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关系探讨  

Study of the relationship between unexplained recurrent spontaneous abortion and macrophage autophagy in decidua tissue

作　　者：杜涛[1] 黄海[1] 刘玉昆[1] 张睿[1] 丁红[1] 陈欣[1] 张建平[1] 

机构地区：[1]中山大学孙逸仙纪念医院妇产科,广东广州510120

出　　处：《中国实用妇科与产科杂志》2015年第8期763-767,共5页Chinese Journal of Practical Gynecology and Obstetrics

基　　金：广东省科技社会发展项目(2012B032000006)

摘　　要：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 study the relationship between unexplained recurrent spontaneous abortion and decidua mac- rophage autophagy. Methods According to the cause of abortion, the patients were divided into unexplained recurrent spontaneous abortion groups and artificially abortion groups with 30 patients in each group. The acquired decidua was made into cell suspension, and lymphocytes isolated. Macrophages were dissociated through immunomagnetic by CD 14＋ cells, and antibody of F4/80 was used to test purity of macrophage. The number of macrophage was compared between two groups. The comparison of decidual maerophages expression of two groups was made by immunohistochemistry. RT- qPCR was used to detect autophagy genes Atg5, Atg7, LC3- Ⅱ and Bnip3 expression levels, and Western Blot was used to detect LC3- Ⅱ levels in order to evaluate the state of autophagy between the two groups.Results The maerophage purity in unexplained recurrent spontaneous abortion group was （88.87 ± 3.89）%, and （82.84 ± 3.98）% in artificially abortion group. There was significant difference between the two groups. Macrophages were expressed in two groups, but in cases of unexplained recurrent spontaneous abortion the positive rate was 96.7% （29/30）, higher than the 80.0% （24/30） in the artificially abortion group. The mRNA expression levels of autophagy-related genes ATG5, ATG7, LC3- Ⅱ and Bnip3 were statistically different between two groups, and the unexplained recurrent spontaneous abortion group was higher than artificially abortion group. Western Blot analysis showed the LC3- Ⅱ expression was 0.51±0.23 in unexplained recurrent spontaneous abortion group,and compared to the artificially group 0.24 ± 0.09,the significant differences existed between the two groups. Conclusions In cases of unexplained recurrent spontaneous abortion, the macrophage autophagy in decidua is higher than control group, and the autophagy state of macrophage has certain relevance with unexplained recurrent spontaneous abortion.

关 键 词：复发性自然流产 自噬 巨噬细胞 蜕膜 

分 类 号：R714.21[医药卫生—妇产科学]