摘要
目的观察电针对缺血性脑卒中大鼠神经行为学及脑组织结构的影响,探讨电针通过调控NOD样受体热蛋白结构域相关蛋白3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)炎性小体途径改善缺血性脑卒中大鼠神经功能的机制。方法36只健康雄性SD大鼠采用随机数字表随机分为假手术组12只及手术组24只,手术组大鼠采用Longa等改良线栓法制备大鼠脑缺血再灌注损伤(MCAO)模型。术后2 h行神经行为学评分,造模成功的大鼠再随机分为模型组及电针组。电针组予电针“曲池”“足三里”连续干预7 d,假手术组及模型组不做干预。干预完成后分别评估各组大鼠神经功能缺损情况,HE染色观察脑组织病理学变化,QPCR及Western blot法检测炎性小体相关蛋白NLRP3、ASC、Caspase-1的表达水平,ELISA检测各组大鼠血清IL-1β和IL-18炎性因子表达。结果与假手术组相比,模型组大鼠神经功能评分均显著提高,差异有高度统计学意义(P<0.01),脑组织缺血皮质区神经元胞体缩小变形,核固缩明显,大鼠脑组织缺血周围区炎性因子相关蛋白NLRP3、ASC、Caspase-1的mRNA及蛋白表达水平升高,差异有高度统计学意义(P<0.01),血清IL-1β和IL-18炎症因子表达增加,差异有高度统计学意义(P<0.01);经电针干预7 d后,电针组神经评分功能较模型组显著下降,差异有统计学意义(P<0.05),所见的病理损伤减少;炎性小体相关蛋白NLRP3、ASC、Caspase-1的表达下降,差异有统计学意义(P<0.05),且电针下调了血清IL-1β和IL-18的表达,差异有统计学意义(P<0.05)。结论电针“曲池”“足三里”可减轻缺血性脑卒中大鼠神经功能缺损症状,减少脑组织病理损伤,下调NLRP3、ASC、Caspase-1表达水平,其机制可能与调控NLRP3炎性小体途径抗细胞焦亡,减轻脑缺血再灌注损伤有关。
Objective To observe the effects of electroacupuncture on neurobehavior and brain tissue structure in ischemic stroke rats,and to investigate the mechanism of electroacupuncture against cerebral ischemia-reperfusion(I/R)injury by inhibiting the NOD-like receptor hot protein domain-associated protein 3(NLRP3)pathway.Methods A total of 36 healthy male Sprague-Dawley(SD)rats were randomly divided into sham group(n=12)and surgery group(n=24).Rats in the surgery group were subjected to the modeling of middle cerebral artery occlusion(MCAO)via the modified Longa methods.Model in rats was built to simulate I/R injury.Two hours after cerebral I/R injury,neurological defect scores were graded.Rats with the successful modeling of MCAO were further randomly divided into model group and electroacupuncture group.A 7-day consecutive intervention of electroacupuncture at the Quchi(LI11)and Zusanli(ST36)acupoints was performed only in rats of electroacupuncture group.Neurological functions of the rats were assessed at the end of electroacupuncture.Pathological changes in rat brain were observed by hematoxylin and eosin(H&E)staining.Expression levels of inflammasome-associated proteins(NLRP3,apoptosis-associated speck-like protein[ASC]and Caspase-1)were detected by Western blotting and quantitative polymerase chain reaction(q-PCR).Serum levels of inflammatory factors(interleukin-18[IL-18]and interleukin-1beta[IL-1β])were measured by enzyme-linked immunosorbent assay(ELISA).Results Compared with those of sham group,rats in the model group presented significantly higher neurological function scores,reduction and deformation of neuronal cell bodies in ischemic cortical areas,nuclear pyknosis,higher mRNA and protein levels of NLRP3,ASC and Caspase-1 in the ischemic peripheral area,and higher serum levels of IL-1βand IL-18(all P<0.01).A 7-day intervention of electroacupuncture significantly reduced neurological function scores,protected pathological changes in the brain,decreased mRNA and protein levels of NLRP3,ASC and Caspase-1 in the ischemic peripheral area,and lowered serum levels of IL-1βand IL-18(all P<0.05).Conclusion Electroacupuncture at the Quchi(LI11)and Zusanli(ST36)acupoints can reduce the neurological deficit,and downregulate NLRP3,ASC and Caspase-1 by regulating the NLRP3 inflammasome pathway and inhibiting cerebral ischemia and reperfusion injury.
作者
陈阿贞
兰岚
谢小文
林志城
CHEN A-zhen;LAN Lan;XIE Xiao-wen;LIN Zhi-cheng
出处
《中国疗养医学》
2024年第5期1-6,共6页
Chinese Journal of Convalescent Medicine
基金
福建省科技厅自然科学基金项目(2020J01759)
福建省中医药科研项目计划(2021zyjc20)
福建省卫健委青年科研课题(2020QNA065)。
关键词
电针
脑缺血再灌注损伤
NOD样受体热蛋白结构域相关蛋白3
炎性小体
细胞焦亡
Electroacupuncture
Cerebral ischemia/reperfusion injury
NOD-like receptor hot protein domain-associated protein 3
Inflammatory bodies
Cell pyroptosis
