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目的探讨不明原因复发性流产与蜕膜组织中巨噬细胞自噬状态之间的关系。方法根据流产情况分为不明原因复发性流产组(观察组)和人工流产组(对照组),每组病例各30例。将蜕膜组织制成细胞悬液后分离淋巴细胞,通过CD14+免疫磁珠分选巨噬细胞,抗F4/80检测纯度,比较两组中巨噬细胞数量。通过免疫组化检测蜕膜中巨噬细胞表达,评价巨噬细胞与流产间关系;通过RT-q PCR检测细胞自噬相关基因Atg5,Atg7,LC3-Ⅱ和Bnip3表达水平的改变;Western Blot检测LC3-Ⅱ水平以评价自噬状态,对比两组之间差异。结果观察组中巨噬细胞纯度为(88.87±3.89)%,对照组为(82.84±3.98)%,差异有统计学意义(t=5.93,P<0.01)。巨噬细胞在观察组中阳性表达率为96.7%(29/30)高于对照组中的80.0%(24/30)。观察组中自噬相关基因ATG5、ATG7、LC3-Ⅱ和Bnip3的m RNA表达水平高于对照组,差异有统计学意义(t值分别为2.90,2.67,12.00,12.13,P值均<0.01)。Western Blot检测显示观察组中LC3-Ⅱ相对表达量为0.51±0.23,对照组为0.24±0.09,差异有统计学意义(t=5.96,P<0.01)。结论在不明原因复发性流产病例中,蜕膜中巨噬细胞的自噬高于对照组,巨噬细胞的自噬与不明原因复发性流产的发生有一定的相关性。
Objective To explore the relationship between unexplained recurrent spontaneous abortion and macrophage autophagy in decidua. Methods According to abortion, the patients were divided into unexplained recurrent spontaneous abortion group (observation group) and induced abortion group (control group), 30 cases in each group. After the decidual tissue was made into cell suspension, lymphocytes were separated, macrophages were sorted by CD14 + immunomagnetic beads, anti-F4 / 80 purity was tested, and the number of macrophages in both groups was compared. The expression of macrophages in macrophages and abortion was evaluated by immunohistochemistry. The expression of Atg5, Atg7, LC3-Ⅱ and Bnip3 in autophagy-related genes was detected by RT-q PCR. LC3-II levels to evaluate the autophagy status, comparing the differences between the two groups. Results The purity of macrophages in the observation group was (88.87 ± 3.89)% and that in the control group was (82.84 ± 3.98)%, the difference was statistically significant (t = 5.93, P <0.01). The positive rate of macrophages in the observation group was 96.7% (29/30), which was higher than that in the control group (80.0%, 24/30). The m RNA expression of autophagy-related genes ATG5, ATG7, LC3-Ⅱ and Bnip3 in the observation group was significantly higher than that in the control group (t = 2.90,2.67,12.00,12.13, P <0.01) . Western Blot showed that the relative expression level of LC3-Ⅱ was 0.51 ± 0.23 in the observation group and 0.24 ± 0.09 in the control group, the difference was statistically significant (t = 5.96, P <0.01). Conclusions Autophagy in macrophages in decidua is higher than that in control group in cases of unexplained recurrent spontaneous abortion. Autophagy in macrophages is correlated with the occurrence of unexplained recurrent spontaneous abortion.