外周血TBNK淋巴细胞亚群和血清Th1/Th2细胞因子与不明原因复发性流产的相关性研究

Study on correlation between peripheral blood TBNK lymphocyte subsets and serum Th1/Th2 cytokines with unexplained recurrent spontaneous abortion

目的:探讨外周血TBNK淋巴细胞亚群和血清Th1/Th2细胞因子与不明原因复发性流产(URSA)的相关性。方法:淋巴细胞亚群和Th1/Th2细胞因子均采用流式细胞术检测;选取2018年5月至2019年5月成都西囡妇科医院就诊的60例不明原因复发性流产女性患者为URSA病例组,同时选择年龄匹配的近1年有健康分娩史的30例妇女为正常对照组。采用流式细胞术分别检测两组人群外周血TBNK淋巴细胞亚群(CD3^(+),CD4^(+),CD8^(+),CD19^(+)和CD16^(+)CD56^(+))的比例和绝对值计数及Th1/Th2细胞因子(IL-2,TNF-α,IFN-γ/IL-4,IL-6和IL-10)含量,并对检测结果进行统计学分析。对其中40例URSA患者进行不同妊娠结局随访,对22例正常妊娠与18例再次流产患者外周血淋巴亚群和Th1/Th2细胞因子的结果进行比较分析。回顾性分析其中22例URSA患者正常妊娠前后外周血TBNK淋巴亚群与Th1/Th2细胞因子的变化。结果:①URSA病例组外周血的Th细胞(CD4^(+)及CD4/^(+)CD8^(+)),B细胞(CD19^(+))以及NK细胞(CD16^(+)CD56^(+))比例和绝对值计数结果均高于正常对照组,差异有统计学意义(P<0.05)。②URSA病例组外周血中TNF-α、IFN-γ和IL-6均高于正常对照组,差异有统计学意义(P<0.05)。③再次流产URSA患者外周血的Th细胞,B细胞以及NK细胞比例和绝对值计数结果均高于正常妊娠组,差异有统计学意义(P<0.05),同时再次流产URSA患者外周血中的TNF-α、IFN-γ和IL-6均高于正常妊娠组,差异有统计学意义(P<0.05)。④正常妊娠URSA患者临床治疗前后作为自身对比,结果显示治疗后和正常妊娠后外周血Th细胞,B细胞以及NK细胞比例和绝对值计数结果均比妊娠前有明显下降,差异有统计学意义(P<0.05)。同时,治疗后与妊娠后外周血中TNF-α、IFN-γ和IL-6均比妊娠前有所下降,差异有统计学意义(P<0.05)。结论:URSA患者体内免疫功能发生了紊乱,患者外周血的淋巴细胞亚群比例和数量的异常以及体内Th1/Th2细胞因子的失衡可能是导致不明原因流产的重要因素,其发病机制可能与体内Th细胞、B细胞和NK细胞增高有关,也可能同时与TNF-α和IFN-γ(Th1型细胞因子)或者IL-6(Th2型细胞因子)异常增高有关。URSA患者外周血TBNK淋巴细胞与Th1/Th2细胞细胞的变化与妊娠结局相关。

Objective:To investigate the correlation between TBNK lymphocyte subsets and Th1/Th2 cytokines in peripheral blood with unexplained recurrent abortion(URSA).Methods:lymphocyte subsets and Th1/Th2 cytokines were detected by flow cytometry.60 URSA women who came to Xi′nan Gynecological Hospital from May 2018 to May 2019 were recruited as the URSA case group,meanwhile,age matched 30 healthy women with a history of healthy delivery in the past one year were selected as the healthy control group.The ratio and absolute value of TBNK lymphocyte subsets(CD3^(+),CD4^(+),CD8^(+),CD19^(+)and CD16^(+)CD56^(+))and Th1/Th2 cytokines(IL-2,TNF-α,IFN-γ/IL-4,IL-6 and IL-10)were measured by flow cytometry,and the results were analyzed statisti⁃cally.40 URSA patients were followed up and divide into two groups according to the pregnancy outcomes.The results of peripheral blood lymphoid subsets and Th1/Th2 cytokines in 22 URSA patients with normal pregnancy and 18 URSA with recurrent abortion were compared and analyzed.The changes of TBNK lymphoid subsets and Th1/Th2 cytokines in peripheral blood of 22 URSA patients before and after normal pregnancy were analyzed retrospectively.Results:①The percentage and absolute value of Th cells(CD4^(+)and CD4/^(+)CD8^(+)),B cells(CD19^(+))and NK cells(CD16^(+)CD56^(+))in the peripheral blood of the URSA case group were higher than those of the healthy control group(P<0.05).②TNF-α,IFN-γand IL-6 in the URSA case group was significantly higher than that of the healthy control group(P<0.05).③The percentage and absolute value of Th cells,B cells and NK cells in the peripheral blood of URSA patients with recurrent abortion were significantly higher than those of URSA patients with normal pregnancy(P<0.05),Meanwhile,TNF-α,IFN-γand IL-6 in URSA patients with recurrent abortion were higher than those of URSA patients with normal pregnancy group,the difference was statistically significant(P<0.05).④The results showed that the percentage and absolute value of Th cells,B cells and NK cells in the peripheral blood of URSA patients with normal pregnancy before and after treatment were significantly lower than those before treatment(P<0.05).At the same time,TNF-α,IFN-γand IL-6 after treatment and after pregnancy were lower than those before pregnancy(P<0.05).Conclusion:The disorder of immune function in URSA patients may be caused by the abnormal proportion and number of lymphocyte subsets in peripheral blood and the imbalance of Th1/Th2 cytokines.The pathogenesis of URSA may be related to the increase of Th cells,B cells and NK cells,as well as the increase of TNF-αand IFN-γ(Th1 type cytokine)or IL-6(Th2 type cytokine)increased abnormally.The changes of TBNK cells and Th1/Th2 cells in URSA patients were correlated with pregnancy outcome.