电针干预对糖尿病术后认知功能障碍小鼠线粒体分裂的影响

doi:10.20047/j.issn1673-7210.2023.36.01

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摘要目的观察电针干预对糖尿病术后认知功能障碍（POCD）小鼠线粒体分裂的影响。方法 39只体重（20±2）g的4～6周龄SPF级C57BL/6雄性小鼠高脂高糖饮食16周后腹腔注射链脲佐菌素，1周后空腹血糖≥11.1 mmol/L，即为2型糖尿病实验动物（无小鼠死亡），随后采用随机数字表法将其分为对照组、模型组、电针组，每组13只。对照组除抓取与固定外不做其他干预；模型组与电针组行胫骨骨折髓内固定术建立POCD模型（无小鼠死亡）；模型组与对照组比较，认知能力下降视为造模成功。电针组术前2周取神庭、百会、内关和足三里行电针干预，每次留针30 min，每周连续5 d。术后1 d进行旷场试验与恐惧条件测试评估小鼠认知功能；苏木精-伊红染色法观察海马CA1区病理情况；尼氏染色法观察神经元数量；化学发光法检测腺苷三磷酸（ATP）含量；流式细胞术检测线粒体膜电位（MMP）；Western blot检测小鼠海马发动蛋白相关蛋白1（Drp1）、p-Drp1 Ser616、p-Drp1 Ser637的蛋白表达水平。结果与对照组比较，模型组场景恐惧测试僵直时间缩短（P＜0.05）；与模型组比较，电针组场景恐惧测试僵直时间增加（P＜0.05）。与对照组比较，模型组CA1区神经元损伤加重、神经元数量减少（P＜0.05）；与模型组比较，电针组CA1区神经元损伤减轻、神经元数量增加（P＜0.05）。与对照组比较，模型组ATP含量减少，MMP降低（P＜0.05）；与模型组比较，电针组ATP含量增加，MMP升高（P＜0.05）。与对照组比较，模型组海马组织p-Drp1 Ser616蛋白表达水平升高，p-Drp1 Ser637蛋白表达水平降低（P＜0.05）；与模型组比较，电针组海马组织p-Drp1 Ser616蛋白表达水平降低，p-Drp1 Ser637蛋白表达水平升高（P＜0.05）。结论电针干预可以提高糖尿病小鼠POCD的学习记忆能力，减轻神经元损伤，其机制可能与减少Drp1介导的线粒体过度分裂，改善海马线粒体功能有关。

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	炎茹玉丁玲玲陈梦洁罗见生宁甲齐周瑞玲于波

关键词 ：术后认知功能障碍, 电针, 线粒体分裂

Abstract：Objective To observe the effect of electro-acupuncture intervention on mitochondrial fission in diabetic mice with postoperative cognitive dysfunction (POCD). Methods Thirty-nine SPF-grade C57BL/6 male mice of body weight(20±2) g aged four to six weeks injected intraperitoneally with streptozotocin after 16 weeks of high-fat and high-sugar diet with fasting blood glucose≥11.1 mmol/L after one week, that is, they were experimental animals with diabetes mellitus type 2 (no mice died). Then they were divided into the control group, the model group, and the electro-acupuncture group by random number table method, 13 mice in each group. The control group did not do any other intervention except grasp and fixation. POCD model was established by tibial fracture intra- medullary fixation between model group and electroacupu- ncture group (no mice death). Compared with the control group, the cognitive ability of the model group decreased and the model was considered successful. In the electro-acupuncture group, shéntíng, bǎihuì, nèiguān, and zúsānlǐ were treated with electro-acupuncture intervention two weeks before surgery, and the needles were retained for 30 min each time for five consecutive days every week. The cognitive function of mice was evaluated by open field test and fear conditioning test one day after operation. Hematoxylin-eosin staining was used to observe the pathology of hippocampal CA1 region. The number of neurons was observed by Nissl staining. The content of adenosine triphosphate (ATP) was detected by chemiluminescence. Mitochondrial membrane potential (MMP) was detected by flow cytometry. Western blot analysis was performed to detect the protein expression levels of dynamin-related protein 1(Drp1), p-Drp1 Ser616, and p-Drp1 Ser637 in the hippocampus of mice. Results Compared with the control group, the freezing time of the context test in the model group was reduced (P＜0.05). Compared with the model group, the freezing time of the context test in the electro-acupuncture group was increased (P＜0.05). Compared with the control group, the damage of neurons in CA1 region of the model group was aggravated and the number of neurons was decreased (P＜0.05). Compared with the model group, the damage of neurons in CA1 region was reduced and the number of neurons was increased in electro-acupuncture group (P＜0.05). Compared with the control group, ATP content and MMP in the model group were decreased (P＜0.05). Compared with the model group, ATP content and MMP in electro-acupuncture group were increased (P＜0.05). Compared with the control group, the expression level of p-Drp1 Ser616 protein in hippocampus of model group was increased, while the expression level of p-Drp1 Ser616 protein was decreased (P＜0.05). Compared with the the model group, the expression level of p-Drp1 Ser616 protein in hippocampus of the electro-acupuncture group was decreased, and the expression level of p-Drp1 Ser616 protein was increased (P＜0.05). Conclusion Electro-acupuncture can improve the learning and memory ability of POCD in diabetic mice and reduce the neuronal damage. The mechanism may be related to the reduction of Drp1-mediated mitochondrial hyperfission and the improvement of hippocampal mitochondrial function.

Key words： Postoperative cognitive dysfunction Electro-acupuncture Mitochondrial fission

基金资助:国家自然科学基金资助项目（82374580）；首都卫生发展科研专项项目（首发2022-2-2232）；首都医科大学附属北京中医医院院级课题暨两院合作课题项目（LYYB202216）。

通讯作者: 丁玲玲（1971.8-），女，博士，主任医师，博士生导师；研究方向：中西医结合围手术期脏器保护。

作者简介: 炎茹玉（1998.1-），女，首都医科大学附属中医药临床医学院2021级麻醉学专业在读硕士研究生；研究方向：中西医结合围手术期脏器保护。

引用本文:

炎茹玉丁玲玲陈梦洁罗见生宁甲齐周瑞玲于波. 电针干预对糖尿病术后认知功能障碍小鼠线粒体分裂的影响[J]. 中国医药导报, 2023, 20(36): 4-9.
YAN Ruyu DING Lingling CHEN Mengjie LUO Jiansheng NING Jiaqi ZHOU Ruiling YU Bo. Effect of electro-acupuncture intervention on mitochondrial fission in diabetic mice with postoperative cognitive dysfunction. 中国医药导报, 2023, 20(36): 4-9.

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https://www.yiyaodaobao.com.cn/CN/10.20047/j.issn1673-7210.2023.36.01 或 https://www.yiyaodaobao.com.cn/CN/Y2023/V20/I36/4

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