文献类型：期刊文献

中文题名：基于Dnmt1介导的Foxp3甲基化和Treg/Th17平衡探究电头针对脑缺血大鼠缺血皮质区炎症反应的影响

英文题名：Exploration on the Effects of Electro-scalp Acupuncture on Inflammatory Response in Ischemic Cortical Area of Cerebral Ischemic Rats Based on Dnmt1-mediated Foxp3 Methylation and the Treg/Th17 Balance

作者：刘翠[1];杨皓雷[1];魏劢[1];刘倩茹[1];井维尧[1];刘莉梅[2];李兴兰[1];王金海[3];杜小正[1]

第一作者：刘翠

机构：[1]甘肃中医药大学针灸推拿学院,甘肃兰州730000;[2]甘肃中医药大学附属医院,甘肃兰州730030;[3]兰州大学第二医院,甘肃兰州730030

第一机构：甘肃中医药大学针灸推拿学院

年份：2026

卷号：33

期号：3

起止页码：89

中文期刊名：中国中医药信息杂志

外文期刊名：Chinese Journal of Information on Traditional Chinese Medicine

基金：国家自然科学基金(82260962、82460965);甘肃省中医药管理局项目(GZKZ-2024-35);甘肃省科技计划项目重点研发计划(20YF8WA097)。

语种：中文

中文关键词：电头针;缺血性脑卒中;Foxp3甲基化;Treg/Th17;炎症反应;大鼠

外文关键词：electro-scalp acupuncture;ischemic stroke;Foxp3 methylation;Treg/Th17;inflammatory response;rats

摘要：目的基于DNA甲基转移酶1(Dnmt1)介导的叉头框蛋白p3(Foxp3)甲基化和Treg/Th17平衡探究电头针缓解脑缺血大鼠炎症反应的具体机制。方法90只SPF级SD大鼠随机取12只作为假手术组,其余大鼠制备脑缺血模型。将成模大鼠随机分为模型组、地西他滨组、电头针组、亚砷酸钠组、电头针+亚砷酸钠组,每组12只。地西他滨组腹腔注射地西他滨0.4 mg/kg,电头针组于双侧“顶颞前斜线”予电针刺激,频率2 Hz/100 Hz,电流1 mA,30 min/次,亚砷酸钠组尾静脉注射亚砷酸钠溶液0.08 mg/kg,电头针+亚砷酸钠组在注射亚砷酸钠基础上行电头针干预,均1次/d,连续7 d。对大鼠进行神经功能缺损评分,TTC染色检测脑梗死体积,焦磷酸测序检测脾组织Foxp3基因启动子甲基化水平,RT-PCR检测脾组织Foxp3、Dnmt1、RORγt mRNA表达,Western blot检测缺血皮质区Foxp3、RORγt蛋白表达,免疫组化检测缺血皮质区白细胞介素(IL)-6、IL-17、肿瘤坏死因子(TNF)-α、IL-10、转化生长因子(TGF)-β表达,流式细胞术检测缺血皮质区Foxp3+、IL-17A+细胞比例。结果与假手术组比较,模型组大鼠神经功能缺损评分明显升高(P<0.01),脑梗死体积明显增大(P<0.01),脾组织Foxp3基因启动子甲基化水平明显升高(P<0.05),Foxp3 mRNA表达明显降低,Dnmt1、RORγt mRNA表达明显升高(P<0.01),缺血皮质区Foxp3蛋白表达明显降低,RORγt蛋白表达明显升高(P<0.01),IL-6、IL-17、TNF-α阳性表达明显升高,IL-10、TGF-β阳性表达明显降低(P<0.05),Treg/Th17细胞比例明显降低(P<0.01);与模型组比较,电头针组、地西他滨组大鼠神经功能缺损评分明显降低(P<0.01),脑梗死体积明显缩小(P<0.01),脾组织Foxp3基因甲基化水平明显降低(P<0.05),Foxp3 mRNA表达明显升高,Dnmt1、RORγt mRNA表达明显降低(P<0.01),缺血皮质区Foxp3蛋白表达明显升高,RORγt蛋白表达明显降低(P<0.01),IL-6、IL-17、TNF-α阳性表达明显降低,IL-10、TGF-β阳性表达明显升高(P<0.05),Treg/Th17细胞比例明显升高(P<0.01)。结论电头针可通过抑制脑缺血大鼠Dnmt1表达降低Foxp3基因甲基化水平,恢复Treg/Th17细胞平衡,调节炎症因子分泌,减轻脑组织炎症损伤,发挥抗脑缺血作用。
Objective To explore the specific mechanism of electro-scalp acupuncture(ESA)in alleviating the inflammatory response in cerebral ischemic rats based on DNA methyltransferase 1(Dnmt1)-mediated methylation of forkhead box protein p3(Foxp3)and Treg/Th17 balance.Methods A total of 90 SPF-grade SD rats were randomly selected 12 as sham-operation group,and the rest were used to establish cerebral ischemia model.The successfully modeled rats were randomly divided into model group,5-Aza group,ESA group,sodium arsenite(SA)group and ESA+SA group,with 12 rats in each group.The 5-Aza group received intraperitoneal injection of decitabine at a dosage of 0.4 mg/kg.The ESA group received electroacupuncture stimulation on the bilateral anterior parietaltemporal oblique lines with a frequency of 2 Hz/100 Hz,a current of 1 mA,and the needles were retained for 30 minutes.The SA group received an injection of sodium arsenite solution at 1/3 of the tail tip at a dosage of 0.08 mg/kg.The ESA+SA group received electro-scalp acupuncture intervention on the basis of sodium arsenite solution injection.All the above interventions were performed once a day for 7 consecutive days.Neurological deficit scores were evaluated,TTC staining was used to detect cerebral infarct volume,pyrosequencing was used to detect Foxp3 methylation level in spleen tissue,RT-PCR was used to detect the mRNA expressions of Foxp3,Dnmt1 and RORγt in spleen tissue;Western blot was used to detect the protein expressions of Foxp3 and RORγt in ischemic cortical area,immunohistochemistry was used to detect the expression of IL-6,IL-17,TNF-α,IL-10 and TGF-βin ischemic cortical cortical area,flow cytometry was used to detect Foxp3+,IL-17A+cell proportion in ischemic cortical area.Results Compared with the sham-operation group,the neurological deficit score of the model group significantly increased(P<0.01),the volume of cerebral infarction significantly increased(P<0.01),the methylation level of Foxp3 gene promoter in spleen tissue significantly increased(P<0.05),the expression of Foxp3 mRNA significantly decreased,the expressions of Dnmt1 and RORγt mRNA significantly increased(P<0.01),the expression of Foxp3 protein in ischemic cortical area significantly decreased,RORγt protein significantly increased(P<0.01),the positive expressions of IL-6,IL-17 and TNF-α significantly increased,and the positive expressions of IL-10 and TGF-β significantly decreased(P<0.05),the proportion of Treg/Th17 cells significantly decreased(P<0.01).Compared with the model group,the scores of neurological deficits in ESA group and 5-Aza group significantly decreased(P<0.01),the volume of cerebral infarction was significantly reduced(P<0.01),the methylation level of Foxp3 gene in spleen tissue significantly decreased(P<0.05),the expression of Foxp3 mRNA significantly increased,and the expressions of Dnmt1 and RORγt mRNA significantly decreased(P<0.01),the expression of Foxp3 protein in ischemic cortical area significantly increased,the expression of RORγt protein significantly decreased(P<0.01),the positive expressions of IL-6,IL-17 and TNF-α significantly decreased,and the positive expressions of IL-10 and TGF-β significantly increased(P<0.05),the proportion of Treg/Th17 cells significantly increased(P<0.01).Conclusion ESA can reduce the methylation level of Foxp3 gene by inhibiting the expression of Dnmt1 in rats with cerebral ischemia,restore the balance of Treg/Th17 cells,regulate the secretion of inflammatory factors,reduce the inflammatory injury of brain tissue,and play an anti-cerebral ischemia role.