基于Trx1/TXNIP通路探讨通督调神针法改善缺血再灌注大鼠海马凋亡机制研究

Study on Mechanism of Dredging Du Meridian and Regulating Spirit Acupuncture to Improve Hippocampal Apoptosis in Ischemia reperfusion Rats Based on Trx1/TXNIP Pathway

目的探讨通督调神针法激活脑缺血再灌注大鼠Trx1,抑制海马凋亡作用机制研究。方法取雄性SD大鼠40只,随机分为空白组,假手术组,模型组及电针组,每组10只。模型组采用Zea-Longa线栓法制备缺血性脑卒中模型,待缺血2 h后,退出线栓进行再灌注。假手术组大鼠仅分离颈总动脉后2 h缝合即可,采用神经功能行为学评分判断大鼠是否成模。成模后电针组选取百会、大椎进行治疗,疗程7 d。治疗结束后收集大脑,TTC染色计算脑梗死面积,qPCR检测大鼠Trx1、TXNIP、Caspase3、Bcl2、Bax基因表达,Elisa检测大鼠海马组织ROS表达,Tunel染色检测大鼠海马组织凋亡情况。结果与假手术组相比较,模型组的脑梗死面积及神经功能行为学评分显著增加(P<0.05);qPCR结果显示,Trx1、Bcl2 mRNA表达显著降低,TXNIP、Caspase3、Bax mRNA表达显著增高(P<0.05);ROS含量显著增高(P<0.05),海马组织凋亡程度显著增加。通督调神针法可以显著降低大鼠神经功能行为学评分、大脑梗死面积及海马凋亡程度,增加Trx1、Bcl2 mRNA表达,降低TXNIP、Caspase3、Bax mRNA表达,降低ROS含量。结论通督调神针法可能通过激活Trx1,抑制TXNIP诱导的凋亡反应,从而发挥缺血再灌注损伤之后大脑保护作用。

Objective To investigate the mechanism of activating Trx1 and inhibiting apoptosis in hippocampus of rats with cerebral ischemia/reperfusion by the acupuncture method of“dredging Du meridian and regulating spirit”.Methods Forty male SD rats were randomly divided into blank group,sham operation group,model group and electroacupuncture group with 10 rats in each group.In the model group,the ischemic stroke model was prepared by Zea Longa thread embolism method.After 2 hours of ischemia,the thread embolism was withdrawn for reperfusion.The rats in the sham operation group could only be sutured 2 h after separating the common carotid artery,and the neurobehavioral score was used to determine whether the rats were modeled.After modeling,the electroacupuncture group selected Baihui(DU20)and Dazhui(DU14)for treatment for 7 days.After the treatment,the brain was collected,and the area of cerebral infarction was calculated by TTC staining.The expressions of Trx1,TXNIP,Caspase3,Bcl2 and Bax genes in rats were detected by qPCR.The expression of ROS in rat hippocampus was detected by ELISA,and the apoptosis in rat hippocampus was detected by Tunel staining.Results Compared with those of the sham operation group,the cerebral infarction area and neurobehavioral score in the model group were significantly increased(P<0.05).The results of qPCR showed that the mRNA expressions of Trx1 and Bcl2 decreased significantly,while the mRNA expressions of TXNIP,Caspase3 and Bax increased significantly(P<0.05).ROS content was significantly increased(P<0.05),and the degree of apoptosis in hippocampus was significantly increased.The dredging Du meridian and regulating spirit acupuncture can significantly reduce the neurological behavioral score,cerebral infarction area and hippocampus apoptosis of rats,increase the expressions of Trx1 and Bcl2 mRNA,reduce the expressions of TXNIP,Caspase3 and Bax mRNA and reduce the content of ROS.Conclusion The dredging Du meridian and regulating spirit acupuncture may play a protective role in brain after ischemia-reperfusion injury by activating Trx1 and inhibiting TXNIP induced apoptosis.