还原

【Abstract】 Objective To observe the changes in the proportion of peripheral blood T follicular helper(Tfh) cells and the expression of function-related cytokines in patients with premature ovarian insufficiency(POI),and to analyze their correlations with sex hormone levels.Methods We recruited 29 POI patients(observation group) and 31 healthy individuals(control group) for physical examination during the same period.We used flow cytometry to detect the proportion of Tfh in whole blood,immunoblotting to measure the expression of CXCR5 and BCL-6 proteins in peripheral blood mononuclear cells(PBMCs),and ELISA to detect serum levels of IL-6 and IL-21.Electrochemiluminescence immunoassay was employed to determine serum levels of follicle-stimulating hormone(FSH),luteinizing hormone(LH),prolactin,progesterone,testosterone,estradiol(E2),and anti-Mullerian hormone(AMH).Spearman’s correlation analysis was performed to assess the correlations between Tfh,CXCR5 protein,BCL-6 protein,IL-6,IL-21,and sex hormones.Results Compared with the control group,the observation group exhibited an increased proportion of Tfh in whole blood,elevated serum levels of IL-6 and IL-21,up-regulated expression of CXCR5 and BCL-6 proteins in PBMCs,higher serum levels of FSH and LH,and lower serum levels of E2 and AMH,with statistically significant differences(all P<0.05).The proportion of Tfh in whole blood was positively correlated with serum levels of FSH and LH(r =0.75 and 0.63,respectively; both P<0.05),and was negatively correlated with serum levels of E2 and AMH(r=–0.35 and-0.82,respectively; both P<0.05).Similarly,serum IL-6 level was positively correlated with serum FSH and LH levels(r=0.75 and 0.62,respectively; both P < 0.05) and was negatively correlated with serum E2 and AMH levels(r=–0.47 and-0.77,respectively; both P<0.05).Additionally,serum IL-12 level was positively correlated with serum FSH and LH levels(r=0.68 and 0.56,respectively; both P<0.05) and was negatively correlated with serum E2 and AMH levels(r=–0.25 and–0.73,respectively; both P<0.05).Expression of BCL-6 protein in PBMCs was positively correlated with serum FSH and LH levels(r=0.73 and 0.60,respectively; both P<0.05) and was negatively correlated with serum E2 and AMH levels(r=–0.27 and–0.76,respectively; both P<0.05).Similarly,the expression of CXCR5 protein in PBMCs was positively correlated with serum FSH and LH levels(r=0.72 and 0.58,respectively; both P<0.05) and was negatively correlated with serum E2 and AMH levels(r=–0.27 and–0.70,respectively; both P<0.05).Conclusions In POI patients,there is an increased proportion of Tfh in whole blood,up-regulated expression of CXCR5 and BCL-6 proteins in PBMCs,elevated serum levels of IL-6,IL-21,FSH,and LH,as well as decreased levels of E2 and AMH.Tfh and its related factors show positive or negative correlations with sex hormones,suggesting that Tfh and its associated factors may be involved in the pathogenesis of POI.更多还原