IVF/ICSI助孕后早期流产患者蜕膜组织焦亡相关因子表达水平的研究

doi:10.11958/20220918

天津医药 ›› 2023, Vol. 51 ›› Issue (4): 351-354.doi: 10.11958/20220918

IVF/ICSI助孕后早期流产患者蜕膜组织焦亡相关因子表达水平的研究

车小群(), 王丽莲, 王笑, 许丽华, 马天仲, 荆霞, 陈碧()

广东医科大学附属医院生殖中心（邮编524001）

收稿日期:2022-06-21 修回日期:2022-09-28 出版日期:2023-04-15 发布日期:2023-04-20
通讯作者: 陈碧 E-mail:xiaoqunche@126.com;chenbi9677@163.com
作者简介:车小群（1977），女，副主任医师，主要从事辅助生殖早期流产方面研究。E-mail：xiaoqunche@126.com
基金资助:
2017年度广东省医学科研基金立项项目(A2017596)

Study on the expression of pyrolysis-related factors in the decidua tissue of patients with early abortion after IVF/ICSI assisted pregnancy

CHE Xiaoqun(), WANG Lilian, WANG Xiao, XU Lihua, MA Tianzhong, JING Xia, CHEN Bi()

Reproductive Center, Affiliated Hospital of Guangdong Medical University, Zhanjiang 524001, China

Received:2022-06-21 Revised:2022-09-28 Published:2023-04-15 Online:2023-04-20
Contact: CHEN Bi E-mail:xiaoqunche@126.com;chenbi9677@163.com

车小群, 王丽莲, 王笑, 许丽华, 马天仲, 荆霞, 陈碧. IVF/ICSI助孕后早期流产患者蜕膜组织焦亡相关因子表达水平的研究[J]. 天津医药, 2023, 51(4): 351-354.

CHE Xiaoqun, WANG Lilian, WANG Xiao, XU Lihua, MA Tianzhong, JING Xia, CHEN Bi. Study on the expression of pyrolysis-related factors in the decidua tissue of patients with early abortion after IVF/ICSI assisted pregnancy[J]. Tianjin Medical Journal, 2023, 51(4): 351-354.

摘要/Abstract

摘要：

目的探讨体外受精（IVF）/卵胞浆内单精子注射（ICSI）助孕妊娠后早期流产患者蜕膜组织中焦亡相关因子的表达及作用。方法选取行IVF/ICSI助孕妊娠后早期流产患者102例（研究组），同期因意外妊娠行人工流产术的患者100例（对照组），收集蜕膜组织。实时荧光定量PCR（qRT-PCR）法及Western blot法检测蜕膜组织中焦亡相关因子Nod样受体蛋白3（NLRP3）、白细胞介素（IL）-1β、胱天蛋白酶1（Caspase1）、gasdermin D（GSDMD） mRNA及蛋白表达，TUNEL法检测蜕膜组织中细胞焦亡情况，Pearson法分析蜕膜组织中细胞焦亡率与NLRP3、IL-1β、Caspase1、GSDMD蛋白水平的相关性。结果与对照组相比，研究组蜕膜组织中NLRP3、IL-1β、Caspase1、GSDMD mRNA及蛋白水平、蜕膜组织中细胞焦亡率升高（P<0.05）；研究组患者蜕膜组织中细胞焦亡率与NLRP3、IL-1β、Caspase1、GSDMD蛋白表达水平均呈正相关（r分别为0.429、0.474、0.498、0.540，均P<0.05）。结论 IVF/ICSI助孕妊娠后早期流产患者蜕膜组织中焦亡相关因子NLRP3、IL-1β、Caspase1、GSDMD表达均升高，且与蜕膜组织细胞焦亡有关。

关键词: 流产，治疗性, 受精，体外, 精子注射，细胞质内, 蜕膜组织, Nod样受体蛋白3, 白细胞介素-1β, 胱天蛋白酶1, gasdermin D

Abstract:

Objective To investigate the expression and effect of pyrodeath related factors in decidual tissue of in vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) assisted early postpregnancy abortion. Methods A total of 102 patients with early abortion after IVF/ICSI assisted pregnancy were selected as the research group, and 100 patients who underwent artificial abortion due to unexpected pregnancy during the same period were selected as the control group. Samples of decidua tissue were collected. Quantitative real-time PCR (qRT-PCR) method and Western blot assay were used to detect expression levels of Nod-like receptor pyrin domain 3 (NLRP3), interleukin-1β (IL-1β), Cysteinyl aspartate specific proteinase 1 (Caspase1), gasdermin D (GSDMD) mRNA and protein in decidua tissue. TUNEL method was used to detect the cell pyroptosis in decidua tissue. Pearson method was used to analyze the correlation between pyroptosis rate and the protein levels of NLRP3, IL-1β, Caspase1 and GSDMD in decidua. Results Compared with the control group, expression levels of NLRP3, IL-1β, Caspase1, GSDMD mRNA and protein, the cell pyroptosis rate in decidua tissues were significantly increased in the research group (P<0.05). There was a positive correlation between cell pyroptosis rate in decidua tissue and expression levels of NLRP3, IL-1β, Caspase1 and GSDMD in the research group (r=0.429, 0.474, 0.498, 0.540, all P<0.05). Conclusion The expression levels of pyrolysis-related factors NLRP3, IL-1β, Caspase1 and GSDMD in decidua tissue are increased in patients with early abortion after IVF/ICSI assisted pregnancy, and which are related to the cell pyroptosis of decidua tissue.

Key words: abortion, therapeutic, fertilization in vitro, sperm injections, intracytoplasmic, decidua tissues, Nod-like receptor pyrin domain 3, interleukin-1β, cysteinyl aspartate specific proteinase 1, gasdermin D

中图分类号:

R711.6

图/表 5

表1 引物序列

Tab.1 primer sequence

基因名称	引物序列（5'→3'）	产物大小（bp）
NLRP3	上游：TGCCCGTCTGGGTGAGA	220
NLRP3	下游：CCGGTGCTCCTTGATGAGA	220
IL-1β	上游：CTCGCTTGAGAGGTGCTGATGGTAC	519
IL-1β	下游：GAAGCTGTGGCAGCTACCTATGTCT	519
Caspase1	上游：ACAAGGCACGGGACCTATG	237
Caspase1	下游：TCCCAGTCAGTCCTGGAAATG	237
GSDMD	上游：AGACAATAGACCCCTCCCC	362
GSDMD	下游：TCTGCTGCCGCTTACCTCC	362
GAPDH	上游：GGAGCGAGATCCCTCCAAAAT	197
GAPDH	下游：GGCTGTTGTCATACTTCTCATGG	197

表2 2组NLRP3、IL-1β、Caspase1、GSDMD mRNA表达水平比较（$\bar{x}±s$）

Tab.2 Comparison of the expression levels of NLRP3, IL-1β, Caspase1 and GSDMD mRNA in decidual tissue between the two groups of patients

组别	n	NLRP3	IL-1β	Caspase1	GSDMD
对照组	100	1.02±0.04	1.01±0.03	0.99±0.03	1.00±0.04
研究组	102	1.64±0.12	1.82±0.11	1.75±0.14	2.18±0.14
t		49.062^＊＊	71.087^＊＊	53.101^＊＊	81.099^＊＊

图1 2组患者蜕膜组织中NLRP3、IL-1β、Caspase1、GSDMD蛋白表达印迹图

Fig.1 Western blot results of NLRP3, IL-1β, Caspase1 and GSDMD protein expression in decidual tissue of the two groups of patients

表3 2组患者蜕膜组织中NLRP3、IL-1β、Caspase1、GSDMD蛋白表达水平比较（$\bar{x}±s$）

Tab.3 Comparison of protein expression levels of NLRP3, IL-1β, Caspase1 and GSDMD in decidual tissue between the two groups of patients

组别	n	NLRP3	IL-1β	Caspase1	GSDMD
对照组	100	0.24±0.04	0.28±0.05	0.30±0.04	0.22±0.04
研究组	102	0.44±0.08	0.55±0.12	0.50±0.11	0.62±0.14
t		22.404^＊＊	20.799^＊＊	17.106^＊＊	27.491^＊＊

图2 2组患者蜕膜组织细胞的TUNEL染色情况（TUNEL染色，×400）

Fig. 2 TUNEL staining of decidual tissue cells in the two groups of patients (TUNEL staining, ×400)

参考文献 17

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IVF/ICSI助孕后早期流产患者蜕膜组织焦亡相关因子表达水平的研究

Study on the expression of pyrolysis-related factors in the decidua tissue of patients with early abortion after IVF/ICSI assisted pregnancy

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