不同经穴组合针刺对失眠大鼠松果体褪黑素含量的影响

Manual Acupuncture Stimulation of Paired Acupoints Can Relieve Sleep Disorder Possibly by Upregulating Pineal Melatonin Protein and Its Receptor mRNA Levels in the Suprachiasmatic Nucleus in Insomnia Rats

目的:观察针刺不同经脉穴位对失眠大鼠松果体褪黑素(MT)含量及视交叉上核(SCN)内褪黑素受体1(MT_1)、褪黑素受体2(MT_2)mRNA表达的影响,探讨固定局部穴位配伍不同经脉穴位之间作用的差异。方法:雄性SD大鼠随机分为空白组、模型组、百会+神门组、百会+三阴交组和百会+非经非穴组,每组12只。采用腹腔注射对氯苯丙氨酸混悬液建立失眠模型。治疗组每日针刺治疗1次,连续治疗7d。运用免疫组织化学法检测大鼠松果体MT的表达量,运用实时荧光定量PCR法检测大鼠SCN内MT_1、MT_2 mRNA的表达量。结果:与空白组比较,模型组大鼠松果体MT灰度值升高,SCN内MT_1、MT_2mRNA表达量降低(P<0.05,P<0.01)。与模型组比较,百会+神门组、百会+三阴交组松果体MT灰度值明显降低(P<0.05),百会+三阴交组、百会+神门组SCN内MT_1、MT_2mRNA表达量均明显升高(P<0.01)。各治疗组组间比较,百会+神门组MT_1mRNA表达量明显高于百会+非经非穴组(P<0.01),MT_2mRNA表达量明显高于百会+三阴交组及百会+非经非穴组(P<0.01)。结论:针刺治疗可以提高失眠大鼠松果体MT含量及SCN内MT_1、MT_2mRNA表达量,固定局部穴位配伍循经选穴可以提高腧穴配伍效应。

Objective To observe the effect of manual acupuncture stimulation of paired ＂Baihui＂ （GV 20） ＋＂Shenmen＂ （HT 7）, GV 20 ＋ ＂Sanyinjiao＂ （SP 6）, and GV 20 ＋ non-acupoint on expression of melatonine （MT） and suprachiasmatic melatonin receptor 1 （MT1） and melatonin receptor 2 （MT2） mRNAs in insomnia rats, so as to explore their action difference and the underlying mechanism in improving insomnia. Methods Male SD rats were randomly divided into normal control （n = 12）, model （n=8）, GV20＋HT7（n=12）, GV20＋SP6（n=11）, andGV20＋non-acupeint （n=10） groups. The insomnia model was established by intraperitoneal injection of Para-chlorophenylalanine suspension （50 mg/mL, 50 rag/100 g）, once daily for 2 days. The abovementioned acupoints GV 20, bilateral HT 7, SP 6 and non-acupoints （the midpoint between the elbow-tip and armpit on the medial side of the upper-arm） were punctured with filiferm needles and manipulated by rotating the needle for about 1 min which was repeated once again every 10 min during 30 minutes＇ needle-retaining. The treatment was conducted once daily for 7 days. The expression levels of MT immunoactivity in the conarium tissue, and MT1 and MT2 mRNAs of the suprachiasmatic nucleus （SCN） region were detected using immunohistochemistry and fluorescence quantitative real time-POR, respectively. Results After modeling, the expression levels of pineal MT （an increase of gray value means a decrease of immunoactivity）, SCN MT1 and MT2 mRNAs were notably down-regulated in the model group relevant to the normal control group （P〈0.05, P〈0.01 ）. Following the treatment, the down-regulated expression levels of MT protein, and MT1 and MT2 mRNAs were obviously reversed in the GV 20 ＋ HT 7, GV 20 ＋ SP 6 groups relevant to the model group （P〈0.05, P〈0.01）. The therapeutic effect of GV 20＋HT 7 was su-perior to that of GV 20 ＋ non-acupoint in up-regulating the expression of MT1 mRNA （P〈0.01）, and markedly superior to that of GV 20＋ SP 6 and GV 20＋ nen-acupoint in increasing the sleep duration and in up-regulating the expression of MT2 mRNA （P〈0.01 ）. No significant differences were found among the 3 treatment groups in up-regulating the expression of MT （ P〈0.05）, Conclusion Manual acupuncture stimulation of GV 20 ＋ HT 7 and GV 20 ＋ SP 6 can improve the sleep disorder in insomnia rats, which may be related to its effects in increasing the levels of pineal MT protein, and MT1 and MT2 mRNAs in hypothalamic SON.