摘要
目的观察大鼠急性脊髓损伤(Acute Spinal Cord Injury,ASCI)后神经细胞炎症反应、自噬和凋亡相关因子的变化,探讨靶向干预JAK2/STAT3信号通路活化的治疗作用。方法选择健康成年8~10周龄SD雄性大鼠共45只,随机分为假手术组、模型组和JAK2抑制剂AG-490干预组各15只,采用改良Allen法复制模型。分别于6、12、24 h处死大鼠(每组每个时间点各5只),采用免疫组织化学染色法检测脊髓组织白细胞介素6(IL-6)、肿瘤坏死因子-α(TNF-α)和热休克蛋白-27(Heat Shock Protein-27,HSP-27)阳性表达,Western Blot法检测组织JAK2、p-JAK2、STAT3和p-STAT3蛋白表达,免疫荧光标记法检测组织微管相关蛋白Ⅰ轻链3(Microtubule-Associated Protein1 Light Chain 3,MAP1LC3)-Ⅱ阳性表达,Western Blot法检测组织Caspase-3和Bax/Bcl-2蛋白表达。结果与假手术组比较,模型组各时间点组织IL-6、TNF-α和HSP-27阳性表达率,p-JAK2和p-STAT3蛋白表达水平,MAP1LC3-Ⅱ阳性表达率以及Caspase-3、Bax/Bcl-2蛋白表达水平均有所增加,而干预组各时间点上述指标均低于模型组(P<0.05)。结论ASCI早期病理改变可能涉及神经细胞的炎症反应、自噬和凋亡以及JAK2/STAT3信号通路的活化,靶向干预JAK2/STAT3信号通路的活化可能为ASCI的治疗提供新的方向。
Objective The study aimed to explore the inflammation,autophagy,apoptosis and JAK2/STAT3 signaling pathway in neurons after Acute Spinal Cord Injury(ASCI)rats.Methods A total of 45 healthy adult 8-10 week male SD rats with an average weight of 245 g were randomly divided into sham operation group,model group and JAK2 inhibitor AG-490 intervention group with 15 rats in each group,and the model was reproduced by the modified Allen method.The rats were killed at 6h,12 h and 24 h respectively(5 rats in each group at each time point).IL-6,TNF-αand Hsp-27 were detected by immunohistochemistry,the expressions of JAK2,p-JAK2,STAT3 and p-STAT3 proteins were detected by Western Blot,the expressions of LC3-Ⅱwas detected by immunofluorescence,caspase-3 and Bax/Bcl-2 proteins were detected by Western Blot.Results Compared with the sham operation group,the positive expression rates of IL-6,TNF-αand Hsp-27,levels of p-JAK2 and p-STAT3 proteins,LC3-Ⅱpositive expression rate,levels of caspase-3 and Bax/Bcl-2 proteins were significantly higher in the model group at each time points,while the above indexes in the intervention group were significantly lower than those in the mode group(P<0.05).Conclusion Early pathological changes of ASCI may be related to inflammatory response,autophagy,apoptosis and activation of JAK2/STAT3 signaling pathway in neural cells of early ASCI.
作者
彭成
刘佩雷
陶钧
张乐
陶奇昌
代建昊
李强
PENG Cheng;LIU Peilei;TAO Jun;ZHANG Le;TAO Qichang;DAI Jianhao;LI Qiang(Department of orthopedices,the First Affiliated Hospital Anhui University ofScience Technology/the First People′s Hospital of Huainan,Huainan Anhui 232000,China)
出处
《转化医学杂志》
2021年第2期83-88,共6页
Translational Medicine Journal
基金
淮南市科技计划项目(201040600903)。
