养血安胎方调节自噬治疗复发性自然流产小鼠的机制研究

doi:10.19803/j.1672-8629.20240060

中国药物警戒 ›› 2024, Vol. 21 ›› Issue (4): 378-385.
DOI: 10.19803/j.1672-8629.20240060

• 养血安胎方对复发性自然流产的作用机制研究专栏 • 上一篇下一篇

养血安胎方调节自噬治疗复发性自然流产小鼠的机制研究

姚伟洁¹, 闫仕祺², 袁偲偲¹, 贡磊磊¹, 冯欣^1,*

¹首都医科大学附属北京妇产医院,北京妇幼保健院药事部,北京 100026;
²首都医科大学中医药学院,中医络病研究北京市重点实验室,北京 100069

收稿日期:2024-01-25 出版日期:2024-04-15 发布日期:2024-04-18
通讯作者: ^*冯欣,女,硕士,主任药师,药理学与医院药学。E-mail: fengxin1115@ccmu.edu.cn
作者简介:姚伟洁,男,博士,主管药师,中药药理学。
基金资助:
国家自然科学基金资助项目（82204783）

Study on the mechanism of Yangxue Antai prescription regulating autophagy in the treatment of recurrent spontaneous abortion mice

YAO Weijie¹, YAN Shiqi², YUAN Sisi¹, GONG Leilei¹, FENG Xin^1,*

¹Department of Pharmacy, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing Maternal and Child Health Care Hospital, Beijing 100026, China;
²Beijing Key Laboratory of TCM Collateral Disease Theory Research, School of Traditional Chinese Medicine, Capital Medical University, Beijing 100069, China

Received:2024-01-25 Online:2024-04-15 Published:2024-04-18

摘要/Abstract

摘要： 目的探讨养血安胎方调节自噬影响胎盘血管生成进而治疗复发性自然流产（RSA）的机制。方法对妊娠小鼠分别使用雷帕霉素、3 -甲基腺嘌呤及养血安胎方进行干预,收集小鼠胎盘,苏木素伊红（HE）染色、马松（Masson）染色观察妊娠小鼠的病理改变,免疫组化法测定增殖细胞核抗原（PCNA）、Fibrinogen及CD34蛋白的表达,Western blot法测定血管内皮生长因子（VEGF）、血管内皮细胞生长因子受体2（p-VEGFR2）、p62、Beclin1、LC3Ⅰ和LC3Ⅱ蛋白的表达,透射电镜观察胎盘的超微结构。结果与正常妊娠组比较,雷帕霉素（Rapa）组小鼠取材时体重明显减轻,胚胎吸收率明显增加,HE染色、Masson染色及PCNA与Fibrinogen蛋白表达显示小鼠胎盘发育变差;透射电镜结果与p62、Beclin1和LC3Ⅱ/LC3Ⅰ蛋白表达显示小鼠胎盘自噬水平明显增加;CD34、VEGFA和p-VEGFR2蛋白表达显示胎盘血管生成异常;与Rapa组比较,Rapa+养血安胎组小鼠取材时体重明显增加,胚胎吸收率明显降低,HE染色、Masson染色及PCNA与Fibrinogen蛋白表达显示小鼠胎盘发育得到明显改善,透射电镜结果与p62、Beclin1和LC3Ⅱ/LC3Ⅰ蛋白表达显示小鼠胎盘自噬水平明显降低,CD34、VEGFA和p-VEGFR2蛋白表达显示胎盘血管生成增加。结论养血安胎方通过抑制自噬体和溶酶体的融合,从而抑制自噬,改善妊娠小鼠的胎盘发育状况,促进胎盘的血管生成,从而降低了流产率。

关键词: 复发性自然流产, 自噬, 血管生成, 养血安胎方, 中药复方, 雷帕霉素, 3 -甲基腺嘌呤, 蛋白表达

Abstract: Objective To investigate the mechanism of Yangxue Antai prescription regulating autophagy and affecting placental angiogenesis in the treatment of Recurrent spontaneous abortion (RSA). Methods Rapamycin, 3-methyladenine and Yangxue Antai prescription were used to intervene in pregnant mice, and then the placentas of mice were collected, and the pathological changes of pregnant mice were observed by HE staining and Masson staining; Proliferating Cell Nuclear Antigen (PCNA), Fibrinogen and CD34 protein expression were determined by immunohistochemistry; The expression of vascular endothelial growth factor (VEGF), vascular endothelial growth factor receptor 2 (p-VEGFR2), p62, Beclin1, LC3Ⅰ and LC3Ⅱ were determined by Western blot, and the ultrastructure of the placenta was observed by transmission electron microscopy. Results Compared with the Control group, the body weight in Rapa group decreased significantly, and the embryo absorption rate increased significantly. HE staining, Masson staining and the expression of PCNA and Fibrinogen protein showed poor placental development. The results of transmission electron microscopy(TEM) and the expression of p62, Beclin1 and LC3Ⅱ/LC3Ⅰ protein showed that the level of placental autophagy increased significantly. The expression of CD34, VEGFA and p-VEGFR2 showed abnormal placental angiogenesis. Compared with Rapa group, the body weight of mice in Rapa+Yangxue Antai group increased significantly, and the embryo absorption rate decreased significantly. HE staining, Masson staining, and the expression of PCNA and Fibrinogen protein showed that the placental development of mice improved significantly. The results of TEM and the expression of p62, Beclin1 and LC3Ⅱ/LC3Ⅰ protein showed that the level of placental autophagy decreased significantly, and the expression of CD34, VEGFA and p-VEGFR2 protein showed increased placental angiogenesis. Conclusion Yangxue Antai prescription can inhibit autophagy by inhibiting the fusion of autophagy and lysosome, improve placental development and promote placental angiogenesis in pregnant mice, and thus reduce the abortion rate.

Key words: recurrent spontaneous abortion, autophagy, angiogenesis, Yangxue Antai prescription, herbal compounding, rapamycin, 3-methyladenine, protein expression

中图分类号:

R965

姚伟洁, 闫仕祺, 袁偲偲, 贡磊磊, 冯欣. 养血安胎方调节自噬治疗复发性自然流产小鼠的机制研究[J]. 中国药物警戒, 2024, 21(4): 378-385.

YAO Weijie, YAN Shiqi, YUAN Sisi, GONG Leilei, FENG Xin. Study on the mechanism of Yangxue Antai prescription regulating autophagy in the treatment of recurrent spontaneous abortion mice[J]. Chinese Journal of Pharmacovigilance, 2024, 21(4): 378-385.

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养血安胎方调节自噬治疗复发性自然流产小鼠的机制研究

Study on the mechanism of Yangxue Antai prescription regulating autophagy in the treatment of recurrent spontaneous abortion mice

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