电针结合经颅交流电刺激对脑缺血大鼠神经炎症和凋亡相关基因表达的影响

Effects of electroacupuncture combined with transcranial alternating current stimulation on inflammatory-apoptotic genes expression of MCAO rats

AbstractObjective： To establish right middle cerebral artery occlusion（MCAO） model in rats and to investigate the mechanism underlying motor function regulation by transcranial alternating current stimulation（tACS） intervention. Furthermore， to dynamically observe the effects of electroacupuncture combined with tACS on neurological deficit scores(NDS)， cerebral blood flow， inflammatory-cell apoptosis gene of MCAO model rats and to explore the mechanism of cerebral and neural regulation on motor function rehabilitation after cerebral ischemia reperfusion.Method： Forty SD rats were randomly divided into sham-operation group （S group）， model group （M group）， electroacupuncture group （EA group）， transcranial alternating current stimulation group （T group） and electroacupuncture combined with transcranial alternating current stimulation group （EA+T group）. After 2 h ischemia-reperfusion， EA group was given bilateral Qu chi （LI 11） and Zu san li （ST 36）electroacupuncture under anesthesia. Right M1 was selected for tACS in T group. EA+T group was treated with EA and tACS together. S and M group were treated with anesthesia for 30min per time， for 7 days. The data from before modling（B） to after modling（D7） were recorded, including neurological deficit score and blood flow of the right middle cerebral artery by laser doppler flowmetry. RT-PCR was used to analyze inflammation-cell apoptosis gene expression at D7.Result：Neurological deficit score: at 2h， D1， M group， EA group， T group and EA+T group increased significantly compared with other time（P<0.05）. At D3， D5， D7， S group decreased significantly compared with other time（P<0.05）, while M group increased significantly compared with other group（P<0.05）. EA group， T group and EA+T group were significantly different in all times（P<0.05）. At 2h， D1， D3， D5， D7, M group increased significantly compared with B. At D1， D3， D5， D7, NDS decreased significantly compared with that at 2h（P<0.05）. Blood flow： EA+T group and S group decreased at 2h， increased at D1 and decreased at D3. EA group increased at D3. However, M group decreased significantly compared with other group at D1， D3 and D5（P<0.05）. RT-PCR： motor cortex ΔCt analysis： Caspase 12 in EA+T group decreased significantly compared with T group（P<0.05）， IL-1β and NLRP1a in EA group and T group decreased significantly compared with those in S group（P<0.05）. 2-ΔΔCt analysis in ischemic region： ATF4 in M group increased significantly compared with that in other groups （P<0.05）； Bcl 2 in M group increased significantly compared with that in S group， EA group and EA+T group（P<0.05）； Bax， Caspase 12， C-fos in M group increased significantly compared with those in S Group（P<0.05）.Conclusion： Electroacupuncture combined with tACS can modulate the inflammatory response and inhibit cell apoptosis through regulating ATF4、Bcl-2、Bax、Caspase 12、C-fos and can be a new strategy for the treatment of ischemic stroke.