摘要
目的:探究温经汤含药血清对异位子宫内膜细胞增殖、迁移与凋亡的影响和作用机制。方法:以Wistar雌性大鼠来制备空白血清、温经汤含药血清以及阳性药物含药血清,并处理子宫内膜上皮细胞系End1/E6E7。免疫荧光检测增殖细胞核抗原(PCNA)和切割型胱天蛋白酶-3(Cleaved Caspase-3)表达;流式细胞术测定细胞周期和细胞凋亡;划痕试验测定细胞迁移;免疫印记测定Cleaved Caspase-3表达;荧光定量PCR测定lncRNA-H19与miR-216a-5p mRNA表达;双荧光素酶检测lncRNA-H19与miR-216a-5p的结合关系;RNA免疫沉淀(RIP)实验观察lncRNA-H19与miR-216a-5p的结合能力。结果:与空白血清组比较,温经汤含药血清和阳性药物含药血清组细胞活性降低(P<0.01),其中10%温经汤含药血清抑制效果最佳。温经汤含药血清组和阳性药物含药血清干预后,细胞中PCNA表达、细胞迁移率以及miR-216a mRNA表达降低,而G_(0)/G_(1)期细胞比例、Cleaved Caspase-3表达、细胞凋亡率以及lncRNA-H19 mRNA表达升高,与空白血清组比较差异均有统计学意义(均P<0.01)。双荧光素酶和RIP实验证实lncRNA-H19能够与miR-216a-5p结合并抑制其表达。过表达LncRNA-H19后,温经汤含药血清处理End1/E6E7细胞中的lncRNA-H19 mRNA表达水平升高(P<0.01),miR-216a mRNA表达水平降低(P<0.01)。此外,过表达lncRNA-H19逆转了温经汤含药血清对End1/E6E7细胞增殖、迁移以及凋亡的影响。结论:温经汤含药血清通过调控lncRNA-H19/miR-216a轴来抑制子宫内膜异位细胞增殖与迁移,诱导细胞凋亡。
Objective:To investigate the effect and mechanism of Wenjing Decoction-containing serum on the proliferation,migration,and apoptosis of ectopic endometrial cells.Methods:Female Wistar rats were used to prepare blank serum,Wenjing Decoction-containing serum,and positive drug-containing serum to treat the endometrial epithelial cell line End1/E6E7.Immunofluorescence was used to detect proliferating cell nuclear antigen(PCNA) and Cleaved Caspase-3 expression.Flow cytometry was employed to determine cell cycle and apoptosis.Scratch assay was conducted to determine cell migration.Immunoblotting was applied to determine Cleaved Caspase-3 expression.Fluorescence quantitative PCR was utilized to determine lncRNA-H19 and miR-216a-5p mRNA expression.Dual Luciferase was used to detect the binding relationship between lncRNA-H19 and miR-216a-5p.RNA immunoprecipitation(RIP) assay was performed to observe the binding ability of lncRNA-H19 and miR-216a-5p.Results:Compared with the blank serum group,the cellular activity was decreased in Wenjing Decoction-containing serum and positive drug-containing serum groups(P<0.01),of which the serum containing 10% of Wenjing Decoction had the best inhibitory effect.After the intervention of Wenjing Decoction-containing serum and positive drug-containing serum,PCNA expression,cell migration rate and miR-216a mRNA expression were decreased in cells,while the cell proportion in G_(0)/G_(1) phase,Cleaved Caspase-3 expression,apoptosis rate,and lncRNA-H19 mRNA expression were increased,which were significantly different from that in the blank serum group(P<0.01).Dual Luciferase and RIP assays confirmed that lncRNA-H19 was able to bind to miR-216a-5p and inhibit its expression.After LncRNA-H19 overexpression,the lncRNA-H19 mRNA expression level was increased in End1/E6E7 cells treated with Wenjing Decoction-containing serum(P<0.01),while the miR-216a mRNA expression level was decreased(P<0.01).In addition,lncRNA-H19 overexpression reversed the effects of Wenjing Decoction-containing serum on the proliferation,migration,and apoptosis of End1/E6E7 cells.Conclusion:Wenjing Decoction-containing serum inhibits proliferation and migration and induces apoptosis of endometriotic cells by regulating the lncRNA-H19/miR-216a axis.
作者
李潇
王娜梅
马丽亚
姚曼曼
张晓宇
张雪琳
张大伟
LI Xiao;WANG Namei;MA Liya;YAO Manman;ZHANG Xiaoyu;ZHANG Xuelin;ZHANG Dawei(The Second Clinical College,Henan University of Chinese Medicine,Zhengzhou 450002,China;The Third Clinical College,Henan University of Chinese Medicine,Zhengzhou 451100,China;Henan University of Chinese Medicine,Zhengzhou 450000,China)
出处
《世界中医药》
北大核心
2025年第6期906-914,共9页
World Chinese Medicine
基金
国家自然科学基金面上项目(82074562)
2023年河南省科技攻关项目(232102310478)。
