肿瘤坏死因子-α预处理人脐带间充质干细胞的条件培养基对人脐静脉内皮细胞增殖、凋亡的影响

发布时间：2018-04-23 05:00

本文选题：肿瘤坏死因子α + 间质干细胞　；参考：《中华损伤与修复杂志(电子版)》2016年06期

【摘要】：目的探讨100 ng/m L肿瘤坏死因子-α(TNF-α)预处理人脐带间充质干细胞(h UCMSC)的条件培养基对人脐静脉内皮细胞(HUVEC)增殖、凋亡的影响。方法本实验按随机数字表法将HUVEC分为对照组、条件培养基组和预处理条件培养基组。首先使用含有100 ng/m L TNF-α的内皮细胞培养基刺激HUVEC 12 h后,对照组更换为原培养基,条件培养基组则更换为h UCMSC的条件培养基,以及预处理条件培养基组更换为TNF-α预处理h UCMSC的条件培养基。采用MTT法测定HUVEC的增殖活性,流式细胞仪检测细胞的增殖周期,AO-EB染色和流式细胞仪定性、定量检测细胞的凋亡情况。数据比较采用重复测量方差分析。结果 MTT的检测结果提示,在培养24、48、72 h时,对照组细胞的吸光度值是0.51、0.43、0.32,而条件培养基组和预处理条件培养基组吸光度值是0.56、0.51、0.45和0.59、0.56、0.52。流式细胞仪检测细胞增殖周期(G2/M+S期)的结果示:在培养24、48和72 h时,对照组细胞处于G2/M+S期的比例是(22.34±1.03)%、(10.66±1.07)%、(9.58±0.82)%,而条件培养基组和预处理条件培养基组处于G2/M+S期的比例则分别是(27.95±1.85)%、(18.59±1.48)%、(13.02±1.91)%和(31.17±1.29)%、(22.44±2.28)%、(16.28±1.11)%,3组比较差异有统计学意义(F=58.793,P0.05)。此外,流式细胞仪定性检测结果表明:在培养24、48和72 h时,对照组细胞的凋亡率是(18.95±1.01)%、(16.66±1.51)%、(11.37±1.01)%,而条件培养基组和预处理条件培养基组细胞的凋亡率分别是(12.04±1.13)%、(10.88±1.39)%、(6.88±1.63)%和(8.16±1.44)%、(6.97±1.34)%、(2.97±1.44)%,3组比较差异有统计学意义(F=119.372,P0.05)。AO-EB定性检测细胞凋亡的结果与流式定量检测的结果趋势类似。结论 TNF-α预处理h UCMSC的条件培养基可发挥显著促进HUVEC的增殖和抑制HUVEC凋亡的作用。
[Abstract]:Objective to investigate the effect of human umbilical cord mesenchymal stem cells pretreated with 100 ng/m L tumor necrosis factor- 伪 (TNF- 伪) on the proliferation and apoptosis of human umbilical vein endothelial cells (HUVECs). Methods HUVEC was randomly divided into three groups: control group, conditioned medium group and pretreatment conditioned medium group. At first, the endothelial cell medium containing 100 ng/m L TNF- 伪 was used to stimulate HUVEC for 12 h, then the control group was replaced by the original medium, and the conditioned medium group was replaced by the conditional medium of h UCMSC. The condition medium was replaced by TNF- 伪 pretreatment medium for h UCMSC. The proliferative activity of HUVEC was measured by MTT assay, AO-EB staining by flow cytometry and qualitative analysis by flow cytometry, and apoptosis was detected quantitatively. The data were compared by repeated measurement ANOVA. Results the results of MTT showed that the absorbance value of the control group was 0.51 ~ 0.43 ~ 0.32, while that of the conditioned medium group and the pretreatment medium group was 0.566 ~ 0.51 ~ 0.45 and 0.599 ~ 0.56 ~ 0.62 respectively. Flow cytometry was used to detect the G _ 2 / M _ S phase of cell proliferation cycle. The percentage of cells in G _ 2 / M _ S phase in the control group was 22.34 卤1.03 and 10.66 卤1.07 卤9.58 卤0.82, respectively, while that in the conditioned medium group and the conditioned medium group was 27.95 卤1.855.It was 18.59 卤1.4813.02 卤1.91g% and 22.44 卤2.287.28 卤1.117m respectively in the conditioned medium group and in the conditioned medium group. There was significant difference between the two groups. In addition, the qualitative results of flow cytometry showed that: at 24 h and 72 h of culture, The apoptosis rate of the control group was 16.66 卤1.51, while that of the conditioned medium group and the conditioned medium group was 12.04 卤1.13 + 1.39 卤6.88 卤1.63% and 8.16 卤1.44 卤6.97 卤1.44 respectively. There were significant differences between the three groups: F119.372P0.05N. AO-EB and F119.372P0.05N. The trend is similar. Conclusion the conditioned medium pretreated with TNF- 伪 for h UCMSC can significantly promote the proliferation of HUVEC and inhibit the apoptosis of HUVEC.
【作者单位】：内蒙古医科大学;内蒙古医科大学第三附属医院烧伤科内蒙古烧伤研究所;

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