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辜克雨, 刘波, 邢冠君, 何鑫, 丁小连.基于cGAS/STING信号通路探讨针康法对HIBD新生大鼠神经炎症的影响[J].湖南中医药大学学报,2025,45(8):1453-1460[点击复制] |
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| 基于cGAS/STING信号通路探讨针康法对HIBD新生大鼠神经炎症的影响 |
| 辜克雨,刘波,邢冠君,何鑫,丁小连 |
| (黑龙江中医药大学, 黑龙江 哈尔滨 150040;黑龙江中医药大学附属第二医院, 黑龙江 哈尔滨 150001) |
| 摘要: |
| 目的 探讨针康法调节环鸟苷酸腺苷酸合成酶（cGAS）/干扰素基因刺激因子（STING）通路相关蛋白表达对缺血缺氧性脑损伤（HIBD）新生大鼠神经炎症的影响。方法 将Wistar大鼠随机分为假手术组、模型组、针刺组、康复组、针康组（n=30）。采用改良Rice-Vannucci法构建HIBD新生大鼠模型，假手术组和模型组不予干预；针刺组进行头穴丛刺治疗，选取百会穴及双侧旁开2 mm处针刺，快速捻转1 min后留针2 h,1次/d；康复组进行转笼转棒训练，1次/d,10 min/次；针康组在头穴丛刺留针期间同步进行转笼转棒训练。干预结束7、14 d后，采用Longa评分法评估大鼠神经功能缺损情况；HE染色观察大鼠缺血侧大脑皮质组织病理学改变；TUNEL法观察神经细胞凋亡率；ELISA测定脑组织肿瘤坏死因子-α(TNF-α)、白细胞介素（IL）-6、IL-1β、IL-18水平；Western blot测定大鼠缺血侧大脑皮质中cGAS、STING、NOD样受体蛋白3(NLRP3)蛋白表达水平。结果 与假手术组比较，模型组7、14 d的Longa评分增加（P<0.01），缺血侧大脑皮质组织细胞萎缩、排列松散、伴胞核模糊及水肿，神经细胞凋亡率升高（P<0.01），脑组织TNF-α、IL-6、IL-1β、IL-18含量和cGAS、STING、NLRP3蛋白相对表达水平升高（P<0.01）。与模型组比较，各治疗组7、14 d的Longa评分降低（P<0.05），大脑皮质病理损伤得到改善，神经细胞凋亡率降低（P<0.05），脑组织TNF-α、IL-6、IL-1β、IL-18含量和cGAS、STING、NLRP3蛋白相对表达水平降低（P<0.05）。与针刺组、康复组比较，针康组7、14 d的Longa评分降低（P<0.05），大脑皮质病理损伤改善更明显，神经细胞凋亡率降低（P<0.05），脑组织TNF-α、IL-6、IL-1β、IL-18含量和cGAS、STING、NLRP3蛋白相对表达水平降低（P<0.05）。结论 针康法可能通过抑制c GAS/STING通路的异常激活，降低NLRP3炎症小体活性，进而改善HIBD新生大鼠神经炎症损伤情况，促进神经功能恢复。 |
| 关键词: 缺血缺氧性脑损伤 针康法 神经炎症 环鸟苷酸腺苷酸合成酶 干扰素基因刺激因子 NLRP3炎症小体 |
| DOI：10.3969/j.issn.1674-070X.2025.08.008 |
| 投稿时间：2025-06-08 |
| 基金项目:黑龙江省卫生健康委员会科研课题(20230303100276);黑龙江中医药大学附属第二医院孙申田青年人才基金项目(2020KY-14) |
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| Effects of acupuncture-rehabilitation therapy on neuroinflammation in neonatal rats with hypoxic-ischemic brain damage based on the c GAS/STING signaling pathway |
| GU Keyu, LIU Bo, XING Guanjun, HE Xin, DING Xiaolian |
| (Heilongjiang University of Chinese Medicine, Harbin, Heilongjiang 150040, China;The Second Hospital of Heilongjiang University of Chinese Medicine, Harbin, Heilongjiang 150001, China) |
| Abstract: |
| Objective To investigate the effects of acupuncture-rehabilitation therapy on neuroinflammation in neonatal rats with hypoxic-ischemic brain damage（HIBD） by regulating the cyclic GMP-AMP synthase（cGAS）/stimulator of interferon genes（STING） pathway-related protein expressions. Methods Wistar rats were randomly divided into sham-operated group, model group, acupuncture group, rehabilitation group, and acupuncture-rehabilitation group（n =30）. The modified Rice-Vannucci method was applied to establish HIBD neonatal rat model. The sham-operated and model groups were given no intervention;the acupuncture group received cluster acupuncture at the scalp acupoints of Baihui（GV20） and two adjacent points located 2 mm lateral to it on both sides, with rapid needle twirling for 1 minute followed by 2-hour needle retention, once per day; the rehabilitation group received rotating cage and rod training, 10 minutes per session, once per day; and the acupuncturerehabilitation group received simultaneous rotating cage and rod training during the needle retention period of scalp cluster acupuncture. On days 7 and 14 after intervention, the neurological deficits of rats were assessed by Longa score; HE staining was used to observe the histopathological changes in the ischemic cerebral cortex of rats; TUNEL assay was used to observe the apoptosis rate of neuronal cells; ELISA was employed to measure the levels of TNF-α, IL-6, IL-1β, and IL-18 in the brain tissues; Western blot was conducted to check the protein expression levels of cGAS, STING, and NLRP3 in the ischemic cortex. Results Compared with the sham-operated group, the model group showed significantly increased Longa scores on days 7 and 14（P＜0.01）, with neuronal atrophy, disorganized cellular arrangement, nuclear blurring, and edema in the ischemic cerebral cortex. The neuronal apoptosis rate increased（P＜0.01）. The levels of TNF-α, IL-6, IL-1β, IL-18, and the relative protein expressions of c GAS, STING, and NLRP3 were all significantly elevated（P＜0.01）. Compared with the model group, all treatment groups showed reduced Longa scores on days 7 and 14（P＜0.05）, improved pathological damage in the cerebral cortex,decreased neuronal apoptosis（P＜0.05）, and reduced content of TNF-α, IL-6, IL-1β, and IL-18, as well as lower relative protein expression levels of cGAS, STING, NLRP3 in brain tissues（P＜0.05）. Compared with the acupuncture and rehabilitation groups,the acupuncture-rehabilitation group demonstrated greater reductions in Longa scores on days 7 and 14（P＜0.05）, more pronounced improvement in pathological damage in the cerebral cortex, decreased neuronal apoptosis（P＜0.05）, and reduced content of TNF-α, IL-6, IL-1β, and IL-18, as well as lower relative protein expression levels of cGAS, STING, and NLRP3 in brain tissues（P＜0.05）. Conclusion Acupuncture-rehabilitation therapy may alleviate neuroinflammatory damage and promote neurological recovery in neonatal rats with HIBD by inhibiting abnormal activation of the cGAS/STING pathway and suppressing NLRP3 inflammasome activity. |
| Key words: hypoxic-ischemic brain damage acupuncture-rehabilitation therapy neuroinflammation cyclic GMP-AMP synthase stimulator of interferon genes NLRP3 inflammasome |
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