脊髓损伤后早期IL-1Β通过JAK2-STAT3信号通路促进胶质瘢痕形成

发布时间：2018-04-10 10:13

本文选题：脊髓损伤　切入点：IL-1β　引自：《重庆医科大学》2017年硕士论文

【摘要】：目的探讨IL-1β对脊髓损伤后胶质纤维酸性蛋白(glial fibrillary acidic protein,GFAP)及波形蛋白(Vimentin)表达的影响及JAK2-STAT3信号通路在其中发挥的作用。方法(1)建立大鼠急性脊髓损伤模型,按不同的干预措施将大鼠随机分为模型组(model group)、假手术组(sham group)、IL-1β特异性抑制剂组(IL-1RA group)、IL-1β组(IL-1βgroup)。分别于建模后大鼠的脊髓损伤处给予生理盐水、IL-1RA、IL-1β各10ul,假手术组只打开椎板,不作其他处理。各组大鼠在术后相应时间点(术后8h、12h、1天、3天、7天、14天)利用BBB评分测定其运动障碍程度,运用Western blotting、免疫荧光、免疫组化等技术观察各处理组间相关指标(GFAP、p-STAT3)的变化趋势。(2)建立大鼠急性脊髓损伤模型,按不同的干预措施将大鼠随机分为模型组(model group)、假手术组(sham group)、IL-1β+JAK2-STAT3特异性抑制剂组(IL-1β+AG490 group)、IL-1β组(IL-1βgroup)。分别于建模后大鼠的脊髓损伤处给予生理盐水、IL-1β+AG490、IL-1β各10ul,假手术组只打开椎板,不作其他处理。各组大鼠在术后相应时间点(术后1、3、7、14天)利用BBB评分测定其运动障碍程度,运用Western blotting、免疫荧光等技术检测各处理组间相关指标(GFAP、Vimentin)的变化趋势。结果(1)术后第8h、12h,IL-1β特异性抑制剂组p-STAT3表达相比模型组显著降低(p0.05),IL-1β组p-STAT3表达相比模型组显著升高(p0.05),各处理组p-STAT3表达相比假手术组均显著升高(p0.01)。术后第7、14天,IL-1β特异性抑制剂组GFAP表达相比模型组显著降低(p0.05),IL-1β组GFAP表达相比模型组显著升高(p0.05),各处理组GFAP表达相比假手术组均显著升高(p0.01)。术后第14天,IL-1β特异性抑制剂组BBB评分相比模型组显著提高(p0.05),IL-1β组BBB评分相比模型组显著降低(p0.05),各处理组BBB评分相比假手术组均显著降低(p0.01)。术后第1、3天,各组GFAP表达相较模型组无显著差异;术后第1、3、7天,各组BBB评分相较模型组无显著差异(p0.05)。术后第7、14天,IL-1β+AG490组GFAP、Vimentin表达相比模型组显著降低(p0.05),IL-1β组GFAP、Vimentin表达相比模型组显著升高(p0.05),各处理组GFAP、Vimentin表达相比假手术组均显著升高(p0.01)。术后第14天,IL-1β+AG490组BBB评分相比模型组显著提高(p0.05),IL-1β组BBB评分相比模型组显著降低(p0.05),各处理组BBB评分相比假手术组均显著降低(p0.01)。术后第1、3天,各组GFAP、Vimentin表达相较模型组无显著差异;术后第1、3、7天,各组BBB评分相较模型组无显著差异(p0.05)。结论急性脊髓损伤后分泌的IL-1β可通过磷酸化JAK2-STAT3促进损伤局部胶质瘢痕形成,进而影响大鼠后肢运动功能。
[Abstract]:Objective to investigate the effect of IL-1 尾 on the expression of glial fibrillary acidic protein (GFAP) and vimentin (Vimentin) after spinal cord injury (sci) and the role of JAK2-STAT3 signaling pathway.Methods A rat model of acute spinal cord injury was established. Rats were randomly divided into two groups according to different intervention measures: model group, sham group, IL-1 尾 specific inhibitor group, IL-1RA group and IL-1 尾 group.The rats were given normal saline IL-1RA-IL-1 尾 at the spinal cord injury area after modeling, and the sham operation group only opened the lamina without any other treatment.The degree of motor dysfunction was measured by BBB score, Western blotting and immunofluorescence were used to measure the degree of motor dysfunction at the corresponding time points (8 h, 12 h, 1 day, 3 days and 14 days after operation).The rats were given normal saline IL-1 尾 AG490 and IL-1 尾 10 ul. the sham operation group only opened the lamina without any other treatment.BBB scores were used to measure the degree of motor dysfunction at the corresponding time points (1 day 3 days after operation), and Western blotting, immunofluorescence and other techniques were used to detect the change trend of the relative indexes of each treatment group.Results (1) the expression of p-STAT3 in IL-1- 尾 specific inhibitor group was significantly lower than that in model group at 8 h after operation. Compared with model group, the expression of p-STAT3 in IL-1 尾 group was significantly higher than that in model group. The p-STAT3 expression in each treatment group was significantly higher than that in sham operation group.The expression of GFAP in the IL-1 尾 specific inhibitor group was significantly lower than that in the model group on the 7th day after operation. Compared with the model group, the expression of GFAP in the IL-1 尾 group was significantly higher than that in the model group. The GFAP expression in each treatment group was significantly higher than that in the sham operation group.On the 14th day after operation, the BBB score of IL-1 尾 specific inhibitor group was significantly higher than that of model group. Compared with model group, the BBB score of IL-1 尾 group was significantly lower than that of model group, and the BBB score of each treatment group was significantly lower than that of sham operation group.On the 1st day after operation, the expression of GFAP in each group was not significantly different from that in the model group, but on the 3rd day after operation, there was no significant difference in BBB score between each group and the model group (P 0.05).On the 14th day after operation, the BBB score of IL-1 尾 AG490 group was significantly higher than that of model group. Compared with model group, the BBB score of IL-1 尾 group was significantly lower than that of model group. The BBB score of each treatment group was significantly lower than that of sham operation group.On the 1st day after operation, there was no significant difference in the expression of Vimentin in each group as compared with the model group, but on the 3rd day after operation, there was no significant difference in BBB score between each group and the model group (p0.05).Conclusion IL-1 尾 secreted after acute spinal cord injury can promote the formation of local glial scar by phosphorylated JAK2-STAT3, and then affect the motor function of hindlimb in rats.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R651.2

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