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作 者:杨迎桂[1] 哈小琴[1] 高瞻[2] 王娟[1] 杨志华[1] 张菊林[1] 林静[1]
机构地区:[1]兰州军区兰州总医院检验科,730050 [2]山东省淄博市中心医院检验科,255000
出 处:《中国医药》2016年第12期1867-1871,共5页China Medicine
基 金:国家自然科学基金(81273568)
摘 要:目的 探讨组蛋白去乙酰化酶1(SIRT1)对脐静脉内皮细胞(HUVEC)增殖、迁移、凋亡和管状结构形成等生物学特性的影响。方法 通过实验确定p人类免疫缺陷病毒(HIV)7-U6-shSIRT1-红色荧光蛋白(RFP)干涉慢病毒(shSIRT1组)及pHIV7-U6-Scramble-RFP对照慢病毒(RFP-SC组)的最佳感染复数,并感染HUVEC,48 h后测定HUVEC内的SIRT1基因和蛋白表达水平;检测HUVEC的增殖能力、迁移能力、凋亡情况及体外成管能力。结果 慢病毒感染HUVEC 48 h后,shSIRT1组的SIRT1基因及蛋白表达水平均明显低于RFP-SC组[(0.271±0.005)比(1.000±0.018),(0.499±0.028)比(1.000±0.023)](P〈0.01)。细胞贴壁的第2、3、4天,shSIRT1组HUVEC的增殖能力明显低于RFP-SC组[(4.84±0.61)比(6.16±1.19)、(6.23±0.35)比(9.67±0.86)、(7.32±0.67)比(10.40±0.61)](均P〈0.01)。shSIRT1组细胞的迁移能力明显低于RFP-SC组[5个视野的平均细胞数:(72±17)个比(195±32)个](P〈0.001)。shSIRT1组的细胞凋亡率明显高于RFP-SC组[(26.9±3.9)%比(9.9±2.0)%](P〈0.05)。shSIRT1组细胞的成管能力明显低于RFP-SC组[5个复孔的平均成管数:(13.9±0.8)个比(25.6±1.4)个](P〈0.05)。结论 HUVEC内SIRT1表达水平下降可以抑制细胞增殖、迁移和体外成管能力,并促进细胞凋亡。Objective To investigate the effects of sirtuin (silent mating type information regulation 2 homolog) 1(SIRT1) on proliferation, migration, apoptosis and tube formation properties of human umbilical vein endothelial cells(HUVECs). Methods Several groups of HUVECs were infected by p-human immunodeficiency virus(HIV)-7-U6-shSIRT1-red fluorescent protein(RFP) slow virus and pHIV7-U6-Scramble-RFP slow virus to determine the optimal Multiplicity of Infection(MOI). After 48 h infection by pHIV7-U6-shSIRT1-RFP(shSIRT1 group) and pHIV7-U6-Scramble-RFP(RFP-SC group) with the optimal MOI, SIRT1 gene and protein expressions, proliferation, migration, apoptosis and tube formation properties of HUVECs were determined. Results SIRT1 gene and protein expressions in shSIRT1 group were significantly lower than those in RFP-SC group after 48 h infection[(0.271±0.005) vs (1.000±0.018), (0.499±0.028) vs (1.000±0.023)](P〈0.01). Proliferation abilities of HUVECs in shSIRT1 group were significantly lower than those in RFP-SC group on day 2, 3, 4 of cell adherence[(4.84±0.61) vs (6.16±1.19),(6.23±0.35) vs (9.67±0.86),(7.32±0.67) vs (10.40±0.61)](P〈0.01). The migration ability[median cell numbers of 5 fields: (72±17) vs (195±32)](P〈0.001) and the tube formation ability[median tube formation numbers of 5 holes: (13.9±0.8) vs (25.6±1.4)](P〈0.05) in shSIRT1 group were significantly lower than those in RFP-SC group. The apoptosis rate in shSIRT1 group was significantly higher than that in RFP-SC group[(26.9±3.9)% vs (9.9±2.0)%](P〈0.05). Conclusions Low expression of SIRT1 in HUVECs can inhibite proliferation, migration, tube formation and promote cell apoptosis.
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