中国组织工程研究

中国组织工程研究 ›› 2024, Vol. 28 ›› Issue (34): 5473-5479.doi: 10.12307/2024.721

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

艾灸抑制NLRP3/Caspase-1通路介导的细胞焦亡减轻脑缺血再灌注损伤

于燕艳1，刘  娟2 ，杨  越1，王千慧1，李  姗1，蒋  洁1   

  1. 1新疆医科大学中医学院，新疆维吾尔自治区乌鲁木齐市   830054；2新疆医科大学第四临床医学院，新疆维吾尔自治区乌鲁木齐市   830054
  • 收稿日期:2023-08-31 接受日期:2023-10-30 出版日期:2024-12-08 发布日期:2024-03-14
  • 通讯作者: 蒋洁，硕士，副教授，新疆医科大学中医学院，新疆维吾尔自治区乌鲁木齐市 830054
  • 作者简介:于燕艳，女，1993年生，黑龙江省饶河县人，汉族，新疆医科大学在读硕士，主要从事针灸治疗脑血管疾病的研究。
  • 基金资助:
    新疆维吾尔自治区高校科研计划项目(XJEDU2021Y025)，项目负责人：蒋洁；自治区“十四五”重点学科(特色学科)中医学

Moxibustion inhibits NLRP3/Caspase-1 pathway mediated cell pyroptosis and alleviates cerebral ischemia/reperfusion injury

Yu Yanyan1, Liu Juan2, Yang Yue1, Wang Qianhui1, Li Shan1, Jiang Jie1   

  1. 1College of Traditional Chinese Medicine, Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China; 2Fourth Clinical Medical College of Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China 
  • Received:2023-08-31 Accepted:2023-10-30 Online:2024-12-08 Published:2024-03-14
  • Contact: Jiang Jie, Master, Associate professor, College of Traditional Chinese Medicine, Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • About author:Yu Yanyan, Master candidate, College of Traditional Chinese Medicine, Xinjiang Medical University, Urumqi 830054, Xinjiang Uygur Autonomous Region, China
  • Supported by:
    Xinjiang Uygur Autonomous Region University Research Project, No. XJEDU2021Y025 (to JJ); Key Disciplines (Characteristic Disciplines) Traditional Chinese Medicine during the “14th Five-Year Plan” Period of Xinjiang Uygur Autonomous Region

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摘要：


文题释义：

脑缺血再灌注损伤：脑缺血恢复血液供应后，脑组织会产生一系列应激反应进一步加重脑组织损伤。
细胞焦亡：又称细胞炎性坏死，是一种程序性细胞死亡，表现为细胞不断膨大直至细胞膜破裂，导致细胞内容物大量释放进而激活强烈的炎症反应。细胞焦亡是机体一种重要的天然免疫反应，在神经系统疾病中发挥重要作用。


背景：研究发现，艾灸能抑制脑缺血/再灌注损伤大鼠血清中的炎症因子，对抗氧化应激，抑制细胞凋亡，有效减轻脑缺血/再灌注损伤。

目的：观察艾灸干预不同时间后脑缺血/再灌注损伤模型大鼠核苷酸结合寡聚化结构域样受体蛋白3(nucleotide-binding oligomerization domain-like receptor protein 3，NLRP3)炎性小体、半胱氨酸天冬氨酸酶1(cysteine aspartase，caspase-1)、凋亡相关斑点样蛋白(ASC)、消皮素D(GSDMD)及白细胞介素1β、白细胞介素18表达的变化，并探讨其作用机制。
方法：SD大鼠随机分为假手术组9只和造模组36只，造模组制备大脑中动脉栓塞大鼠模型。造模成功后将造模组大鼠进一步分为模型组、艾灸10 min组、艾灸15 min组、艾灸30 min组，每组9只。艾灸10 min、15 min和30 min组大鼠分别给予艾灸悬灸“百会、大椎、足三里”穴，均干预1次/d，共干预7 d。采用LONGA法评估大鼠神经功能缺损情况，TTC法观察脑组织梗死情况，苏木精-伊红染色观察脑组织病理改变，ELISA法检测血清中白细胞介素1β、白细胞介素18水平，免疫组织化学法和蛋白免疫印迹法测定脑皮质缺血区组织NLRP3、caspase-1、凋亡相关斑点样蛋白、消皮素D蛋白的表达。

结果与结论：①与假手术组比较，模型组大鼠神经功能缺损评分显著升高(P < 0.01)，与模型组比较，艾灸各组大鼠神经功能缺损评分明显降低(P < 0.01)；②与假手术组比较，模型组大鼠脑梗死体积明显增大(P < 0.01)，与模型组比较，艾灸各组大鼠脑梗死体积明显减少(P < 0.01)，艾灸30 min组脑梗死体积最小(P < 0.05)；③与模型组比较，艾灸各组大鼠血清白细胞介素1β、白细胞介素18水平下降(P < 0.01)，与艾灸10 min组比较，艾灸30 min组血清中的炎症因子水平明显下降(P < 0.05)；④与模型组比较，艾灸各组大鼠皮质缺血区NLRP3、凋亡相关斑点样蛋白、Caspase-1、消皮素D蛋白表达显著降低(P < 0.01)，艾灸30 min组蛋白表达明显低于艾灸10 min组和艾灸15 min组(P < 0.05)；⑤结论：艾灸百会、大椎、足三里可减轻脑缺血/再灌注损伤，其中艾灸30 min疗效最佳，其机制可能与抑制NLRP3/Caspase-1通路介导的细胞焦亡有关。

https://orcid.org/0009-0005-7160-608X (于燕艳) 

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

关键词: 艾灸, 细胞焦亡, 脑缺血再灌注损伤, NLRP3, 炎症小体, Caspase-1

Abstract: BACKGROUND: It was found that moxibustion can inhibit the inflammatory factors in the serum of rats with cerebral ischemia/reperfusion injury, resist oxidative stress, inhibit cell apoptosis, and effectively reduce cerebral ischemia/reperfusion injury.
OBJECTIVE: To observe the effects of different moxibustion intervention time on the expression levels of nucleotide binding oligomerization domain-like protein 3 inflammasome (NLRP3), cysteine aspartase (caspase-1), apoptosis-related speck-like protein, exfoliatin-D protein, interleukin-1β and interleukin-18 in rats with cerebral ischemia/reperfusion injury, and to explore its action mechanism.
METHODS: SD rats were randomly divided into sham operation group (n=9) and operation group (n=36). The model of focal cerebral ischemia/reperfusion injury was established by middle cerebral artery occlusion in the operation group. After successful modeling, the rats in the operation group were further divided into model group, moxibustion 10-minute group, moxibustion 15-minute group and moxibustion 30-minute group, with 9 rats in each group. Rats in the moxibustion 10-minute, 15-minute and 30-minute groups were given moxibustion at “Baihui, Dazhui and Zusanli”, respectively, once a day for a total of 7 days. The neurological deficits of rats were evaluated by LONGA method. The cerebral infarction was observed by 2,3,5-triphenyltetrazolium chloride staining. The pathological changes of brain tissue were observed by hematoxylin-eosin staining. The contents of interleukin-1β and interleukin-18 in serum of rats in each group were detected by ELISA. Immunohistochemistry and western blot assay were used to detect the expression levels of NLRP3, caspase-1, apoptosis-related spot-like protein and gasdermin D in the ischemic cortex of rats in each group.
RESULTS AND CONCLUSION: Compared with the sham operation group, the neurological deficit score of the model group was significantly increased (P < 0.01). Compared with the model group, the neurological deficit score of the moxibustion groups was significantly reduced (P < 0.01). Compared with the sham operation group, the infarct volume of the model group was significantly increased (P < 0.01). Compared with the model group, the infarct volume of the moxibustion groups was significantly reduced (P < 0.01); the infarct volume of the rats was smallest in the moxibustion 30-minute group (P < 0.05). Compared with the model group, the contents of inflammatory factors interleukin-1β and interleukin-18 in the serum of rats in the moxibustion groups were decreased (P < 0.01). Compared with the moxibustion 10-minute group, the contents of inflammatory factors in the serum of rats in the moxibustion 30-minute group were significantly decreased (P < 0.05). Compared with the model group, the expression of NLRP3, apoptosis-related spot-like protein, Caspase-1 and gasdermin D protein in the ischemic cortex of the moxibustion groups was significantly decreased (P < 0.01). Compared with the moxibustion 10-minute and 15-minute groups, the expression of protein in the moxibustion 30-minute group was significantly decreased (P < 0.05). It is concluded that moxibustion at Baihui, Dazhui and Zusanli can reduce cerebral ischemia/reperfusion injury, among which moxibustion for 30 minutes has the best effect, and its mechanism may be related to the inhibition of pyroptosis mediated by NLRP3/Caspase-1 pathway. 

Key words: moxibustion, pyroptosis, cerebral ischemia/reperfusion injury, NLRP3, inflammasome, Caspase-1

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