基于JAK/STAT通路探究温阳通络针灸法对缺血缺氧脑瘫幼鼠相关蛋白JAK2、STAT3的影响

Exploring Effect of Warming Yang and Dredging Collaterals Acupuncture on Related Proteins JAK2 and STAT3 in Ischemic Hypoxic Cerebral Palsy Rats Based on JAK/STAT Pathway

目的探讨温阳通络针灸法对缺血缺氧脑瘫幼鼠的JAK2、p-JAK2、STAT 3、p-STAT3蛋白的影响,方法选用SD清洁级新生大鼠,随机分为空白组、假手术组、模型+药物组、模型+针灸组、模型+针灸+药物组,共6组,每组30只;采用2%戊巴比妥钠麻醉,小心剪开颈部皮肤,分离出迷走神经,并且暴露出颈总动脉,选用6-0的丝线双重结扎颈总动脉,并且中间离断颈总动脉,造成永久性失流,接着将含8%氧气的92%氮氧混合气体以1L/min的流速持续通入缺氧箱中,假手术组仅作皮肤切口,不做缺血处理亦不做缺氧处理,于术后第3天开始干预,药物治疗选用神经节苷脂治疗,隔天1次腹腔注射,针灸治疗穴位选取:百合、大椎、至阳,斜刺百会0.5 cm留针25 min,艾灸大椎、至阳25 min,采用Western Blot(WB)方法分析不同分组中JAK2、p-JAK2、STAT3、p-STAT3表达。结果与假手术组比较,模型+药物组、模型+针灸组、模型+药物+针灸组的p-JAK2、p-STAT3的表达均升高,差异有统计学意义(P<0.05),与模型组比较,模型+药物组、模型+针灸组、模型+药物+针灸组的p-JAK2、p-STAT3表达均降低,差异有统计学意义(P<0.05)。结论温阳通络针灸可明显下调缺血缺氧脑瘫幼鼠p-JAK2、p-STAT3的蛋白表达,减少缺血缺氧对脑组织造成的损伤,促进神经功能恢复。

Objective To investigate the effects of warming Yang and dredging collaterals acupuncture on JAK2,p-JAK2,STAT3 and p-STAT3 proteins in ischemic hypoxic cerebral palsy young rats.Methods SD clean neonatal rats were randomly divided into 6 groups,including blank group,sham operation group,model+drug group,model+acupuncture group,and model+acupuncture+drug group,with 30 rats in each group.The neck skin was carefully cut open to separate the vagus nerve and expose the common carotid artery.6-0 silk thread was used to double ligate the common carotid artery,and the common carotid artery was severed in the middle to cause permanent loss of flow.Then 92%nitrogen oxygen mixture containing 8%oxygen was continuously fed into the anoxic chamber at a flow rate of 1 L/min.In the sham operation group,only skin incision was made without ischemia or hypoxia treatment,and the intervention was started on the third day after surgery.Ganglioside was selected for drug treatment,and the intraperitoneal injection was performed every other day.The acupuncture treatment selected Baihui(DU20),Dazhui(DU14)and Zhiyang(DU9).Oblique needling of Baihui(DU20)0.5 cm with needle retention for 25 minutes,moxibustion of Dazhui(DU14)and Zhiyang(DU9)for 25 minutes.The expressions of JAK2,p-JAK2,STAT3 and p-STAT3 in different groups were analyzed by Western Blot(WB).Results Compared with those of the sham operation group,the expressions of P-JAK2 and P-STAT3 in model+drug group,model+acupuncture group and model+drug+acupuncture group were increased(P<0.05),while the expressions of P-JAK2 and P-STAT3 in model+drug group,model+acupuncture group and model+drug+acupuncture group were decreased(P<0.05).Conclusion Warming Yang and dredging collaterals acupuncture can significantly down-regulate the protein expressions of p-JAK2 and p-STAT3 in ischemic hypoxic cerebral palsy young mice,reduce the damage caused by ischemia and hypoxia to brain tissue,and promote the recovery of nerve function.