针刺研究

2021, v.46(09) 742-750

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蒙医三根平衡针法对抑郁大鼠p11/组织型纤溶酶原激活物/脑源性神经营养因子通路和miRNA-16的影响
Mongolian medicine three acupoints balance needling improves depressive behavior by regulating p11/tPA/BDNF pathway and miRNA-16 of hippocampus and middle raphe nucleus in depression model rats

赛音朝克图;艾丽雅;宋美丽;罗彤;赵俊;阿茹娜;邓套图格;
SAIYIN Chao-ke-tu;AI Li-ya;SONG Mei-li;LUO Tong;ZHAO Jun;ARU Na;DENG Tao-tu-ge;International Mongolian Medical Hospital of Inner Mongolia Autonomous Region;Mongolian Medical College of Inner Mongolia Medical University;School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine;

摘要(Abstract):

目的:观察蒙医三根平衡针法对慢性应激抑郁模型大鼠海马和中缝核区p11/组织型纤溶酶原激活物(tPA)/脑源性神经营养因子(BDNF)通路及miRNA-16的影响,探讨蒙医三根平衡针治疗抑郁的作用机制。方法:SD大鼠随机分为正常组、模型组、药物组和蒙医针组,每组12只,采用慢性温和不可预见性轻度应激法建立抑郁大鼠模型。药物组大鼠给予盐酸氟西汀灌胃,1次/d,连续28 d。蒙医针组大鼠于应激刺激后1 h给予蒙医三根平衡针法干预,1次/d,连续28 d。观察造模前、干预后各组大鼠行为学改变,免疫荧光法检测大鼠中缝核区p11和tPA阳性表达,Western blot法检测大鼠海马和中缝核区p11和tPA表达量,荧光定量PCR法检测大鼠海马和中缝核区miRNA-16和BDNF mRNA表达。结果:干预后与正常组比较,模型组大鼠水平运动次数、垂直运动次数、糖水消耗量、中缝核区tPA和p11阳性表达、海马和中缝核区tPA和p11相对表达量均减少(P<0.05),海马和中缝核区miRNA-16表达增加(P<0.05),而BDNF mRNA表达减少(P<0.05)。与模型组比较,药物组、蒙医针组大鼠水平运动次数、垂直运动次数、糖水消耗量、中缝核区tPA和p11阳性表达、海马和中缝核区tPA和p11相对表达量均增加(P<0.05),药物组大鼠海马miRNA-16、蒙医针组大鼠海马和中缝核区miRNA-16表达均降低(P<0.05),药物组和蒙医针组大鼠海马和中缝核区BDNF mRNA表达均增加(P<0.05)。结论:抑郁模型大鼠海马和中缝核区内miRNA-16能靶向调控BDNF表达,p11/tPA/BDNF信号通路可能参与了蒙医三根平衡针的抗抑郁效应。
Objective To observe the effect of Mongolian medicine three-acupoints balance needling on the expression of p11/tPA/BDNF pathway and miRNA-16 in the hippocampus and middle raphe nucleus(MRN) in chronic stress depression model rats, so as to explore its mechanisms underlying improvement of depression.Methods Male SD rats were randomly divided into blank control, model, medication and Mongolian medicine acupuncture(acupuncture) groups, with 12 rats in each group. The depression model was established by using chronic unpredictable mild stress method. The rats in the medication group received gavage of prozac(2 mg/kg, diluted with normal saline, 1 mg/mL) 1 h after stress stimulation, once per day for 28 days, and those in the acupuncture group received three-acupoints(Heyi, Badagan and Xin) needling, once a day for 28 days. The behavioral changes were detected by using open field test and sugar consumption test before modeling and after the intervention. The immunoactivity of p11 and tPA proteins in the MRN, and their expression levels in both the MRN and hippocampus were detected by using immunofluorescence histochemistry and Western blot, separately, and the expression levels of miRNA-16 and BDNF mRNA in the hippocampus and MRN detected by using real-time quantitative PCR.Results After modeling, the crossing and rearing scores of open field tests and the relative consumption of sucrose in the model group were apparently lower than those of the blank control group(P<0.05), the expression levels of p11 and tPA in the MRN, and those of p11 and tPA proteins and BDNF mRNA in the MRN and hippocampus were significantly down-regulated(P<0.05), while those of miRNA-16 in the hippocampus and MRN were significantly up-regulated(P<0.05). Compared with the model group, the crossing and rearing scores of open field tests and glucose consumption, as well as the expression levels of p11 and tPA proteins and BDNF mRNA in the hippocampus and MRN were obviously increased in both the medication and acupuncture groups(P<0.05), while the expression of miRNA-16 in hippocampus was markedly down-regulated in both the medication and acupuncture groups(P<0.05). No significant differences were found between the acupuncture and medication groups in all the indexes mentioned above(P>0.05). Conclusion Mongolian medicine three-acupoints balance needling can improve the depressive state in depression rats, which may be associated with its effects in up-regulating the expression of p11 and tPA proteins and BDNF mRNA in the hippocampus and MRN and in down-regulating miRNA-16 in the hippocampus, suggesting an involvement of miRNA-16 controlled p11/tPA/BDNF signaling pathway in the antidepressant effect of acupuncture.

关键词(KeyWords): 抑郁症;蒙医三根平衡针法;海马;中缝核;p11/tPA/BDNF通路;miRNA-16
Depression;Mongolian medicine three-acupoints balance needling;Hippocampus;Middle raphe nucleus;p11/tPA/BDNF pathway;miRNA-16

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(No.81760909)

作者(Author): 赛音朝克图;艾丽雅;宋美丽;罗彤;赵俊;阿茹娜;邓套图格;
SAIYIN Chao-ke-tu;AI Li-ya;SONG Mei-li;LUO Tong;ZHAO Jun;ARU Na;DENG Tao-tu-ge;International Mongolian Medical Hospital of Inner Mongolia Autonomous Region;Mongolian Medical College of Inner Mongolia Medical University;School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine;

Email:

DOI: 10.13702/j.1000-0607.200761

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