二甲双胍对复发性流产大鼠妊娠结局、内分泌及VEGF/MAPK信号通路的影响

doi:10.11958/20221259

天津医药 ›› 2023, Vol. 51 ›› Issue (5): 482-486.doi: 10.11958/20221259

二甲双胍对复发性流产大鼠妊娠结局、内分泌及VEGF/MAPK信号通路的影响

周知(), 黎业娟, 陈琳, 王安国, 阮海玲, 马宁, 周璟, 卢伟英^△()

海南省妇女儿童医学中心，生殖医学中心（邮编570206）

收稿日期:2022-08-15 修回日期:2022-12-02 出版日期:2023-05-15 发布日期:2023-05-05
通讯作者: △E-mail:2490206511@qq.com
作者简介:周知（1981），女，副主任医师，主要从事生殖内分泌、医学遗传学方面研究。E-mail：ztd681@163.com
基金资助:
海南省重点研发计划项目(ZDYF2017086);海南省重点研发计划项目(ZDYF2019157);海南省基础与应用基础研究计划（自然科学领域）高层次人才项目(2019RCJ89)

Effects of metformin on pregnancy outcome, endocrine and VEGF/MAPK signaling pathway in rats with recurrent spontaneous abortion

ZHOU Zhi(), LI Yejuan, CHEN Lin, WANG Anguo, RUAN Hailing, MA Ning, ZHOU Jing, LU Weiying^△()

Reproductive Medicine Center, Hainan Women and Children’s Medical Center, Haikou 570206, China

Received:2022-08-15 Revised:2022-12-02 Published:2023-05-15 Online:2023-05-05
Contact: △E-mail:2490206511@qq.com

周知, 黎业娟, 陈琳, 王安国, 阮海玲, 马宁, 周璟, 卢伟英. 二甲双胍对复发性流产大鼠妊娠结局、内分泌及VEGF/MAPK信号通路的影响[J]. 天津医药, 2023, 51(5): 482-486.

ZHOU Zhi, LI Yejuan, CHEN Lin, WANG Anguo, RUAN Hailing, MA Ning, ZHOU Jing, LU Weiying. Effects of metformin on pregnancy outcome, endocrine and VEGF/MAPK signaling pathway in rats with recurrent spontaneous abortion[J]. Tianjin Medical Journal, 2023, 51(5): 482-486.

摘要/Abstract

摘要：

目的探讨二甲双胍（Met）对复发性流产（RSA）大鼠血管内皮生长因子（VEGF）/丝裂原活化蛋白激酶（MAPK）信号通路及卵巢内分泌的影响。方法 SD雌性大鼠与雄性大鼠按照2∶1的比例合笼交配后将受孕成功的雌性大鼠采用随机数字表法分为对照组，RSA组，Met低、中、高剂量组（250、1 000、2 000 mg/kg），黄体酮组（阳性对照组，156 mg/kg），每组10只。末次给药结束后24 h，将大鼠麻醉后处死，解剖大鼠子宫观察胚胎丢失情况；苏木精-伊红染色检测子宫蜕膜组织病理变化；放射免疫试剂盒检测血清激素水平；蛋白免疫印迹法检测子宫蜕膜组织中VEGF/MAPK通路蛋白水平。结果与对照组比较，RSA组大鼠死亡胚胎数及胚胎丢失率升高，血清卵泡刺激素（FSH）、黄体生成激素（LH）、垂体泌乳素（PRL）、雌二醇（E₂）、孕酮（P）表达量以及子宫蜕膜组织中VEGF受体-2（VEGFR-2）、VEGF、磷酸化细胞外信号调节激酶（p-ERK）蛋白表达量均降低（P<0.05），子宫蜕膜组织受损严重；与RSA组比较，Met各剂量组大鼠死亡胚胎数及胚胎丢失率降低，血清FSH、LH、PRL、E₂及P表达量以及子宫蜕膜组织中VEGFR-2、VEGF、p-ERK蛋白表达量均升高（P<0.05），子宫蜕膜组织损伤得到一定的缓解，且高剂量对上述指标的控制效果更好。结论 Met可缓解RSA大鼠内分泌失调及子宫蜕膜损伤，提高VEGF/MAPK通路相关蛋白的表达，具有促进VEGF/MAPK通路激活的作用。

关键词: 二甲双胍, 流产,习惯性, 内皮生长因子类, 丝裂原激活蛋白激酶类

Abstract:

Objective To investigate effects of metformin (Met) on vascular endothelial growth factor (VEGF)/mitogen-activated protein kinase (MAPK) signaling pathway and ovarian endocrine in recurrent abortion (RSA) rats. Methods Female SD rats were mated with male rats in a ratio of 2∶1, then the female pregnant rats were randomly divided into the control group, the RSA group, the Met low-dose, middle-dose and high-dose groups (250, 1 000, 2 000 mg/kg) and the progesterone group (positive control group, 156 mg/kg), with 10 rats in each group. At 24 hours after the last administration, rats were anesthetized and killed. The embryo loss was observed by dissecting rat uterus. Hematoxylin eosin (HE) staining was used to detect pathological changes of decidual tissue. Radioimmunoassay kit was used to detect serum hormone levels. The protein levels of VEGF/MAPK pathway in uterine decidua were detected by Western blot assay. Results Compared with the control group, the number of dead embryos and the rate of embryo loss were significantly increased in the RSA group, and serum levels of follicle-stimulating hormone (FSH), leuteinizing hormone (LH), pituitary prolactin (PRL), estradiol (E₂), progesterone (P) and protein expressions of VEGF receptor-2 (VEGFR-2), VEGF and phosphorylated extracellular signal-regulated kinase (p-ERK) in decidual tissue were significantly decreased (P<0.05), and the decidual tissue of uterus was seriously damaged. Compared with the RSA group, the number of dead embryos and the rate of embryo loss were significantly decreased in the Met groups. Serum levels of FSH, LH, PRL, E₂, P and the protein expression levels of VEGFR-2, VEGF and p-ERK in decidual tissue were significantly increased (P<0.05). The damage of decidual tissue was alleviated, and high dose of Met showed better control effect on the above indexes. Conclusion Met can alleviate endocrine disorder and uterine decidua damage in RSA rats, which significantly improves the expression of VEGF/MAPK pathway related proteins, and promotes the activation of VEGF/MAPK pathway.

Key words: metformin, abortion, habitual, endothelial growth factors, mitogen-activated protein kinases

中图分类号:

R730.21

图/表 6

参考文献 21

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组别	存活胚胎数（只）	死亡胚胎数（只）	胚胎丢失率（%）
对照组	56	5	8.19±1.70
RSA组	25	30	54.54±8.29^a
Met低剂量组	31	26	45.61±7.44^b
Met中剂量组	42	19	31.14±7.30^bc
Met高剂量组	49	7	12.50±2.52^bcd
黄体酮组	50	10	16.66±3.36^bcd
F			108.271^＊＊

组别	存活胚胎数（只）	死亡胚胎数（只）	胚胎丢失率（%）
对照组	56	5	8.19±1.70
RSA组	25	30	54.54±8.29^a
Met低剂量组	31	26	45.61±7.44^b
Met中剂量组	42	19	31.14±7.30^bc
Met高剂量组	49	7	12.50±2.52^bcd
黄体酮组	50	10	16.66±3.36^bcd
F			108.271^＊＊

组别		FSH（IU/L）		LH（IU/L）
对照组		7.01±0.27		6.03±0.75
RSA组		5.07±0.11^a		3.09±0.39^a
Met低剂量组		5.71±0.13^b		3.92±0.49^b
Met中剂量组		6.20±0.17^bc		4.86±0.60^bc
Met高剂量组		6.84±0.25^bcd		5.83±0.72^bcd
黄体酮组		6.72±0.24^bcd		5.61±0.70^bcd
F		135.481^＊＊		35.556^＊＊
组别	PRL（mIU/L）		E₂（ng/L）		P（μg/L）
对照组	102.61±5.40		24.20±0.20		2.40±0.21
RSA组	70.06±2.08^a		17.11±0.45^a		1.14±0.10^a
Met低剂量组	78.19±3.01^b		18.71±0.12^b		1.57±0.14^b
Met中剂量组	86.37±3.71^bc		19.94±0.42^bc		1.90±0.17^bc
Met高剂量组	96.37±4.26^bcd		22.46±0.81^bcd		2.28±0.20^bcd
黄体酮组	95.13±4.31^bcd		21.97±0.87^bcd		2.19±0.19^bcd
F	97.383^＊＊		223.454^＊＊		78.160^＊＊

组别		FSH（IU/L）		LH（IU/L）
对照组		7.01±0.27		6.03±0.75
RSA组		5.07±0.11^a		3.09±0.39^a
Met低剂量组		5.71±0.13^b		3.92±0.49^b
Met中剂量组		6.20±0.17^bc		4.86±0.60^bc
Met高剂量组		6.84±0.25^bcd		5.83±0.72^bcd
黄体酮组		6.72±0.24^bcd		5.61±0.70^bcd
F		135.481^＊＊		35.556^＊＊
组别	PRL（mIU/L）		E₂（ng/L）		P（μg/L）
对照组	102.61±5.40		24.20±0.20		2.40±0.21
RSA组	70.06±2.08^a		17.11±0.45^a		1.14±0.10^a
Met低剂量组	78.19±3.01^b		18.71±0.12^b		1.57±0.14^b
Met中剂量组	86.37±3.71^bc		19.94±0.42^bc		1.90±0.17^bc
Met高剂量组	96.37±4.26^bcd		22.46±0.81^bcd		2.28±0.20^bcd
黄体酮组	95.13±4.31^bcd		21.97±0.87^bcd		2.19±0.19^bcd
F	97.383^＊＊		223.454^＊＊		78.160^＊＊

组别	VEGFR-2/GAPDH	VEGF/GAPDH	p-ERK/ERK
对照组	1.16±0.23	1.14±0.22	1.18±0.04
RSA组	0.21±0.08^a	0.15±0.08^a	0.40±0.02^a
Met低剂量组	0.43±0.11^b	0.45±0.10^b	0.58±0.04^b
Met中剂量组	0.71±0.14^bc	0.74±0.15^bc	0.71±0.07^bc
Met高剂量组	0.94±0.18^bcd	0.96±0.19^bcd	1.05±0.09^bcd
黄体酮组	0.91±0.17^bcd	0.93±0.16^bcd	0.97±0.13^bcd
F	49.019^＊＊	54.434^＊＊	161.534^＊＊

二甲双胍对复发性流产大鼠妊娠结局、内分泌及VEGF/MAPK信号通路的影响

Effects of metformin on pregnancy outcome, endocrine and VEGF/MAPK signaling pathway in rats with recurrent spontaneous abortion

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