B6-Co小鼠眼睑成纤维细胞增殖、凋亡及迁移能力实验研究
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摘要
目的探讨B6-Co小鼠出生时眼睑闭合不全的形成机制。方法分离、培养并鉴定B6-Co小鼠及其背景品系C57BL/6J(B6)眼睑成纤维细胞,CCK8法检测B6-Co、 B6小鼠眼睑成纤维细胞的增殖水平,Annexin V-FITC法检测B6-Co、B6小鼠眼睑成纤维细胞的凋亡水平,Transwell检测B6-Co、B6小鼠眼睑成纤维细胞的迁移能力。结果成功培养出B6-Co、B6小鼠眼睑成纤维细胞,B6-Co小鼠眼睑成纤维细胞的增殖能力和迁移能力极显著低于B6小鼠(P<0.01),B6-Co小鼠眼睑成纤维细胞的凋亡水平极显著高于B6小鼠(P<0.01)。结论 B6-Co小鼠眼睑成纤维细胞的增殖能力、凋亡水平及迁移能力均与B6小鼠有极显著差异。
Objective To explore the mechanism of eyelid incomplete closure at birth in C57 BL/6 Jcorneal opacity(B6-Co) mice. Methods The eyelid fibroblasts of B6 and B6-Co mice were isolated,cultured and identified. CCK8 method was used to detect the proliferation of eyelid fibroblasts in B6 and B6-Co mice. Apoptosis of eyelid fibroblasts in B6 and B6-Co mice was detected by Annexin V-FITC.Transwell was used to detect the migration ability of eyelid fibroblasts in B6 and B6-Co mice.Results The eyelid fibroblasts of B6 and B6-Co mice were successfully cultured. The proliferation and migration of eyelid fibroblasts from B6-Co mice were significantly lower than those of B6 mice(P<0.01). The apoptosis of eyelid fibroblasts in B6-Co mice was significantly higher than that of B6 mice(P<0.01). Conclusion The proliferative capacity, apoptosis level and migration ability of eyelid fibroblasts in B6-Co mice were significantly different from those of B6 mice.
Objective To explore the mechanism of eyelid incomplete closure at birth in C57 BL/6 Jcorneal opacity(B6-Co) mice. Methods The eyelid fibroblasts of B6 and B6-Co mice were isolated,cultured and identified. CCK8 method was used to detect the proliferation of eyelid fibroblasts in B6 and B6-Co mice. Apoptosis of eyelid fibroblasts in B6 and B6-Co mice was detected by Annexin V-FITC.Transwell was used to detect the migration ability of eyelid fibroblasts in B6 and B6-Co mice.Results The eyelid fibroblasts of B6 and B6-Co mice were successfully cultured. The proliferation and migration of eyelid fibroblasts from B6-Co mice were significantly lower than those of B6 mice(P<0.01). The apoptosis of eyelid fibroblasts in B6-Co mice was significantly higher than that of B6 mice(P<0.01). Conclusion The proliferative capacity, apoptosis level and migration ability of eyelid fibroblasts in B6-Co mice were significantly different from those of B6 mice.
引文
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[12]Binelli A,Magni S,La Porta C,et al.Cellular pathways affected by carbon nanopowder-benzo(alpha)pyrene complex in human skin fibroblasts identified by proteomics.l[J].Ecotox Environ Safe,2018,160:144-153.
[13]Sano K,Someya T,Hara K,et al.Effect of traditional plants in Sri Lanka on skin fibroblast cell number[J].Data Brief,2018,19:611-615.
[14]Curtain M,Heffner CS,Maddox DM,et al.A novel allele of Alx4 results in reduced Fgf10 expression and failure of eyelid fusion in mice[J].Mamm Genome,2015,26(3-4):173-180.
[15]Lee CC,Hsieh TS.Wuho/WDR4 deficiency inhibits cell proliferation and induces apoptosis via DNA damage in mouse embryonic fibroblasts[J].Cell Signal,2018,47:16-26.
[16]Liu Y,Geng YH,Yang H,et al.Extracellular ATP drives breast cancer cell migration and metastasis via S100A4 production by cancer cells and fibroblasts[J].Cancer lett,2018,430:1-10.
[2]陈家祺.我国角膜病的研究现状和展望[J].现代医院,2002,2(3):6-8.
[3]邵义祥,吴宝金,薛整风,等.遗传性角膜基质变性小鼠及其突变基因的定位[J].南京师范大学学报:自然科学版,2006,29(2):99-102.
[4]尹依恒.乙烷基亚硝基脲诱变眼睑开放小鼠模型角膜组织学及突变基因研究[D].扬州:扬州大学,2018.
[5]Tao H,Ono K,Kurose H,et al.Exogenous FGF10 can rescue an eye-open at birth phenotype of Fgf10-null mice by activating activin and TGFalpha-EGFR signaling[J].Dev Growth Differ,2006,48(5):339-346.
[6]吴刘成,刘春,蒋荧梅,等.角膜混浊小鼠突变候选基因Map3k1克隆与序列分析[J].动物医学进展,2011,32(4):1-5.
[7]Wu LC,Liu C,Jiang MR,et al.Defective eyelid leading edge cell migration in C57BL/6-corneal opacity mice with an“eye open at birth”phenotype[J].Genet Mol Res,2016,15(3).doi:10.4238/gmr.15036741.
[8]卢泽艳,郑良凤,李瑶,等.B6-Co与B6小鼠眼睑角质形成细胞的原代无血清培养及比较研究[J].湖北农业科学,2013,52(2):393-397.
[9]Cressey A,Jacobs DS,Remington C,et al.Improvement of chronic corneal opacity in ocular surface disease with prosthetic replacement of the ocular surface ecosystem(PROSE)treatment[J].Am J Ophth Case Rep,2018,10:108-113.
[10]Bozkurt B,Yildirim MS,Okka M,et al.GAPO syndrome:Four new patients with congenital glaucoma and myelinated retinal nerve fiber[J].Am J Med Genet A,2013,161A(4):829-834.
[11]Chen VM,Shelke R,Nystrom A,et al.Collagen VII deficient mice show morphologic and histologic corneal changes that phenotypically mimic human dystrophic epidermolysis bullosa of the eye[J].Exp Eye Res,2018,175:133-141.
[12]Binelli A,Magni S,La Porta C,et al.Cellular pathways affected by carbon nanopowder-benzo(alpha)pyrene complex in human skin fibroblasts identified by proteomics.l[J].Ecotox Environ Safe,2018,160:144-153.
[13]Sano K,Someya T,Hara K,et al.Effect of traditional plants in Sri Lanka on skin fibroblast cell number[J].Data Brief,2018,19:611-615.
[14]Curtain M,Heffner CS,Maddox DM,et al.A novel allele of Alx4 results in reduced Fgf10 expression and failure of eyelid fusion in mice[J].Mamm Genome,2015,26(3-4):173-180.
[15]Lee CC,Hsieh TS.Wuho/WDR4 deficiency inhibits cell proliferation and induces apoptosis via DNA damage in mouse embryonic fibroblasts[J].Cell Signal,2018,47:16-26.
[16]Liu Y,Geng YH,Yang H,et al.Extracellular ATP drives breast cancer cell migration and metastasis via S100A4 production by cancer cells and fibroblasts[J].Cancer lett,2018,430:1-10.