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作 者:杨典凇 潘爽[1] 何丽娜[1] 李艳萍[1] 张琳[1] 牛玉梅[1]
机构地区:[1]哈尔滨医科大学口腔医学院牙体牙髓病科,黑龙江哈尔滨150001
出 处:《口腔医学研究》2016年第4期361-364,共4页Journal of Oral Science Research
基 金:国家自然科学基金(编号:81271132);黑龙江省教育厅科学技术研究项目(编号:12531234);黑龙江省自然科学基金项目(编号:H201440)
摘 要:目的:探究整合素α6对模拟微重力下人牙髓干细胞(human dental pulp stem cells,hDPSCs)粘附能力的影响。方法:以PLGA支架为载体,将采用酶消化法培养的hDPSCs接种在PLGA支架上,分普通重力组和模拟微重力组培养72h,Western blot检测整合素α6(Integrinα6)、整合素αv(Integrinαv)、整合素β1(Integrinβ1)、粘着斑激酶(focal adhesion kinase,FAK)、磷酸化粘着斑激酶(phospho-FAK)的达;si-RNA转染hDPSCs,抑制整合素α6表达,DAPI荧光染色检测转染后hDPSCs在PLGA支架上的粘附数量,Western blot检测转染后hDPSCs中整合素β1、FAK、phospho-FAK的表达。结果:模拟微重力下,整合素α6、phospho-FAK蛋白水平上调(P<0.05),整合素αv、整合素β1和FAK的蛋白水平未见显著性差异;si-RNA转染的hDPSCs粘附能力、phosphoFAK蛋白水平均低于对照组(P<0.05),整合素β1和FAK的蛋白水平未见显著性差异。结论:模拟微重力下,hDPSCs在PLGA支架上粘附能力增强可能与整合素α6及其下游信号分子FAK的表达水平上调相关。Objective:To study the effects of integrinα6on cell adhesion of human dental pulp stem cells seeded on PLGA scaffolds under simulated microgravity.Methods:hDPSCs were seeded on PLGA scaffolds and cultured under the conditions of normal gravity or simulated microgravity for 72 hours.The protein expression level of integrinα6,integrinαv,integrinβ1,FAK and phospho-FAK were measured by Western blot.Integrinα6siRNA was transfected into hDPSCs and DAPI immunofluorescence staining was performed to detect the adhesion ability of hDPSCs.Western blot was conducted to detect the related-protein expression levels.Results:Expression of integrinα6and phospho-FAK were up-regulated in the microgravity group(P〈0.05)compared to control group,while there were no statistical differences in the expression levels of integrinαv,integrinβ1and FAK.Moreover,adhesion ability and phospho-FAK expression of hDPSCs were decreased after transfected with integrinα6siRNA(P〈0.05),while there were no statistical differences in the expression levels of integrinβ1and FAK.Conclusion:Increased adhesion ability of hDPSCs seeded on PLGA scaffolds may be associated with the up-regulation of integrinα6and its downstream molecule FAK.
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