摘要
摘要
目的：
探讨AMD3100通过CXCR4/β-catenin信号通路对中枢性神经干细胞增殖、迁移和分化的抑制作用。
方法：
1、悬浮法培养中枢神经干细胞，配制含AMD3100不同浓度梯度（0ug/L，40ug/L, 80ug/L, 160ug/L, 320ug/L, 640ug/L和1000ug/L）的培养基培养神经干细胞。分别在2小时、24小时、48小时、72小时用CCK8法检测各浓度梯度OD值，再分别计算各时间时的增殖抑制率。
2、根据前一步AMD3100抑制中枢神经干细胞增殖实验结果选择最适药物浓度为80ug/L和最佳作用时间为48小时。分别培养对照组（0ug/L）和AMD3100组（80ug/L）中枢神经干细胞48小时，划痕实验检测细胞迁移能力的变化，Western Blot检测神经干细胞标志物Nestin、神经元细胞标志物β-tubulin、Wnt、β-catenin和β-catenin（P-Tyr489）的表达情况。
结果：
1、CCK8法检测结果：各浓度梯度的AMD3100作用于神经干细胞后，随着时间的延长，80ug/L, 160ug/L, 320ug/L, 640ug/L和1000ug/L各浓度下增殖能力逐渐降低，抑制率逐渐增高，在48小时和72小时，增殖能力均极显著低于0ug/L，差别具有统计学意义（P＜0.01），抑制率均达到50%以上；40ug/L浓度时增殖能力与0ug/L相比无明显差异（P>0.05）。在2小时和24小时，随着浓度的增加各浓度梯度抑制率逐渐增加；在48小时和72小时160ug/L, 320ug/L和640ug/L抑制率呈增加趋势，均低于80ug/L和1000ug/L，均高于24小时各浓度梯度抑制率。
2、划痕实验结果：在48小时AMD3100组细胞迁移面积百分比为(19.80%±1.83)，明显低于对照组（32.92%±2.64），差异具有统计学意义（P ＜0.05）。Western Blot检测48小时AMD3100组Nestin、β-tubulin、β-catenin 和β-catenin（P-Tyr489）的表达量均低于与对照组，差异均有统计学意义（P ＜0.05）。两组Wnt的表达量无明显差异（P>0.05）。
摘要
结论：
1、AMD3100抑制中枢神经干细胞的增殖具有时间和浓度依赖性。
2、AMD3100抑制SDF-1/CXCR4轴可以抑制中枢神经干细胞的迁移和分化能力。
什么伞下雨不能用
3、AMD3100可能是通过CXCR4/β-catenin通路抑制中枢神经干细胞的增殖迁移和分化。
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蜘蛛图片大全大图关键词：神经干细胞；AMD3100；CXCR4；Wnt/β-catenin信号通路
Abstract
ABSTRACT
Objective:
To investigate the inhibition effect of AMD3100 on the proliferation, migration and differentiation of central neural stem cells by CXCR4/β-catenin signaling pathway.
Methods:
1.Preparation of AMD3100 containing different concentrations (0ug/L, 40ug/L, 80ug/L, 160ug/L, 320ug/L, 640ug/L and 1000ug/L) in the culture medium.Central neural stem cells was cultured by suspension method. The OD value of each group was detected by CCK8 method at 2 hours, 24 hours, 48 hours and 72 hours respectively, and the proliferation inhibition rate of each group was calculated at different times.
2.The drug concentration and reaction time (80ug/L, 48h) were lected from the previous experiment. The control group (0ug/L) and AMD3100 group (80ug/L) cells were cultured for 48 hours. Wound Healing was ud to detect the migration ability of central neural stem cells and Western Blot was ud to detect the expression of Nestin, β-tubulin, Wnt, β-catenin and β-catenin (P-Tyr489). Results:
1.CCK8 assay results: With the increa of the extension time, in 80ug/L, 160ug/L, 320ug/L, 640ug/L, and 1000ug/L, the proliferation capacity was gradually decread and the inhibition rate was gradually incread. At 48 hours and 72 hours, the proliferation capacity was significantly lower than that of 0 ug/L (P <0.01), and the inhibition rate of all was more than 50%.The cell proliferation ability of 40ug/L was no significant difference compared with 0ug/L(P> 0.05).At 2 hours and 24 hours, the inhibition rate was gradually incread with increasing concentration. At 48 hours and 72 hours, the inhibition rates of 160ug/L, 320ug/L and 640ug/L with a increasing trend were lower than 80ug/L and 1000ug/L, respectively, but higher than all concentrations of 24 hours.
Abstract
2.The Wood Healing results: At 48 hours, percentage of the area of cell migration in AMD3100 group
(19.80% ±1.83) was significantly lower than the control group(32.92% ± 2.64), the difference was statistically significant (P<0.05). Western Blot results: The expression of Nestin, β-tubulin, β-catenin and β-catenin (P-Tyr489) in AMD3100 group was lower than that in control group (P<0.05).There was no significant difference in Wnt expression between the two groups (P> 0.05). Conclusion:
1.AMD3100 inhibit the proliferation of neural stem cells in a time and concentration-dependent.关于励志的手抄报
2. AMD3100 could inhibit the proliferation, migration and differentiation of neural stem cells.
3. AMD3100 may be through CXCR4/β-catenin pathway to inhibit the proliferation and migration and differentiation of neural stem cells.
Key Words:neural stem cells; AMD3100; CXCR4; Wnt/β-catenin signaling pathway.
目录
目录自制卤水豆腐
第1章引言 (1)
第2章中枢神经干细胞培养及AMD3100对其增殖能力的影响 (3)
情何归
阻断药去哪里买2.1 材料 (3)
2.1.1中枢神经干细胞 (3)
2.1.2主要试剂及AMD3100 (3)
2.1.3主要实验仪器 (3)
2.1.4 主要试剂的配制 (4)
2.2 实验方法 (5)
2.2.1中枢神经干细胞的培养、传代及冻存 (5)
2.2.2 CCK8法检测各浓度梯度的AMD3100对中枢神经干细胞增殖能力
的影响 (6)
2.3 数据处理及统计学分析 (7)
2.4 结果 (7)
2.4.1 中枢神经干细胞培养结果 (7)
金华双龙洞
2.4.2 不同AMD3100浓度梯度对中枢神经干细胞增殖能力的影响 (8)
2.4.3不同AMD3100浓度梯度中枢神经干细胞增殖抑制率的变化 (8)
2.5 结论 (9)
第3章AMD3100对中枢神经干细胞迁移分化的实验研究 (10)
3.1 材料 (10)
3.1.1主要试剂 (10)
3.1.2主要实验仪器 (11)
3.1.3 主要试剂的配制 (11)
3.2 实验方法 (13)
3.2.1 划痕实验检测细胞迁移能力 (13)
3.2.2 Western Blot实验检测各组细胞Nestin、β-tubulin、Wnt、β-catenin
和β-catenin（P-Tyr489）的表达 (13)
3.3 数据处理及统计学分析 (17)
