摘要
目的研究FGF2对缺氧诱导的巩膜成纤维细胞(scleral fibroblasts,SF)增殖和胶原的影响并探讨其可能调节的下游信号通路。方法用5%O2刺激SF 24 h诱导近视SF模型,RT-qPCR检测FGF2 mRNA表达,Western blot检测FGF2蛋白表达。细胞计数试剂盒8(Cell Count Kit-8,CCK-8)、流式细胞术和Western blot分别检测细胞增殖活力、细胞凋亡和胶原代谢相关蛋白collagenⅠ、MMP2以及通路蛋白PERK、p-PERK、EIF2α、EIF2α、ATF4表达。结果缺氧刺激可促进FGF2 mRNA和蛋白表达升高(P<0.01)并激活PERK/EIF2α/ATF4通路(P<0.001),抑制SF细胞增殖(P<0.001)和collagenⅠ表达(P<0.001),诱导MMP2表达(P<0.001)及细胞凋亡(P<0.001)。敲降FGF2或经PERK抑制剂GSK2606414处理可逆转缺氧对SF细胞的作用,促进细胞增殖活力(P<0.001)和collagenⅠ表达(P<0.01),减少细胞凋亡(P<0.01)。结论FGF2通过调节PERK/EIF2α/ATF4通路的激活,影响缺氧诱导的SF增殖及胶原代谢。
Objectives To investigate the effects of FGF2 on the proliferation and collagen production of hypoxia-induced scleral fibroblasts(SF)and explore the downstream signaling pathways it regulates.Methods 5%O2 was used to stimulate the SF to induce myopia SF model for 24 hours.RT-qPCR was used to detect FGF2 mRNA expression,and Western blot analysis was used to check FGF2 protein expression.The Cell Counting Kit-8(CCK-8),flow cytometry,and Western blot were used to assess cell proliferation vitality,cell apoptosis,and the expression of collagen metabolism-related proteins collagen I,MMP2,and pathway proteins PERK,p-PERK,EIF2α,EIF2α,and ATF4.Results Hypoxia increased FGF2 mRNA and protein expression(P<0.01),activated the PERK/EIF2α/ATF4 pathway(P<0.001),inhibited SF cell proliferation(P<0.001)and collagen I expression(P<0.001),while induced MMP2 expression(P<0.001)and apoptosis(P<0.001).Knocking down FGF2 or treating with PERK inhibitor GSK2606414 reversed the effect of hypoxia on SF cells,increased cell proliferation(P<0.001)and collagenⅠexpression(P<0.01),and suppressed cell apoptosis(P<0.01).Mechanism study revealed that FGF2 knockdown dampened the activation of PERK/EIF2α/ATF4 pathway.Conclusion FGF2 affects hypoxia-induced SF proliferation and collagen metabolism by regulating the activation of PERK/EIF2α/ATF4 signaling pathway.
作者
翟瑜如
白艳
李云云
ZHAI Yuru;BAI Yan;LI Yunyun(Dept.of Ophthalmology,Changzhi People’s Hospital,Changzhi Shanxi 046000,China)
出处
《昆明医科大学学报》
CAS
2024年第10期22-28,共7页
Journal of Kunming Medical University
基金
山西省卫健委基金资助项目(2018132)。
