养血安胎方含药血清调节外泌体释放影响内皮细胞及子宫内膜细胞的机制研究OA
Study on the Mechanism of Medicated Serum from Yangxue Antai Formula in Influencing Endothelial and Endometrial Cells via Regulating Exosome Release
目的:探讨养血安胎方含药血清调节人绒毛膜癌(JEG-3)细胞外泌体的释放从而影响内皮细胞及子宫内膜细胞的机制.方法:使用外泌体释放抑制剂GW4869 抑制JEG-3 细胞外泌体的释放,养血安胎方含药血清干预JEG-3 细胞,并建立JEG-3 细胞或其分泌的外泌体与人子宫内膜腺癌(HEC-1-A)细胞或人脐静脉内皮细胞(HUVECs)的共培养体系,采用Western blot法测定HUVECs的血管内皮生长因子A(VEGFA)及磷酸化血管内皮生长因子受体 2(p-VEGFR2)的表达,HEC-1-A 细胞中上皮-间质转化(EMT)相关蛋白上皮型钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)、神经型钙黏蛋白(N-cadherin)及纤连蛋白(Fibronectin)的表达,以及蜕膜化相关蛋白胰岛素样生长因子结合蛋白 1(IGFBP-1)、催乳素(PRL)、同源框基因A10(HOXA10)及骨形态发生蛋白2(BMP2)的表达,细胞划痕实验测定HUVECs的细胞迁移率,Transwell小室实验测定HUVECs的侵袭与迁移能力,细胞骨架实验测定HUVECs的纤维状肌动蛋白(F-actin)表达,小管生成实验测定HUVECs的管腔长度.结果:与空白对照组比较,GW4869 组中VEGFA及p-VEGFR2 的表达显著降低(P<0.01);E-cadherin表达显著增加(P<0.01),Vimentin、N-cadherin和Fibronectin的表达显著降低(P<0.01);BMP2、HOXA10、PRL和IGFBP1 的表达显著降低(P<0.01);细胞迁移和侵袭能力显著降低(P<0.01),管腔长度显著降低(P<0.01),F-actin 表达显著降低(P<0.01).与 GW4869 组比较,GW4869+养血安胎方组和GW4869+外泌体组中VEGFA及p-VEGFR2 的表达显著增加(P<0.01);E-cadherin表达显著降低(P<0.01),Vimentin、N-cadherin和Fibronectin的表达显著增加(P<0.01);BMP2、HOXA10、PRL和IGFBP1 的表达显著增加(P<0.05,P<0.01);细胞迁移和侵袭能力显著增加(P<0.01),管腔长度显著增加(P<0.01),F-actin表达显著增加(P<0.01).结论:养血安胎方含药血清通过调节JEG-3 细胞外泌体的释放,从而影响内皮细胞的血管生成,以及子宫内膜细胞EMT及蜕膜化过程.
Objective:To investigate the mechanism of Yangxue Antai serum in regulating the release of exosomes from JEG-3 cells to affect endothelial cells and endometrial cells.Methods:The exosome release inhibitor GW4869 was used to inhibit the release of exosomes from JEG-3 cells,and the Yangxue Antai serum interfered with JEG-3 cells,A co-culture system was established in which JEG-3 cells or exosomes secreted by JEG-3 cells were co-cultured with HEC-1-A cells or HUVECs.The expression of VEGFA and p-VEGFR2 in HUVECs,the expression of EMT-related proteins E-cadherin,Vimentin,N-cadherin and Fibronectin as well as the expression of deciduation-related proteins BMP2,HOXA10,PRL and IGFBP1 in HEC-1-A cells were measured by Western blot.The migration rate of HUVECs was measured by cell scratch assay.The invasion and migration ability of HUVECs were measured by Transwell assay,F-actin expression was measured by cytoskeleton assay,and the lumen length in HUVECs was measured by tubule formation assay.Results:Compared with Control group,the expressions of VEGFA and p-VEGFR2 in GW4869 group were significantly decreased.The expression of E-cadherin was significantly increased,while the expression of Vimentin,N-cadherin and Fibronectin were significantly decreased.The expressions of BMP2,HOXA10,PRL and IGFBP1 were significantly decreased.Cell invasion and migration ability were significantly decreased,the lumen length was decreased,and the expression of F-actin was significantly decreased.Compared with GW4869 group,the expressions of VEGFA and p-VEGFR2 in GW4869+Yangxue Antai and GW4869+exosome group were significantly increased.The expression of E-cadherin was significantly decreased,while the expression of Vimentin,N-cadherin and Fibronectin was significantly increased.The expressions of BMP2,HOXA10,PRL and IGFBP1 were significantly increased.Cell invasion and migration ability were increased significantly,the lumen length was increased,and the expression of F-actin was increased significantly.Conclusion:Yangxue Antai serum can affect the angiogenesis of endothelial cells,EMT and decidualization of endometrial cells by regulating the release of exosomes from JEG-3 cells.
姚伟洁;冯欣
首都医科大学附属北京妇产医院/北京妇幼保健院,北京 100006||首都医科大学基础-临床联合实验室,北京 100006首都医科大学附属北京妇产医院/北京妇幼保健院,北京 100006||首都医科大学基础-临床联合实验室,北京 100006
医药卫生
外泌体血管生成上皮细胞-间质转化蜕膜化
ExosomeAngiogenesisEpithelial-mesenchymal transformationDecidualization
《中医药学报》 2026 (3)
6-14,9
国家自然科学基金项目(82204783)
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