基于JAK/STAT通路探讨温阳通络针灸法干预缺氧脑瘫幼鼠的作用机制研究

Study on the Action Mechanism of Wenyang Tongluo Acupuncture for Mice with Hypoxic Cerebral Palsy Based on JAK/STAT Pathway

目的:基于Janus激酶/信号转导与转录激活子(JAK/STAT)通路探讨温阳通络针灸法干预缺氧脑瘫幼鼠的作用机制研究。方法:将40只幼鼠随机分为对照组、模型组、温阳通络针灸组和JAK/STAT通路抑制剂WP1066组各10只。HE染色检测大脑皮质损伤、TUNEL染色检测大脑皮质细胞凋亡、ELISA法检测幼鼠氧化应激指标超氧化物歧化酶(SOD)、丙二醛(MDA)、半胱氨酸天冬氡酸蛋白酶3(Caspase-3)和内源性激活素A(ACT A)水平;Western Blot检测大鼠大脑皮质中JAK2、p-JAK2、STAT3、p-STAT3的表达。结果:与模型组相比,温阳通络针灸组和WP1066组幼鼠平均潜伏时间明显缩短,穿越原平台次数明显增多(P<0.05);HE染色结果显示对照组新生大鼠大脑皮质层未见明显病变、模型组大鼠大脑皮质层损伤最明显、温阳通络针灸组和WP1066组幼鼠大脑皮质层病理损伤减轻;TUNEL检测结果显示,温阳通络针灸组和WP1066组幼鼠模鼠大脑皮质层细胞凋亡率明显低于模型组(P<0.05);ELISA检测结果显示,与模型组相比,温阳通络针灸组和WP1066组幼鼠SOD、ACT A表达明显上升,Caspase-3、MDA水平明显下降(P<0.05);Western blot检测结果显示,与模型组相比,温阳通络针灸组和WP1066组幼鼠脑组织p-JAK2、p-STAT3表达水平明显降低(P<0.05)。结论:温阳通络针灸法可以通过抑制JAK2/STAT3信号,恢复缺氧脑瘫幼鼠神经功能和大脑损伤。

Objective:To explore the action mechanism of Wenyang Tongluo acupuncture for mice with hypoxic cerebral palsy based on Janus tyrosine kinase/signal transducer and activator of transcription(JAK/STAT)pathway.Methods:A total of 40 mice were randomly divided into a controlled group,a model group,a Wenyang Tongluo acupuncture group and a JAK/STAT pathway inhibitor(WP1066)group,10 cases in each group.The cerebral cortex injury was detected by HE staining.The apoptosis of cerebral cortex was detected by TUNEL staining.The levels of oxidative stress indexes[superoxide dismutase(SOD),malondialdehyde(MDA),cysteine aspastic protease 3(Caspase-3),endogenous activin A(ACT A)]were detected by ELISA.The expressions of JAK2,p-JAK2,STAT3 and p-STAT3 in cerebral cortex were detected by Western Blot.Results:Compared with the model group,the average latency was significantly shortened in the Wenyang Tongluo acupuncture group and the WP1066 group,and times of crossing the original platform were significantly increased(P<0.05).The results of HE staining showed that there were no significant lesions of cerebral cortex in the controlled group.The cerebral cortex injury in the model group was the most significant,and the pathological damage in the Wenyang Tongluo acupuncture group and the WP1066 group was relieved.TUNEL results showed that apoptosis rate of cerebral cortex in the Wenyang Tongluo acupuncture group and the WP1066 group was significantly lower than that in the model group(P<0.05).ELISA results showed that expression levels of SOD and ACT A were significantly increased in the Wenyang Tongluo acupuncture group and the WP1066 group compared with the model group,while levels of Caspase-3 and MDA were significantly decreased(P<0.05).Western blot results showed that compared with the model group,expression levels of p-JAK2 and p-STAT3 in brain tissues were significantly decreased in the Wenyang Tongluo acupuncture group and the WP1066 group(P<0.05).Conclusion:Wenyang Tongluo acupuncture can recovery nerve function and brain injury in mice with hypoxic cerebral palsy by inhibiting JAK2/STAT3 signals.