摘要
目的:探讨电针干预对创伤后应激障碍(PTSD)大鼠焦虑样行为的改善作用,以及其通过糖酵解/组蛋白H4赖氨酸12乳酸化(H4K12la)/缺氧诱导因子1α(HIF-1α)环路发挥调控作用的分子机制。方法:雄性SD大鼠按随机数字表法分为空白组、模型组和电针组,每组10只。用单一持续应激法建立PTSD大鼠模型。电针组针刺“百会”“神庭”及电针双侧“肝俞”“肾俞”,每次20 min,每日1次,共3周。采用旷场实验和高架十字迷宫实验评估大鼠焦虑样行为;采用HE染色观察海马组织形态变化;试剂盒检测海马糖酵解代谢产物乳酸、葡萄糖、三磷酸腺苷(ATP)、丙酮酸含量;荧光定量PCR法及Western blot法检测海马H4K12la、HIF-1α及糖酵解途径关键酶乳酸脱氢酶A(LDHA)和丙酮酸激酶M2(PKM2)的mRNA和蛋白相对表达水平;免疫组织化学法检测H4K12la与HIF-1α在海马区的阳性表达。结果:与空白组相比,模型组大鼠开臂进入次数百分比及开臂停留时间百分比降低(P<0.01),移动总距离、中央区域移动距离下降(P<0.01);海马病理结构紊乱,糖酵解代谢物积累,ATP含量降低(P<0.01),葡萄糖、乳酸含量升高(P<0.01),PKM2、LDHA的mRNA相对表达水平升高(P<0.01,P<0.05),H4K12la、HIF-1α、PKM2、LDHA蛋白表达水平升高(P<0.01),HIF-1α和H4K12la阳性表达升高(P<0.01)。与模型组比较,电针组大鼠开臂进入次数百分比及开臂停留时间百分比升高(P<0.01),移动总距离和中央区域移动距离上升(P<0.01);海马多数锥体细胞形态规则,糖酵解代谢物中ATP含量上升(P<0.01),葡萄糖和乳酸含量降低(P<0.01),PKM2和LDHA的mRNA表达水平降低(P<0.05),H4K12la、HIF-1α、PKM2、LDHA蛋白表达降低(P<0.01),HIF-1α和H4K12la阳性表达面积百分比降低(P<0.01)。结论:电针可通过调控糖酵解/H4K12la/HIF-1α信号环路,改善海马区炎性反应,从而缓解PTSD大鼠的焦虑样行为。
Objective To observe the effect of electroacupuncture(EA)on anxiety-like behavior and activity of glycolysis/histone H4 lysine 12 lactylation(H4K12la)/hypoxia-inducible factor 1α(HIF-1α)signaling in rats with posttraumatic stress disorder(PTSD),so as to explore its underlying molecular mechanisms in the improvement of PTSDinduced anxiety-like behavior.Methods Male SD rats were randomly divided into control,PTSD model,and EA groups,with 10 rats in each group.The PTSD model was established by using a modified single prolonged stress and shock(SPS&S)protocol.The rats in the EA group received EA stimulation(2 Hz/100 Hz,1 mA)at bilateral“Ganshu”(BL18)and“Shenshu”(BL23),and acupuncture at“Baihui”(GV20),“Shenting”(GV24).The anxiety-like behavior was assessed by using both open field(OF)test and elevated plus maze(EPM)test,separately.Morphological changes of hippocampal neurons were observed by HE staining.Glycolysis-related metabolites including lactate,glucose,ATP and pyruvate were measured using biochemical kits.The expressions of H4K12la,HIF-1α,and key glycolytic enzymes[lactate dehydrogenase(LDHA)and pyruvate kinases type M2(PKM2)]in the hippocampal tissue were evaluated by qPCR and Western blot,separately.The spatial distribution of H4K12la and HIF-1αin the hippocampus was further analyzed by immunohistochemistry(IHC).Results The anxiety-like behavior tests showed that the percentages of entries and time spent in the open arms of the EPM test,the total distance and the distance travelled in the center zone of OF test were apparently lower in the model group than in the control group(P<0.01),and significantly higher in the EA group than in the model group(P<0.01).In contrast to the control group,the model group displayed a decrease in the ATP content(P<0.01),and an increase in the contents of glucose,lactic acid,and the expression levels of PKM2 and LDHA mRNAs and proteins,and H4K12la and HIF-1αproteins and immunoactivity in the hippocampal tissues(P<0.05,P<0.01).After EA intervention,the decreased level of ATP and the increased levels of glucose,lactic acid,and expression levels of PKM2 and LDHA mRNAs and proteins,and H4K12la and HIF-1αproteins and immunoactivity were reversed(P<0.05,P<0.01).No significant changes were found in the levels of pyruvate,and expression of H4K12la and HIF-1αmRNAs after modeling and after EA intervention.H.E.staining showed that many hippocampal pyramidal cells were shriveled,with irregular cell shapes,reduced volume and a small number of tiny vacuoles in the model group,while a large number of pyramidal cells were found to have regular shapes,neat and dense arrangement,and a few of shrunken pyramidal cells in the EA group.Conclusion EA intervention can improve the anxiety-like behavior in PTSD rats,which may be related to its functions in down-regulating the glycolysis/H4K12la/HIF-1αpathway and mitigating inflammation in the hippocampus.
作者
程洁
马丹丹
刘广
宋凯
张虹
CHENG Jie;MA Dan-dan;LIU Guang;SONG Kai;ZHANG Hong(Chengdu University of Traditional Chinese Medicine,Chengdu 610000,China;Sichuan Vocational College of Health and Rehabilitation,Zigong 643000,Sichuan Province)
出处
《针刺研究》
北大核心
2026年第1期106-114,共9页
Acupuncture Research
基金
国家中医药管理局专项经费项目(No.500006007)
四川省自然科学基金项目(No.2023NSFSC1821)。
关键词
电针
创伤后应激障碍
焦虑样行为
糖酵解
组蛋白乳酸化
Electroacupuncture
Post-traumatic stress disorder
Anxiety-like behavior
Glycolysis
Histone lactylation
