电针促进大鼠脊髓缺血再灌注损伤后内源性神经干细胞增殖

Electroacupuncture promotes the proliferation of endogenous neural stem cells after spinal cord ischemia-reperfusion injury in rats

目的探讨电针(electroacupuncture,EA)对大鼠脊髓缺血再灌注损伤(spinalcord ischemia/reperfusion injury,SCIRI)后内源性神经干细胞(endogenous neural stem cells,eNSCs)的保护作用及分子机制。方法成年雄性SD大鼠40只,按照随机数字表法分为五组:假手术组(Sham)、缺血再灌注组(IR)、缺血再灌注+XAV939组(IR+XAV939)、缺血再灌注+电针刺激组(IR+EA)、缺血再灌注+电针刺激+XAV939组(IR+EA+XAV939)。造模成功后,IR+XAV939组和IR+EA+XAV939组在损伤后30min立即腹腔注射Wnt/β-catenin信号通路抑制剂XAV939(20mg/kg),其余组注射等容量2%DMSO。IR+EA组和IR+EA+XAV939在损伤后第二天给予电针刺激(30min/次),连续5天。术后第7天,Tarlov评分评价大鼠后肢运动功能,免疫荧光检测eNSCs标记蛋白巢蛋白(nestin)表达,Western blot检测β-连环蛋白(β-catenin)、nestin蛋白表达。结果 SCIRI后,脊髓eNSCs增加,nestin和β-catenin蛋白表达增加;EA刺激后,eNSCs增生更明显,nestin和β-catenin蛋白表达增多,EA对缺血后的脊髓具有保护作用;XAV939抑制了β-catenin蛋白表达,同时抑制了eNSCs增殖,逆转了EA的保护作用。结论 EA通过激活Wnt/β-catenin通路促进大鼠SCIRI后eNSCs的增殖。

Objective To explore the protective effects and molecules mechanisms of electroacupuncture(EA) on endogenous neural stem cells(eNSCs) after spinal cord ischemiareperfusion injury(SCIRI) in rats. Methods 40 adult male SD rats were divided into five groups according to the random number table: sham operation group(Sham),ischemiareperfusion group(IR), ischemia-reperfusion+XAV939 group(IR+XAV939), ischemiareperfusion+electroacupuncture stimulation(IR+EA), ischemia reperfusion + electroacupuncture stimulation + XAV939(IR+EA+XAV939). After the SCIRI model was successfully established, the IR+XAV939 group and IR+EA+XAV939 group were intraperitoneally injected with Wnt/β-catenin signaling pathway inhibitor XAV939(20 mg/kg) immediately after 30 minutes of SCIRI. The other groups were injected with equal volume of 2% DMSO, IR+EA group and IR+EA+XAV939 were given electroacupuncture stimulation(30 min/time) on the second day after spinal cord ischemia for 5 consecutive days. On the seventh day after the operation,Tarlov score was used to score the motor function of the hind limbs of the rats. The expression of the eNSCs marker protein nestinafter SCIRI was detected by immunofluorescence, and the expression of β-catenin and nestin protein was detected by Western blot. Results After SCIRI, spinal cord eNSCs was increased, nestin and β-catenin protein expression increased;was eNSCs proliferation was more obvious after electroacupuncture stimulation, nestin and β-catenin protein expression was increased. EA has a protective effect on the spinal cord after ischemia. XAV939 inhibited β-catenin protein expression, while inhibiting eNSCs proliferation, and reversed the protective effect of electroacupuncture. Conclusion Electroacupuncture promotes the proliferation of eNSCs after SCIRI in rats by activating Wnt/β-catenin pathway.