SDF-1通过PI3K/Akt和MAPK/Erk信号通路对人微血管内皮细胞功能产生影响
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新疆维吾尔自治区自然科学基金(2014211C129)


SDF-1 affects human microvascular endothelial cell function by influence PI3K/Akt and MAPK/Erk signaling pathway
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    摘要:

    目的:观察基质细胞衍生因子-1(stromal cell derived factor-1,SDF-1)对人微血管内皮细胞 (human microvascular endothelial cell line-1,HMEC-1)功能的影响,并对相关机制进行初步探讨,从而明确SDF-1在糖尿病血管病变中的作用。方法:体外培养HMEC-1,分别在培养基中加入不同浓度的SDF-1(0-25-50-100 -滋g/L),Western blot检测HMEC-1中PI3K/Akt和MAPK/Erk信号通路的活化情况,MTT和流式细胞分析检测HMEC-1增殖和凋亡能力的变化情况,划痕愈合实验检测HMEC-1迁移能力的变化情况。采用PI3K/Akt和MAPK/Erk信号通路抑制剂LY294002和U0126阻断HMEC-1中PI3K/Akt和MAPK/Erk信号通路后,再以SDF-1处理HMEC-1,MTT和流式细胞分析检测HMEC-1增殖和凋亡能力的变化情况;划痕愈合实验检测HMEC-1迁移能力的变化情况。结果:Western blot结果显示,25 -滋g/L的SDF-1处理即可显著活化HMEC-1中的PI3K/Akt和MAPK/Erk信号通路,且随着SDF-1处理浓度的升高,PI3K/Akt和MAPK/Erk信号通路活化水平逐渐升高。同0 -滋g/L 的SDF-1处理组相比,25-50-100 -滋g/L的SDF-1均可显著加强HMEC-1的增殖和迁移能力(P < 0.01),并抑制HMEC-1的凋亡水平(P < 0.01),且这种作用具有剂量依赖性。当采用LY294002和U0126阻断HMEC-1中的PI3K/Akt和MAPK/Erk信号通路后,SDF-1促HMEC-1增殖和迁移能力及抑制HMEC-1凋亡的能力均显著降低(P < 0.01)。结论:SDF-1可通过活化PI3K/Akt和MAPK/Erk信号通路,进而促进HMEC-1的增殖和迁移,并抑制HMEC-1的凋亡水平,从而在糖尿病血管病变中发挥一定的作用。

    Abstract:

    Objective:To observe the effect of stromal cell derived factor-1 (SDF-1)on human microvascular endothelial cell line-1(HMEC-1),and study the relevant mechanism. Methods:HMEC-1 cells were treated with SDF-1 of different concentrations(0,25,50,100 -滋g/L),then the phosphorylation level of PI3K/Akt and MAPK/Erk were detected by Western blot,and the proliferation ability and apoptosis rate of HMEC-1 cells were detected by MTT and FCM. Changes on migration ability of HMEC-1 cells was detected by wound healing assay. MCF-7 cells were pretreated with PI3K/Akt and MAPK/Erk signaling pathway inhibitor to observe the effect of PI3K/Akt and MAPK/Erk signaling pathway on SDF-1 induced proliferation,apoptosis and migration in HMEC-1 cells. Results:Compared with 0 -滋g/L SDF-1 treated group,25,50 and 100 -滋g/L SDF-1 treatment actived PI3K/Akt and MAPK/Erk signaling pathway,promoted proliferation and migration ability,and induced HMEC-1 cells apoptosis(P < 0.01),and those effects were concentration-dependent. When pretreated with PI3K/Akt and MAPK/Erk signaling pathway inhibitor,the effect of SDF-1 on promoting proliferation and migration ability,and inducing HMEC-1 cells apoptosis ability were significantly blocked. Conclusion:By activation PI3K/Akt and MAPK/Erk signaling pathway,SDF-1 can promote proliferation and migration ability,and induce HMEC-1 cells apoptosis,thus plays a role in diabetic angiopathy.

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高 静,朱红霞,王敏哲,苟 静,张 丽. SDF-1通过PI3K/Akt和MAPK/Erk信号通路对人微血管内皮细胞功能产生影响[J].南京医科大学学报(自然科学版),2015,(9):1205-1210

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  • 收稿日期:2015-01-13
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  • 在线发布日期: 2015-09-10
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