Page 21 - 南京医科大学学报自然科学版

第43卷第3期 南京医科大学学报（自然科学版）
2023年3月 Journal of Nanjing Medical University（Natural Sciences） ·311 ·

·基础研究·

STK25在星形胶质细胞活化和EAE小鼠病变进程中的作用

王一凡，谢金玉，陈迎玉，邓嘉欣，冯 巧，韩 欣，史 震，周 峰，刘晓梅 *
江苏省免疫与代谢重点实验室，徐州医科大学病原生物学与免疫学教研室，江苏 徐州 221004

［摘 要］ 目的：探讨丝氨酸/苏氨酸蛋白激酶25（serine/threonine protein kinase 25，STK25）对星形胶质细胞活化及实验性自身免
疫性脑脊髓炎（experimental autoimmune encephalomyelitis，EAE）小鼠疾病进程的影响。方法：髓鞘少突胶质细胞糖蛋白35⁃55
（myelin oligodendrocyte glycoprotein 35⁃55，MOG35⁃55 ）诱导小鼠 EAE 模型，Real⁃time PCR、Western blot 和 ELISA 法分别检测脊髓
组织中STK25、肿瘤坏死因子⁃α（tumor necrosis factor⁃α，TNF⁃α）与干扰素诱导蛋白⁃10（interferon⁃inducible protein⁃10，IP⁃10）的
转录水平与蛋白表达；免疫荧光双标法检测STK25在脊髓星形胶质细胞中的表达。干扰素（interferon，IFN）⁃γ体外刺激小鼠原
代星形胶质细胞后不同时间，上述方法检测STK25的蛋白表达及细胞因子的转录水平变化。将特异性靶向星形胶质细胞的对
照及STK25敲减的重组慢病毒悬液感染星形胶质细胞后，IFN⁃γ刺激，测定TNF⁃α与IP⁃10的转录水平；同时给予小鼠病毒悬液
7 d后，诱导EAE模型，Western blot 检测脊髓组织STK25的表达；Real⁃time PCR 检测脊髓炎性因子与趋化因子转录水平。HE
和劳克坚牢蓝染色（Luxol fast blue，LFB）观察脊髓炎症细胞浸润与髓鞘损伤。结果：与对照组相比，EAE组小鼠STK25的表达
水平显著降低，炎性因子与趋化因子的转录与分泌水平明显升高；同时脊髓组织星形胶质细胞STK25的表达明显下降。IFN⁃γ
体外刺激星形胶质细胞STK25表达降低时，炎性因子生成增加；进一步敲减星形胶质细胞内源性的STK25基因，体外星形胶质
细胞及体内EAE小鼠脊髓中炎性因子与趋化因子的生成显著升高，同时加剧了EAE小鼠脊髓炎症细胞浸润与髓鞘脱失。结
论：STK25可能通过影响星形胶质细胞的活化，调节炎性与趋化因子的产生，从而参与EAE小鼠的病变进程。
［关键词］ 丝氨酸/苏氨酸蛋白激酶25；星形胶质细胞；实验性自身免疫性脑脊髓炎；炎性因子
［中图分类号］ R593.2 ［文献标志码］ A ［文章编号］ 1007⁃4368（2023）03⁃311⁃08
doi：10.7655/NYDXBNS20230303

The role of STK25 in astrocyte activation and pathological changes in EAE mice
WANG Yifan，XIE Jinyu，CHEN Yingyu，DENG Jiaxin，FENG Qiao，HAN Xin，SHI Zhen，ZHOU Feng，LIU
Xiaomei *
Jiangsu Key Laboratory of Immunity and Metabolism，Department of Pathogen Biology and Immunology，Xuzhou
Medical University，Xuzhou 221004，China

［Abstract］ Objective：This study aims to investigate the effects of serine/threonine protein kinase 25（STK25）on the activation of
astrocytes as well as the process of experimental autoimmune encephalomyelitis（EAE）in mice. Methods：The EAE mouse model was
induced by myelin oligodendrocyte glycoprotein 35⁃55（MOG35 ⁃ 55 ）polypeptide. The transcription levels and the expression of STK25，
tumor necrosis factor⁃ α（TNF⁃α）and interferon inducible protein 10（IP⁃10）in spinal cord tissue and serum were detected by ELISA，
Real⁃time PCR and Western blot，respectively. The expression of STK25 in astrocytes in spinal cord tissue of mice was detected by
immunofluorescence double staining. Primary mouse astrocytes were stimulated in vitro with interferon⁃γ（IFN⁃γ），and the expression
of STK25 and the transcription levels of cytokines were detected by the above methods at different times. Primary mouse astrocytes
were transfected by recombinant lentiviral vector，and stimulated with IFN ⁃ γ，the transcription levels of TNF ⁃ α and IP ⁃ 10 were
measured. Meantime，the EAE model was induced after injecting vectors 7 days，and the expression of STK25 in spinal cord tissue was
detected by Western blot，the transcription levels of TNF ⁃ α and IP ⁃ 10 were detected by Real ⁃ time PCR，and the infiltration of
inflammatory cells and the demyelination of spinal cord were observed by HE and Luxol fast blue（LFB）staining methods，respectively.
Results：Compared with control group，the transcription level and the expression of STK25 in EAE group were significantly reduced

［基金项目］ 国家自然科学基金（81971179）；江苏省自然科学基金（BK20191463）
∗
通信作者（Corresponding author），E⁃mail：lxmlxm_hi@xzhmu.edu.cn