硫酸镁对大鼠神经干细胞辐射损伤保护作用的初步研究
发布时间：2019-01-05 04:36
【摘要】： 目的:研究硫酸镁对神经干细胞辐射损伤的保护作用。 方法:取新生24h内的SD大鼠脑组织,进行神经干细胞的原代培养并进行鉴定。将神经干细胞分为空白对照组、实验对照组和实验用药组,分别用2Gy、4Gy ~(60)Coγ射线对实验组及实验对照组进行照射,照射后24h、48h、72h用CCK-8检测神经干细胞的增殖,流式细胞仪检测凋亡及周期变化,透射电镜观察细胞核的形态变化,并与实验对照组进行比较。用SAS统计软件包进行数据处理。 结果: 1.分别用丝裂元进行神经干细胞培养传代后, EGF培养的神经干细胞主要分化为GFAP阳性的神经胶质细胞。而EGF及bFGF联合进行培养的神经干细胞,分化为NSE阳性的神经元细胞比例明显较单用EGF组增多,GFAP阳性神经胶质细胞也明显减少。 2.在神经干细胞增殖情况的研究中,发现不同浓度的硫酸镁对神经干细胞增殖情况有不同的影响。当浓度大于5mg/ml时将导致神经干细胞的死亡,当其浓度为1.25mg/ml、2.5mg/ml时,硫酸镁将能促进神经干细胞的增殖。 3.将神经干细胞传代后,分别用2Gy及4Gy ~(60)Coγ射线照射神经干细胞,照射后24h、48h、72h时间点发现,实验对照组与空白对照组比较其增殖明显降低(p0.05),实验用药组与实验对照组比较其增殖情况明显好转,除24h时间点外,各时间点比较均有统计学意义(p0.05)。 4.神经干细胞经2Gy、4Gy ~(60)Coγ射线照射后24h、48h、72h研究发现,与空白对照组比较实验对照组其凋亡率明显增加(p0.05),细胞核固缩、核仁边集,而各时间点实验用药组与实验对照组比较,其凋亡率明显好转(p0.05),细胞核形态明显好转,更接近空白对照组。 5.神经干细胞经2Gy、4Gy ~(60)Coγ射线照射后培养24h、48h、72h,与空白对照组比较,实验对照组细胞G1期阻滞明显增加(p0.05),同时G2期也有轻微的阻滞,且4Gy组较2Gy组,G2期阻滞更明显。实验用药组与实验对照组比较,各时间点G1期阻滞明显降低(p0.05),更接近空白对照组。 结论: 1. EGF具有促进神经干细胞向胶质细胞分化的趋势。bFGF与EGF同时培养时,能使神经干细胞分化为神经元的比例增加。 2.当硫酸镁的浓度为1.25mg/ml、2.5mg/ml时能促进神经干细胞增殖。 3.早期运用硫酸镁可阻止电离辐射对神经干细胞增殖的损伤;使电离辐射所致神经干细胞凋亡率降低,对电离辐射所致神经干细胞损伤起到保护作用;同时能够降低电离辐射所致神经干细胞G1期阻滞,减轻电离辐射对神经干细胞的损伤。
[Abstract]:Objective: to study the protective effect of magnesium sulfate on radiation injury of neural stem cells. Methods: the brain tissues of SD rats within 24 hours were collected and the primary culture and identification of neural stem cells were carried out. Neural stem cells were divided into blank control group, experimental control group and experimental drug group. The experimental group and experimental control group were irradiated with 2 Gy ~ (4) Gy ~ (60) Co 纬 -ray, respectively. The proliferation of neural stem cells was detected by CCK-8 at 24 h, 48 h and 72 h after irradiation. Apoptosis and cycle changes were detected by flow cytometry and morphological changes of nucleus were observed by transmission electron microscope and compared with experimental control group. The data are processed by SAS statistical software package. Results: 1. After the neural stem cells were cultured and subcultured with mitogen, the neural stem cells cultured with EGF mainly differentiated into GFAP positive glial cells. However, the proportion of neural stem cells differentiated into NSE positive neurons in EGF and bFGF co-culture group was significantly higher than that in EGF group alone, and the number of GFAP positive glial cells was also significantly decreased. 2. In the study of the proliferation of neural stem cells, it was found that different concentrations of magnesium sulfate had different effects on the proliferation of neural stem cells. When the concentration is higher than 5mg/ml, the neural stem cells will die. When the concentration is 1.25 mg / ml or 2.5 mg / ml, magnesium sulfate can promote the proliferation of neural stem cells. 3. The neural stem cells were irradiated with 2Gy and 4Gy ~ (60) Co 纬 -rays respectively. The proliferation of neural stem cells in the experimental control group was significantly lower than that in the blank control group (p0.05). Compared with the experimental control group, the proliferation of the experimental group was improved obviously, except 24 h time point, the comparison of each time point had statistical significance (p0.05). 4. Compared with the control group, the apoptosis rate of NSCs was significantly increased (p0.05), nuclear pyknosis and nucleolar edge aggregation were observed in the experimental control group after 24 h ~ (48 h) irradiation with 2 Gy ~ (4) Gy ~ (60) Co 纬 -ray. Compared with the control group, the apoptotic rate and nuclear morphology of the experimental drug group were significantly improved (p0.05), which were closer to the blank control group. 5. Neural stem cells were irradiated with 2 Gy ~ (4) Gy ~ (60) Co 纬 -ray for 24 h or 48 h and 72 h after irradiation. Compared with the control group, the G _ 1 phase arrest was significantly increased in the experimental control group (p0.05), and the G _ 2 phase was also slightly blocked in the 4Gy group compared with the 2Gy group. G 2 arrest was more obvious. Compared with the experimental control group, the G1 phase block in the experimental group was significantly decreased (p0.05), which was closer to the blank control group. Conclusion: 1. EGF could promote the differentiation of neural stem cells into glial cells. When bFGF and EGF were cultured at the same time, the proportion of neural stem cells differentiated into neurons was increased. 2. When the concentration of magnesium sulfate is 1.25 mg / ml, 2.5 mg / ml, it can promote the proliferation of neural stem cells. 3. Early application of magnesium sulfate could prevent the proliferation of neural stem cells induced by ionizing radiation, decrease the apoptosis rate of neural stem cells induced by ionizing radiation, and play a protective role on the injury of neural stem cells induced by ionizing radiation. At the same time, it can reduce the G 1 arrest of neural stem cells induced by ionizing radiation, and alleviate the injury of neural stem cells induced by ionizing radiation.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R363
本文编号：2401279
[Abstract]:Objective: to study the protective effect of magnesium sulfate on radiation injury of neural stem cells. Methods: the brain tissues of SD rats within 24 hours were collected and the primary culture and identification of neural stem cells were carried out. Neural stem cells were divided into blank control group, experimental control group and experimental drug group. The experimental group and experimental control group were irradiated with 2 Gy ~ (4) Gy ~ (60) Co 纬 -ray, respectively. The proliferation of neural stem cells was detected by CCK-8 at 24 h, 48 h and 72 h after irradiation. Apoptosis and cycle changes were detected by flow cytometry and morphological changes of nucleus were observed by transmission electron microscope and compared with experimental control group. The data are processed by SAS statistical software package. Results: 1. After the neural stem cells were cultured and subcultured with mitogen, the neural stem cells cultured with EGF mainly differentiated into GFAP positive glial cells. However, the proportion of neural stem cells differentiated into NSE positive neurons in EGF and bFGF co-culture group was significantly higher than that in EGF group alone, and the number of GFAP positive glial cells was also significantly decreased. 2. In the study of the proliferation of neural stem cells, it was found that different concentrations of magnesium sulfate had different effects on the proliferation of neural stem cells. When the concentration is higher than 5mg/ml, the neural stem cells will die. When the concentration is 1.25 mg / ml or 2.5 mg / ml, magnesium sulfate can promote the proliferation of neural stem cells. 3. The neural stem cells were irradiated with 2Gy and 4Gy ~ (60) Co 纬 -rays respectively. The proliferation of neural stem cells in the experimental control group was significantly lower than that in the blank control group (p0.05). Compared with the experimental control group, the proliferation of the experimental group was improved obviously, except 24 h time point, the comparison of each time point had statistical significance (p0.05). 4. Compared with the control group, the apoptosis rate of NSCs was significantly increased (p0.05), nuclear pyknosis and nucleolar edge aggregation were observed in the experimental control group after 24 h ~ (48 h) irradiation with 2 Gy ~ (4) Gy ~ (60) Co 纬 -ray. Compared with the control group, the apoptotic rate and nuclear morphology of the experimental drug group were significantly improved (p0.05), which were closer to the blank control group. 5. Neural stem cells were irradiated with 2 Gy ~ (4) Gy ~ (60) Co 纬 -ray for 24 h or 48 h and 72 h after irradiation. Compared with the control group, the G _ 1 phase arrest was significantly increased in the experimental control group (p0.05), and the G _ 2 phase was also slightly blocked in the 4Gy group compared with the 2Gy group. G 2 arrest was more obvious. Compared with the experimental control group, the G1 phase block in the experimental group was significantly decreased (p0.05), which was closer to the blank control group. Conclusion: 1. EGF could promote the differentiation of neural stem cells into glial cells. When bFGF and EGF were cultured at the same time, the proportion of neural stem cells differentiated into neurons was increased. 2. When the concentration of magnesium sulfate is 1.25 mg / ml, 2.5 mg / ml, it can promote the proliferation of neural stem cells. 3. Early application of magnesium sulfate could prevent the proliferation of neural stem cells induced by ionizing radiation, decrease the apoptosis rate of neural stem cells induced by ionizing radiation, and play a protective role on the injury of neural stem cells induced by ionizing radiation. At the same time, it can reduce the G 1 arrest of neural stem cells induced by ionizing radiation, and alleviate the injury of neural stem cells induced by ionizing radiation.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R363
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