基于Nrf2/ARE信号通路探究“输合配穴”点穴法对HIBD仔鼠Keap1、γ-GCS表达影响

Effect Exploration of the Keap1,γ-GCS on“Shu and He Acupoints”Acupuncture Method on HIBD Premature Rats Based on Nrf2/ARE Signal Pathway

目的基于核因子E2相关因子2(Nrf2)/抗氧化反应元件(ARE)信号通路观察“输合配穴-抑木扶土”点穴法治疗新生儿缺氧缺血性脑损伤(HIBD)仔鼠的作用机制。方法将SPF级新生3日龄SD仔鼠30只通过随机数字表法分成空白组、模型组、点穴组,每组10只。通过改良Rice-Vannuncci法建立仔鼠HIBD模型,对点穴组模型选取手阳明经和足阳明经的输穴和合穴进行治疗。分别在第0、7天进行神经行为学检测,取脑组织标本观察病理学结果;采用Western blot、RT-PCR检测脑组织样品Nrf2/ARE信号通路中Nrf2、Keap1及γ-GCS的表达量。结果点穴组脑组织中Nrf2的mRNA及蛋白与γ-GCS的mRNA及蛋白表达较模型组明显升高;Keap1的mRNA及蛋白则低于模型组。结论“输合配穴-抑木扶土”点穴法可以通过Nrf2/ARE信号通路调控抗氧化反应治疗HIBD。

Objective Based on the nuclear factor E2-related factor 2(Nrf2)/antioxidant response element(ARE)signaling pathway,to observe acupuncture method of“Shu and He acupoints-inhibiting wood and strengthening earth”in the treatment of HIBD premature rats.Methods 30 SPF 3-day-old Sprague-Dawley(SD)premature rats were divided into blank control group,model group and acupuncture group(n=10)randomly.The HIBD model of the premature rats was established by the improved RiceVannuncci method;the“Shu and He”acupoints of the large intestine channel of hand Yangming and the stomach channel of foot Yangming were selected for the acupuncture group model.Neurobehavioral tests were performed on the 0th and 7th days,and brain tissue samples were taken to observe the pathological results;Western blot and RT-PCR were used to detect the expression of Nrf2,Keap1 andγ-GCS in the Nrf2/ARE signal pathway of brain tissue samples.Results The expressions of Nrf2 mRNA and protein andγ-GCS mRNA and protein in the brain tissue of the acupuncture group were significantly higher than those of the model group;the mRNA and protein of Keap1 were lower than those of the model group.Conclusion The acupuncture method of“Shu and He acupoints-inhibiting wood and strengthening earth”can regulate the antioxidant response to treat HIBD through the Nrf2/ARE signal pathway.