基于DNA甲基化探讨针刺联合帕罗西汀治疗轻中度抑郁症的疗效机制

Study on the mechanisms of acupuncture combined with paroxetine in the treatment of mild to moderate depression based on DNA methylation analysis

目的:观察针刺结合帕罗西汀对轻中度抑郁症患者的疗效,并基于DNA甲基化探讨其作用机制。方法:符合纳入标准和排除标准的轻中度抑郁症患者,随机分为观察组和对照组(各33例,脱落3例)健康志愿者25例。对照组予以口服帕罗西汀片20 mg/d,共治疗4周;观察组在对照组的基础上给予针灸治疗,选取中脘、气海、足三里、三阴交、水沟、少商、隐白、大陵、上星,留针20 min/次,每周治疗3次,共4周。分别于治疗前、治疗2周、治疗结束、治疗结束2周后(随访),用汉密尔顿抑郁量表-17(HAMD-17)进行疗效评估;健康志愿者仅在入组前评估1次HAMD-17。随机抽取观察组和对照组各25名患者在治疗前和治疗结束后共进行2次外周血检测,健康志愿者仅入组时进行1次,以MassARRAY技术检测患者及健康志愿者脑源性神经营养因子(BDNF)基因启动子Ⅰ甲基化程度,ELISA法检测血清BDNF含量,分析其在两组患者与健康组间的差异,以及两组患者治疗前后及组间的差异。结果:与本组治疗前比较,观察组患者治疗2周、治疗结束及随访时HAMD-17总评分及其中的睡眠障碍因子、焦虑躯体化因子评分均降低(P<0.05),治疗后血清BDNF含量升高(P<0.05);对照组患者治疗2周、治疗结束、随访时HAMD-17总评分及焦虑躯体化因子评分均降低(P<0.05),治疗结束及随访时睡眠障碍因子评分降低(P<0.05),治疗后血清BDNF含量升高(P<0.05)。与同时点对照组比较,观察组治疗2周、治疗结束及随访时的HAMD-17总评分及睡眠障碍因子评分均降低(P<0.05),焦虑躯体化因子评分仅在治疗2周时降低(P<0.05),治疗结束后外周血BDNF CpG31甲基化程度降低(P<0.05),血清BDNF含量升高(P<0.05)。治疗2周后,观察组总有效率为80.00%(24/30),显效率为36.67%(11/30),而对照组总有效率为26.67%(8/30),显效率为0%(0/30),观察组显效率及总有效率均高于对照组(P<0.05);且治疗结束后观察组显效率为70%(21/30),仍高于对照组的40%(12/30,P<0.05)。血清BDNF含量和HAMD-17呈负相关(ρ=-0.686,P<0.01)。结论:针刺结合帕罗西汀治疗轻中度抑郁症的疗效比单纯口服帕罗西汀更有优势,且改善睡眠障碍和焦虑躯体化的症状起效更快。针刺可能是通过降低外周血BDNF CpG31甲基化程度,升高血清BDNF含量而起到抗抑郁作用。

Objective To explore the therapeutic effect of acupuncture combined with paroxetine for mild to moderate depression and the regulatory role of brain derived neurotrophic factor(BDNF)in patients based on DNA methylation.Methods A total of 66 patients with mild to moderate depression who met the inclusion and exclusion criteria were randomly divided into an observation(acupuncture+medication)group and a control(medication)group,with 33 patients in each group,and other 25 healthy volunteers were taken as the healthy group.The patients of the control group were treated by oral administration of paroxetine 20 mg/d for 4 weeks.The patients of the observation group were treated by acupuncture stimulation of Zhongwan(CV12),Qihai(CV6),Zusanli(ST36),Sanyinjiao(SP6),Shangxing(GV23),Shuigou(GV26),Shaoshang(LU11),Yinbai(SP1)and Daling(PC7)(for 20 min,3 times a week for 4 weeks)on the basis of medication treatment(the same as that of the control group).Before treatment,2 and 4 weeks of treatment,and 2 weeks of follow-up,the therapeutic effect was assessed using Hamilton Depression Scale 17(HAMD-17).The SPSS25.0 software was used to form a randomized grouping and to randomly select 25 patients from the observation group and 25 patients from the control group for blood collecting and data analysis.The blood samples were taken for assaying serum BDNF content and the methylation degree of BDNF gene promotor I with ELISA and MassARRAY techniques,respectively.Results 1)In comparison with those before treatment,the total score of HAMD-17,sleep disorder factor score,and anxiety somatization factor score of both the observation and control groups were significantly decreased after 2 and 4 weeks of treatment,and 2 weeks of follow-up(P<0.05),except sleep disorder factor score in the control group after 2 weeks of the treatment.Compared with the same time-points of the control group,the HAMD-17 total score and sleep disorder factor score of the observation group were decreased after 2 and 4 weeks of treatment,and 2 weeks of follow-up(P<0.05),while the anxiety somatization factor score was evidently decreased after 2 weeks of treatment(P<0.05).2)Following 2 weeks of treatment,the total effective rate and markedly effective rate of the observation group were 80%(24/30)and 36.67%(11/30),respectively,being significantly higher than those[(26.67%and 0%)]of the control group.After 4 weeks of treatment,the markedly effective rate of the observation group was 70.00%(21/30),being significantly higher than that 40%(12/30)of the control group(P<0.05),while the total effective rates of the observation and control groups were the same(100%).3)Before the treatment,comparison among the healthy,observation and control groups showed no statistical significance in the methylation degree of each site(CpG1.2,CpG5.6,CpG8.9,CpG26,CpG27,CpG31,and CpG33.34)of BDNF gene promotor I,while after 4 weeks of the treatment,the methylation degree of CpG31 was considerably lower in the observation group than in the control group(P<0.05).4)Before the treatment,the contents of serum BDNF of both observation and control group had no significant difference,but were evidently lower than that of the healthy group(P<0.05).Compared with that before treatment,the serum BDNF contents in both observation and control groups were significantly increased after the treatment(P<0.05),and was significantly higher in the observation group than in the control group(P<0.05).5)The correlation analysis showed a negative correlation between the BDNF protein content and HAMD-17 score(correlation coefficientρ=-0.686,P<0.01).Conclusion Acupuncture may have an antidepressant role by decreasing CpG31 methylation of BDNF and increasing the serum content of BDNF protein in patients with depression.In addition,acupuncture combined with paroxetine has more advantages in treating mild to moderate depression than oral paroxetine alone,and can improve sleep disorders and anxiety somatization symptoms more quickly.