通督启神电针法对慢性不可预知的应激抑郁大鼠下丘脑PKC/NADPH氧化应激途径的影响  

Effects of Tongdu Qishen electroacupuncture on PKC/NADPH oxidative stress pathway in hypothalamus of chronic unpredictable stress depression rats

作　　者：郭卓[1] 赵静[1] 郭倩[1] 徐舒亭 刘进娜[1] Guo Zhuo;Zhao Jing;Guo Qian;Xu Shuting;Liu Jinna(School of Chinese Integrative Medicine,Hebei Medical University,Shijiazhuang 050030,China)

机构地区：[1]河北医科大学中西医结合学院,石家庄050030

出　　处：《国际中医中药杂志》2024年第4期458-464,共7页International Journal of Traditional Chinese Medicine

基　　金：河北省中医药管理局科研计划项目(2023130、2022131);河北医科大学青年科技创新人才托举培育计划项目(TJZR202010)。

摘　　要：目的观察通督启神电针法的抗抑郁作用,探讨其调控抑郁模型大鼠蛋白激酶C(PKC)/还原型辅酶Ⅱ(NADPH)氧化应激途径的作用机制。方法将32只SD大鼠按随机数字表法分为空白对照组、模型组、通督启神电针组和艾司西酞普兰组。模型组、通督启神电针组及艾司西酞普兰组采用慢性不可预知的应激建立抑郁模型。开始造模后,通督启神电针组每日施以电针治疗,15 min/次;艾司西酞普兰组灌胃艾司西酞普兰30 mg/kg。实验开始前1天和实验第28天称重,观察体重增长情况,并进行糖水偏好实验及旷场实验。采用Western blot免染总蛋白归一法检测大鼠下丘脑组织蛋白激酶Cα(PKCα)、NADPH氧化酶活化蛋白p47(p47phox)、Ras相关C3肉毒菌素物底物1(RAC1)蛋白表达,应用免疫荧光技术检测NADPH氧化酶2(NOX2)阳性面积比率;使用DCFH-DA荧光探针技术检测活性氧(ROS)含量。结果与模型组比较,通督启神电针组和艾司西酞普兰组大鼠体重增长量升高(P<0.01),糖水偏好指数升高(P<0.01),旷场实验总移动距离和中心区停留时间延长(P<0.01或P<0.05);下丘脑区PKCα、p47phox、RAC1蛋白表达降低(P<0.05或P<0.01),NOX2蛋白阳性面积比率降低(P<0.05),ROS水平降低(P<0.01)。结论通督启神电针法可改善抑郁大鼠行为方式,减少PKC/NADPH途径的氧化应激反应,减少ROS生成,从而减轻氧化应激损伤,缓解抑郁症状。Objective To observe the antidepressant effect of Tongdu Qishen electroacupuncture method;To explore its mechanism of regulating the oxidative stress pathway of protein kinase C(PKC)/reduced coenzymeⅡ(NADPH)in depression model rats.Methods Totally 32 SD rats were divided into control group,model group,Tongdu Qishen electroacupuncture group and escitalopram group according to random number table method,with 8 rats in each group.The model of depression was established by chronic unpredictable stress except control group.After the start of modeling,Tongdu Qishen electroacupuncture group was treated with electroacupuncture every day,15 min/time/day;escitalopram group was given 30 mg/kg intragastric intervention.1 day before the start of the experiment and the 28th day of the experiment,the growth of body mass was observed,and sugar preference experiment and open field experiment were performed.The protein expression levels of protein kinase Cα(PKCα),p47phox,t and RAS related C3 botulinum toxin substrate 1(Rac1)in hypothalamus were detected by Western blot,and the positive area ratio of NOX2 protein in hypothalamus was detected by immunofluorescence technique;ROS content in hypothalamus was detected using DCFH-DA fluorescent probe technique.Results Compared with the model group,the Tongdu Qishen electroacupuncture group and the escitalopram group showed the body mass growth(P<0.01)and sugar preference index increased(P<0.01),and the moving distance(P<0.05)and residence time(P<0.01)in the central area of the open field experiment were longer;the protein expression levels of hypothalamic PKCα,p47phox and Rac1 decreased(P<0.05 or P<0.01),the positive area ratio of NOX2 protein decreased(P<0.05),and the level of ROS also decreased significantly(P<0.01)in Tongdu Qishen electroacupuncture group and escitalopram group.Conclusion Tongdu Qishen electroacupuncture group can improve the behavior of depressed rats,inhibit the oxidative stress response of PKC/NADPH pathway,and reduce the production of ROS,thereby reducing the

关 键 词：抑郁症 通督启神电针法 下丘脑 NADPH氧化酶 活性氧 总蛋白归一法 大鼠 

分 类 号：R245.97[医药卫生—针灸推拿学]