| 李晓洁,徐玲玲,郑苏,赵峰.高压氧联合跑台运动对脑瘫幼鼠内质网应激、神经元树突棘发育及JAK/STAT信号通路的影响[J].中国康复,2025,40(11):643-649 |
| 高压氧联合跑台运动对脑瘫幼鼠内质网应激、神经元树突棘发育及JAK/STAT信号通路的影响 |
| Effects of hyperbaric oxygen combined with treadmill exercise on endoplasmic reticulum stress, neuronal dendritic spine development and JAK/STAT signaling pathway in young cerebral palsy rats |
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| DOI：10.3870/zgkf.2025.11.001 |
| 中文关键词: 高压氧 跑台运动 脑瘫 内质网应激 神经元树突棘发育 |
| 英文关键词: hyperbaric oxygen treadmill exercise cerebral palsy endoplasmic reticulum stress neuronal dendritic spines development |
| 基金项目:2021-2022年度湖北省卫健委中医药科研立项项目（青年人才项目）（ZY2021Q015） |
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| 中文摘要: |
| 目的：研究高压氧联合跑台运动对脑瘫幼鼠内质网应激、神经元树突棘发育及Janus激酶/信号转导与转录激活子（JAK/STAT）信号通路的影响。方法：将64只幼鼠随机分为N组（正常对照组）、M组（模型组）、H组（模型+高压氧）、R组（模型+跑台运动）、U组（模型+高压氧+跑台运动）5组（N组12只，其余各组各13只）。除N组外均建立缺血缺氧脑瘫幼鼠模型。采用触觉刺激试验检测各组幼鼠行为学改变，记录撕去胶布所用时间。酶联免疫吸附法（ELISA）检测血清中炎性指标肿瘤坏死因子α（TNF-α）、白细胞介素1β（IL-1β）、白细胞介素10（IL-10）水平，免疫荧光染色法检测内质网应激指标蛋白激酶样内质网激酶（PERK）、半胱氨酸天冬氨酸蛋白酶12（caspase-12）蛋白表达，使用高尔基染色试剂盒对海马CA区标本染色后分析顶树突与底树突树突棘密度，HE染色观察脑组织形态学变化和神经元损伤情况，免疫印迹法检测JAK/STAT蛋白表达水平。结果：M组、H组、R组、U组撕除胶布时间长于N组（P<0.05），H组、R组、U组撕除胶布时间短于M组，且U组所用时间最短（P<0.05）。与N组比较，M组、H组、R组、U组血清TNF-α、IL-1β、脑组织PERK、caspase-12、p-JAK2、p-STAT3蛋白表达升高，IL-10蛋白表达降低，CA区底树突棘密度降低（P<0.05），与M组比较，H组、R组、U组TNF-α、IL-1β、PERK、caspase-12、磷酸化的JAK2（p-JAK2）、磷酸化的STAT3（p-STAT3）水平降低，IL-10水平升高且U组最显著，CA区底树突棘密度升高，U组最高（P<0.05）。各组JAK2、STAT3水平、CA区顶树突棘密度比较无显著性差异。H组、R组、U组细胞形态有所改善，细胞染色变浅，炎性浸润减少，神经纤维缠结减轻，其中U组细胞形态改善最明显。与N组比较，M组、H组、R组、U组正常神经细胞数目降低（P<0.05），与M组比较，H组、R组、U组正常神经细胞数目升高，且U组最高（P<0.05）。结论：高压氧与跑台运动对脑瘫幼鼠内质网应激、神经元树突棘发育均有改善作用，抑制JAK/STAT信号通路激活，且两者联合效果更佳。 |
| 英文摘要: |
| Objective: To study the effects of hyperbaric oxygen combined with treadmill running on endoplasmic reticulum stress, neuronal dendritic spine development and JAK/STAT signaling pathway in young cerebral palsy rats. Methods: In total, 64 young rats were randomly divided into N group (normal control group), M group (model group), H group (model+hyperbaric oxygen), R group (model+treadmill exercise), U group (model+hyperbaric oxygen+treadmill exercise) (12 in N group, and 13 in the rest groups). The model of ischemic hypoxic cerebral palsy was established except for N group. The behavioral changes of young mice in each group were detected using the tactile stimulation test, and the time taken to tear off the tape was recorded. Serum levels of tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β) and IL-10 were detected by ELISA. Endoplasmic reticulum stress indexes ［including the protein expression of protein kinase-like endoplasmic reticulum kinase (PERK) and cysteine aspartic protease 12 (Caspase-12)］ were analyzed by immunofluorescence staining. The dendrite density of apical dendrite and basal dendrite was analyzed by staining with Golgi staining kit. Morphological changes of brain tissue and neuronal injury were observed by HE staining. The expression level of JAK/STAT protein was detected by Western blotting. Results: The time of tearing off the tape was longer, and IL-10 level was lower in M group, H group, R group and U group than those in N group (P<0.05). The time of tearing off the tape in H group, R group and U group was shorter than that in M group M, and that in U group was the shortest (P<0.05). Compared with N group, the expression levels of serum TNF-α, IL-1β, brain PERK, Caspase-12, P-JAK2 and P-Stat3 protein in M, H, R and U groups were increased, and IL-10, the density of basal dendrite spines in CA region was decreased (P<0.05). The levels of TNF-α, IL-1β, PERK, Caspase-12, P-JAK2 and P-Stat3 in H, R and U groups were decreased, the lowest in U group, IL-10, the density of basal dendrite spines in CA region was increased, and the highest in U group (P<0.05). There were no significant differences in JAK2 and STAT3 levels and apical dendrite spine density in CA region among all groups. The cell morphology of H, R and U groups was improved, the cell staining became shallow, the inflammatory infiltration was reduced, and the nerve fiber tangles were alleviated, among which the cell morphology of U group showed the most obvious improvement. Compared with N group, the number of normal nerve cells in M, H, R, and U groups decreased (P<0.05). Compared with M group, the number of normal nerve cells in H, R, and U groups increased, with U group having the greatest number (P<0.05). Conclusion: Hyperbaric oxygen and treadmill exercise can improve the endoplasmic reticulum stress and the development of neuronal dendritic spine, inhibit the activation of JAK/STAT signaling pathway, and the combined effect is better. |
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