王琴,邵彬,涂维,等.智三针疗法对脑缺血小鼠认知功能缺损及海马神经元细胞凋亡的影响[J].中华物理医学与康复杂志,2024,46(10):865-870
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| 智三针疗法对脑缺血小鼠认知功能缺损及海马神经元细胞凋亡的影响 |
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| DOI：10.3760/cma.j.issn.0254-1424.2024.10.001 |
| 中文关键词: 缺血再灌注 针刺疗法 认知功能 胱天氨酸蛋白水解酶3 |
| 英文关键词: Ischemia Acupuncture Recognition Caspase-3 |
| 基金项目:湖北省自然科学基金一般面上项目（2024AFB1061） |
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| 中文摘要: |
| 目的 探讨智三针疗法对脑缺血小鼠认知功能及海马神经元细胞凋亡的影响及其潜在机制。 方法 选取32只8周C57雄性小鼠随机分为对照组、模型组、伪取穴组和智三针组，每组8只，其中模型组、伪取穴组和智三针组小鼠均接受MCAO/R造模，采用Bederson评分系统和TTC（2，3，5-氯化三苯基四氮唑）染色验证造模成功。对照组和模型组术后不予治疗；伪取穴组和智三针组术后24 h开始针刺治疗，智三针组术后给予针刺神庭及双侧本神穴，而伪取穴组则在神庭及双侧本神穴旁开5 mm处给予针刺，针刺用一次性钢针，进针深度3 mm，留针时间30 min，1次/d，疗程为5 d。治疗结束后24 h，所有小鼠采用水迷宫测试小鼠的认知功能。取小鼠缺血侧海马，用TUNEL法检测小鼠海马神经元细胞凋亡，采用Western blot法检测小鼠海马组织中的半胱氨酸天冬氨酸蛋白酶(caspase)-3、裂解的caspase-3蛋白及p38丝裂原活化蛋白激酶（MAPK）蛋白变化，采用免疫沉淀法检测caspase-3蛋白的总体丝氨酸磷酸化水平。 结果 治疗结束后，与模型组和伪取穴组比较，智三针组小鼠水迷宫测试中的逃避潜伏期显著缩短且穿越次数显著增多（P＜0.05），智三针组小鼠海马神经元TUNEL阳性细胞数显著减少（P＜0.05），海马组织蛋白中caspase-3和裂解的caspase-3含量均显著降低（P＜0.05），智三针组小鼠海马组织中的p38-MAPK蛋白量明显增加（P＜0.05），其下游底物caspase-3蛋白磷酸化水平显著增强（P＜0.05）。 结论 智三针治疗可通过增加海马组织中caspase-3蛋白的丝氨酸磷酸化水平及减少caspase-3活化剪切和神经元细胞凋亡，保护脑缺血小鼠的认知记忆功能。 |
| 英文摘要: |
| Objective To explore the effect of Zhisanzhen acupuncture on cognition ability and apoptosis in the hippocampus using a murine model of cerebral ischemia. Methods Thirty-two male C57 mice were randomly divided into a control group, a model group, a sham acupuncture group and a Zhisanzhen group, each of 8. All except the control group received middle cerebral artery occlusion and reperfusion (MCAO/R) surgery. Successful modeling was confirmed using Bederson scoring and 2,3,5-triphenyltetrazolium chloride staining. Beginning 24 hours after successful modeling, the Zhisanzhen group was given 30 minutes of acupuncture at the Shenting and bilateral Benshen points with the injection 3mm deep daily for five days The sham acupuncture group was given acupuncture at 5 mm next to the Shenting and bilateral Benshen points 3mm deep. Twenty-four hours after the last session of acupuncture, the rats′ cognition was tested using a water maze. After the experiment the hippocampus was sampled on the affected side and any apoptosis of hippocampal neurons was detected using the terminal deoxynucleotidyl transferase nick-end labeling (TUNEL) assays. Changes in caspase-3, cleaved caspase-3 protein and p38 mitogen-activated protein kinase (MAPK) protein in the hippocampal tissue were evaluated using western blotting. The total serine phosphorylation level of caspase-3 protein was detected using immunoprecipitation. Results Compared to the model and the sham groups, the average escape latency in the Zhisanzhen group was significantly shorter and the times crossing the platform was significantly greater. There were significantly fewer TUNEL-positive cells in the Zhisanzhen group on average, and less caspase-3 and cleaved caspase-3, but significantly more p38-MAPK and greater serine phosphorylation of its substrate caspase-3. Conclusion Zhisanzhen acupuncture can ameliorate recognition difficulties after cerebral ischemia. It promotes phosphorylation of caspase-3, limiting its cleavage and activation, and decreasing neuron apoptosis in the hippocampus. |
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