| 余艳丽,陈小波,方海滨,邱 珍,周 斌,夏中元.二十二碳六烯酸对氧糖剥夺环境下大鼠脑星形胶质细胞凋亡及血管生成因子分泌的影响[J].,2019,19(21):4001-4005 |
| 二十二碳六烯酸对氧糖剥夺环境下大鼠脑星形胶质细胞凋亡及血管生成因子分泌的影响 |
| Effects of Docosahexaenoic Acid on the Apoptosis and Secretion of Angiogenic Factors in the Rat Brain Astrocytes under Oxygen-glucose Deprivation Environment |
| 投稿时间：2019-05-09 修订日期：2019-05-31 |
| DOI：10.13241/j.cnki.pmb.2019.21.001 |
| 中文关键词: 二十二碳六烯酸 氧糖剥夺 血管生成因子 星形胶质细胞 |
| 英文关键词: DHA Oxygen-glucose deprivation Angiogenesis Astrocytes |
| 基金项目:国家自然科学基金项目(81671891) |
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| 中文摘要: |
| 摘要 目的：评价二十二碳六烯酸(Docosahexaenoic Acid，DHA)预处理对氧糖剥夺环境下(Oxygen and glucose deprivation，OGD)大鼠脑星形胶质细胞凋亡及血管生成因子分泌的影响。方法：大鼠脑星形胶质细胞传代培养，第3~4代用于实验。采用随机数字法将培养的细胞分为6组：正常对照组、OGD组、OGD+10 μMDHA组、OGD+40 μMDHA组、OGD+10 μMDHA+GW9662组、OGD+40 μMDHA+GW9662组。在所有缺氧模型组(除正常对照组外)先用无糖、无血清的DMEM液置换原培液；其次在OGD+10 μMDHA、OGD+40 μMDHA、OGD+10 μMDHA+GW9662、OGD+40 μMDHA+GW9662组加入相应浓度的DHA，同时在OGD+10 μMDHA+GW9662组和OGD+40μMDHA+GW9662组加入5 μM GW9662(过氧化物酶体增殖物激活受体PPARγ的抑制剂)。预处理完成后，正常对照组和其余各组分别在5% CO2:95%空气和94％N2:5％ CO2:1％O2条件下培养24 h。采用流式细胞技术检测细胞凋亡率，酶联免疫吸附剂测定（Enzyme linked immunosorbent assay，ELISA）检测培养上清液中的促血管生成素1（Angiopoietin-1，Ang1）、促血管生成素2（Angiopoietin2，Ang2）、血管内皮生长因子（vascular endothelial growth factor，VEGF）的分泌量，Western blotting法检测Bax、Bcl-2、caspase-3的表达。结果：与正常对照组比较，其余组细胞凋亡率、Bax、Caspase-3表达水平明显增加(P＜0.01)，而Bcl-2、Bcl-2/Bax表达明显降低(P＜0.01)。与OGD组比较，OGD+10 μMDHA组和OGD+40 μMDHA组细胞凋亡率、Bax、Caspase-3表达水平明显降低(P＜0.01)，而Bcl-2、Bcl-2/Bax表达明显增加(P＜0.01)；Ang1分泌量明显增加(P＜0.01)，而Ang2和VEGF分泌量明显降低(P＜0.01)；上述各指标的差异在OGD+40 μMDHA组里更加显著(P＜0.01)。与OGD组比较，OGD+10 μMDHA+GW9662和OGD+40 μMDHA+GW9662组各个观察指标均无明显差异(P＞0.05)。相关性统计分析结果显示细胞凋亡率与Ang1水平呈显著负相关(P<0.01)，与Ang2和VEGF的水平呈显著正相关(P<0.01)。结论：二十二碳六烯酸(DHA)预处理能够减少大鼠脑星形胶质细胞在氧糖剥夺(OGD)环境下的凋亡，其机制与增加Ang1分泌，减少Ang2和VEGF分泌，进而调控Ang/Tie2信号通路相关。 |
| 英文摘要: |
| ABSTRACT Objective: To evaluate the effects of docosahexaenoic acid(DHA) on the apoptosis and secretion of angiogenic factors in the rat brain astrocytes under oxygen-glucose deprivation(OGD) environment. Methods: Rat astrocytes were cultured in vitro and randomly divided into six groups using a random number table: normal control group, OGD group, OGD+10 μM DHA group, OGD+40 μM DHA group, OGD+10 μM DHA+GW9662, OGD+40 μMDHA+GW9662. The concentration of DHA was 10 μM or 40 μM in the corresponding group according to the design of experiment. In addition, 5 μM GW9662 was added in OGD+10 μM DHA+GW9662 group and OGD+40 μMDHA+GW9662 group. Except of the normal control group that cultured under the condition of 5% CO2:95% air. The other groups were cultured under the condition of 94％N2:5％CO2:1％O2 for 24 h. The cell apoptosis rate was detected with flow cytometry technology. The content of Ang1, Ang2, and VEGF in the culture medium were detected by ELISA.The expression of Bax, Bcl-2, Caspase-3 were detected by Western Blot. Results: Compared with the normal control group, the cell apoptosis rate, expression of Bax, Caspase-3 in the OGD group were significantly increased(P<0.01), and the expression of Bcl-2, Bcl-2/Bax were decreased(P<0.01). Compared with OGD group, the cell apoptosis rate, expression of Bax, and Caspase-3 and the secretion of Ang2 and VEGF were all significantly decreased in the OGD+10 μMDHA group and OGD+40 μMDHA group (P<0.01). and the expression of Bcl-2, Bcl-2/Bax and Ang1 secretion were increased(P<0.01). The difference of the above respective indexes in OGD+40 μMDHA group were more significant than that in the OGD+10 μMDHA group(P<0.01). Compared with OGD group, all measurements showed no significant statistical difference in OGD+10 μM DHA+GW9662 group and OGD+40 μMDHA+GW9662 group(P＞0.05). The cell apoptosis rate has positive correlation with the level of Ang2 and VEGF(P<0.01), but negative correlation with the level of Ang1(P<0.01). Conclusion: Pretreatment with DHA could reduce the apoptosis of rat brain astrocytes under OGD environment, it may be related to regulate the Ang/Tie2 signal pathway by increasing the secretion of Ang1, and reducing the secretion of Ang2 and VEGF. |
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