摘要
目的探究c-Jun氨基末端激酶(JNK)信号通路对创伤性颅脑损伤(TBI)后小鼠海马区神经干细胞(NSC)增殖的影响。方法选择36只4~6周龄雄性C57BL/6小鼠随机分为假手术组、模型组(采用控制性皮质损伤方式建立TBI模型)和JNK抑制剂组(建模+腹腔注射JNK信号抑制剂SP600125),采用改良神经功能评分评估各组小鼠神经功能,测定各组小鼠大脑含水量,透射电镜观察海马神经元突触超微结构,免疫荧光染色检测NSC增殖(BrdU/Sox2阳性)情况,免疫印迹检测海马区自噬蛋白(LC3-Ⅱ、Beclin1、Atg5)及JNK信号蛋白表达。结果与假手术组比较,模型组mNSS评分、大脑含水量、海马区BrdU/Sox2双阳性细胞数(30.6±2.8比114.2±3.9)个、LC3-Ⅱ、Beclin1、Atg5、JNK、p-JNK蛋白相对表达量升高;与模型组比较,JNK抑制组mNSS评分、大脑含水量、LC3-Ⅱ、Beclin1、Atg5、JNK、p-JNK蛋白相对表达量降低,海马区BrdU/Sox2双阳性细胞数(137.43±3.40)个数升高(P<0.05);透射电镜下可见,假手术组的海马CA1区神经细胞胞核正常;模型组海马CA1区神经细胞结构出现不同程度扩张或肿胀,细胞核出现融合;JNK抑制剂组的细胞结构出现少量扩张和肿胀,胞核膜轻度融合。结论抑制JNK信号通路能通过降低自噬水平提高TBI小鼠海马区NSC增殖能力,改善神经功能。
Objective To explore the role of c-Jun amino-terminal kinase(JNK)signaling pathway in the proliferation of hippocampal neural stem cells(NSC)in mice with traumatic brain injury(TBI).Methods Thirty-six 4-to 6-week-old male C57BL/6 mice were selected,and randomly assigned into sham group,model group(TBI modeling induced using controlled cortical impact method),and JNK inhibitor group(modeling+intraperitoneal injection of JNK inhibitor SP600125).The neurological function was assessed using modified Neurological Severity Score(mNSS),and brain water content was measured in all three groups.The synaptic ultrastructure of hippocampal neurons was observed by transmission electron microscopy,the proliferation of NSC(BrdU/Sox2-colabeled cells)was detected by immunofluorescence staining,and the expression of autophagy-associated proteins(LC3-Ⅱ,Beclin1,Atg5)and JNK signalling related proteins in the hippocampus was measured by Western blotting.Results Compared with sham group,model group detected elevated mNSS score,increased brain water content,larger number of BrdU/Sox2-colabeled cells in hippocampal regions(30.6±2.8 vs 114.2±3.9),and up-regulated relative expression of LC3-Ⅱ,Beclin1,Atg5,JNK,and p-JNK proteins.The abnormality elevated modified mNSS scores,increased brain water content,and up-regulated relative expression of LC3-Ⅱ,Beclin1,Atg5,JNK,and p-JNK proteins were all attenuated in JNK group compared to model group,and the number of BrdU/Sox2-colabeled cells in hippocampal regions was(137.43±3.40)in JNK inhibitor group,which was larger than that of model group,with statistical difference(P<0.05).Transmission electron microscopy showed that the nucleus of hippocampal CA1 neurons in sham operation group was normal.In model group,the nerve cell structure in the hippocampal CA1 region showed varying degrees of expansion or swelling,and the nucleus appeared fusion.The cell structure in JNK inhibitor group showed a small amount of expansion and swelling,and mild fusion of the nuclear membrane.Conclusion Inhibitio
作者
梁新
李佳庆
王玉保
曹晓光
霍虹达
Liang Xin;Li Jiaqing;Wang Yubao;Cao Xiaoguang;Huo Hongda(Seventh Department of Neurosurgery,Cangzhou Central Hospital,Cangzhou 061000,China)
出处
《中华生物医学工程杂志》
CAS
2024年第3期201-206,共6页
Chinese Journal of Biomedical Engineering
基金
河北省2023年度医学科学研究课题(20232116)
