Agrin对体外损伤大鼠星形细胞迁移的作用

发布时间：2018-05-19 23:14

本文选题：集聚蛋白 + 星形细胞　；参考：《华中科技大学》2011年硕士论文

【摘要】：目的初步探讨外源性agrin对体外机械性损伤的大鼠大脑皮质星形细胞迁移的影响。方法 1.大鼠大脑皮质星形细胞原代培养、纯化及传代。 2.机械性划伤的复制和星形细胞迁移能力的测定:一次传代的单层培养星形细胞采用移液器蓝色枪头作“十”字形划伤,以迁移入划伤区星形细胞数(Na)和星形细胞距划伤边缘最大迁移距离(Dmax),作为评价agrin对星形细胞迁移影响的指标。 3.观察划伤后0h、6h、12h、24h、48h、72h时间点体外培养星形细胞的Na和Dmax值,确定划伤后星形细胞迁移活跃时间段。应用不同浓度的外源性agrin (0ng/ml、5ng/ml、10ng/ml、20ng/ml、50ng/ml、100ng/ml),于划伤后60h观察对损伤星形细胞Na和Dmax的影响。在此基础上,选定agrin作用浓度组(5ng/ml、10ng/ml、20ng/ml)及伤后不同时间点(24h、48h、60h),观察agrin对星形细胞迁移影响。Dmax测定用IPP6.0软件,所有数据均应用SPSS13.0软件进行统计学分析。结果 1.划伤72h星形细胞的Na和Dmax值均较伤后48h增加最显著(P0.0001)。 2.划伤后60h外源性agrin 5ng/ml组、10ng/ml组及20ng/ml组均较对照组星形细胞Na和Dmax值均有显著增加(P0.05)。100ng/ml组,星形细胞的Na和Dmax则显著减少(P0.05)。 3.外源性agrin 5ng/ml、10ng/ml、20ng/ml浓度组和在伤后24h、48h、60h,星形细胞的Na和Dmax均较对照组显著增加(P0.05),以10ng/ml浓度组对于星形细胞迁移影响最为明显(P0.05)。结论 1.星形细胞在机械性划伤12h后开始出现迁移,划伤后48h-72h星形细胞迁移能力明显增加; 2.外源性agrin的加入可影响损伤星形细胞的迁移。Agrin在一定浓度(5~20ng/ml)和作用时间范围(24~48h)内可显著促进损伤星形细胞迁移;但过高的agrin作用浓度(如50~100ng/ml)则抑制星形细胞迁移。
[Abstract]:Objective to investigate the effect of exogenous agrin on the migration of astrocytes in rat cerebral cortex after mechanical injury in vitro. Method 1. Primary culture, purification and passage of astrocytes from rat cerebral cortex. 2. Replication of mechanical scratches and determination of the ability of stellate cells to migrate: a single passage of astrocytes was cut in the form of "ten" zigzag by using a liquidator with a blue gun head. The effect of agrin on astrocyte migration was evaluated by the maximum migration distance between the astrocytes and the edge of the scratched area (Na) and the maximum migration distance between the astrocytes and the edge of the scratches. 3. The Na and Dmax values of cultured astrocytes were observed at 0 h, 6 h, 12 h, 24 h, 48 h and 72 h after scratch, and the active period of astrocyte migration was determined. The effects of different concentrations of exogenous agrin on the Na and Dmax of astrocytes were observed at 60 h after scratch with 10 ng / ml 10 ng / ml 20 ng / ml 50 ng / ml / ml 100 ng / ml 路ml / ml of different concentrations of exogenous agrin / mg / ml 5 ng / ml ~ 5 ng / ml ~ (-1) ~ 10 ng / ml ~ (-1) ~ 20 ng / ml ~ (-1). On this basis, 5 ng / ml 10 ng / ml of agrin and 20 ng / ml ~ 20 ng / ml) and 24 h ~ 48 h ~ (-1) ~ 60 h after injury were selected to observe the effect of agrin on the migration of astrocytes. IPP6.0 software was used to determine the effect of agrin on the migration of astrocytes. All the data were analyzed statistically by SPSS13.0 software. Result 1. The Na and Dmax values of astrocytes were significantly increased at 72 h compared with 48 h after injury (P 0.0001). 2. The Na and Dmax values of astrocytes in 10 ng / ml group and 20ng/ml group were significantly higher than those in control group at 60 h after scratch, while Na and Dmax in astrocytes decreased significantly compared with those in control group. 3. Na and Dmax of astrocytes in exogenous agrin 5 ng / ml 10 ng / ml 20 ng / ml group and at 24 h after injury at 48 h or 60 h after injury were significantly increased compared with the control group, especially in the 10ng/ml group, which had the most significant effect on the migration of astrocytes. Conclusion 1. The migration of astrocytes began to occur 12 hours after mechanical scratch, and the migration ability of 48h-72h astrocytes increased significantly after scratching. 2. The addition of exogenous agrin could affect the migration of injured astrocytes. Agrin could significantly promote the migration of injured astrocytes within a certain concentration of 20 ng / ml and within a time range of 244h, but too high concentration of agrin (such as 50 ~ 100ng / ml) inhibited the migration of astrocytes.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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