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原花青素对人脐静脉内皮细胞功能及miR-221表达的影响
作者:邹燕 吴斐 季昳弛 何雪梅 周翔宇 
单位:646000  四川泸州  西南医科大学附属医院甲状腺外科(邹燕、吴斐、季昳弛、周翔宇) 646000  四川泸州  西南医科大学附属医院医学实验中心(何雪梅) 
关键词:原花青素 人脐静脉内皮细胞 miR-221 
分类号:R543
出版年,卷(期):页码:2018,43(2):120-125
摘要:

[摘要目的 探讨原花青素(PC)对人脐静脉内皮细胞(HUVECs)增殖、迁移的影响,初步探讨PC作用机制。方法  体外培养HUVECs,采用不同浓度(5255075100μg/ml)PC处理细胞24h后,用CCK-8法筛选出50μg/ml PC做后续实验,继续用细胞划痕实验法检测细胞迁移能力;利用实时荧光定量PCR检测HUVECsmiR-221的表达变化;miRWalk数据库预测miR-221靶基因并对靶基因行GO分析和KEGG Pathway分析。结果 PC处理HUVECs 24h后,与对照组相比,PC 5μg/ml组细胞增殖活性变化不明显(P>0.05),而PC 255075100μg/ml组细胞增殖活性与PC呈浓度依赖性下降(P<0.01);与对照组相比,PC 50μg/ml组迁移能力明显减弱(P<0.01);与对照组相比,50μg/mlmiR-221的表达明显升高(P<0.01)GO分析发现miR-221的靶基因主要与细胞的增殖、迁移、基因的翻译等相关,与细胞增殖、迁移有关的靶基因有ADAM17KITPDGFD等。KEGG Pathway分析发现miR-221的靶基因富集FoxOPI3K-Akt5条信号通路。结论  低浓度PCHUVECs增殖活性无明显影响,一定浓度PC可以抑制HUVECs的增殖和迁移,其机制可能涉及miR-221的上调及FoxOPI3K-Akt等信号通路的参与。

[Abstract]  Objective  To study the effect of procyanidine (PC) on the proliferation and migration of human umbilical vein endothelial cells (HUVECs), determine the expression changes of miR-221, and to investigate the mechanism involved. Methods  HUVECs were cultured in vitro and treated with PC (5, 25, 50, 75, 100μg/ml) for 24 hours, and a PC concentration of 50μg/ml was screened by CCK-8 assay for the follow up experiment, then the cell proliferation activity was detected by the wound healing assay. The expression of miR-221 in HUVECs was detected by real-time quantitative PCR; MiR-221 target genes were predicted in miRWalk database, and the target genes were analyzed by GO and KEGG pathway. Results  Compared with the control group, the HUVECs treated with PC for 24h, their proliferation activity in PC 5μg/ml group did not change obviously (P>0.05), and in PC 25, 50, 75 and 100μg/ml groups decreased in a concentration dependent manner (P<0.01). Compared with the control group, the migration ability of PC 50μg/ml group decreased markedly (P<0.01). Compared with the control group, the expression of miR-221 increased after treatment with PC 50μg/ml (P<0.01). Go analysis indicated that the target genes of miR-221 were mainly related to cell proliferation, migration, gene translation and so on. The target genes related to cell proliferation and migration were ADAM17, KIT, PDGFD and so on. KEGG pathway analysis showed that the target genes of miR-221 enriched 5 signal pathways, such as FoxO, PI3K-Akt and so on. Conclusions  Low concentration of PC has no effect on the proliferation activity of HUVECs. A certain concentration of PC can inhibit the proliferation and migration of HUVECs, of which the mechanism may be involved with the up-regulation of miR-221 and FoxO, PI3K-Akt and other signaling pathways.

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