摩腹疗法对失眠大鼠食欲素及相关神经递质的影响

Effects of abdominal massage on orexin and related neurotransmitter in insomnia rats

目的观察摩腹疗法对失眠大鼠食欲素(Orexin)及相关神经递质的影响。方法将64只Wistar大鼠随机分为正常对照组、模型对照组、摩腹手法组、受体拮抗剂组,每组16只。模型对照组、摩腹手法组、受体拮抗剂组通过改良平台水环境法建立失眠大鼠模型,造模完成后摩腹手法组予摩腹疗法,受体拮抗剂组予食欲素受体拮抗剂10 mg/kg灌胃后再行摩腹疗法,正常对照组、模型对照组仅予外固定架束缚,均每日操作1次,连续2周。采用酶联免疫吸附法(ELISA)检测脑组织及血清食欲素A(Orexin-A)、5-羟色胺(5-HT)、多巴胺(DA)、去甲肾上腺素(NA)、γ-氨基丁酸(GABA)、乙酰胆碱(Ach)、组胺(HA)的水平。结果血清学指标:与正常对照组比较,模型对照组血清DA、NA、Ach、HA、Orexin-A含量均升高(P<0.05),5-HT、GABA含量均降低(P<0.05);与模型对照组比较,摩腹手法组和受体拮抗剂组DA、Ach含量均降低(P<0.05),5-HT含量均升高(P<0.05),受体拮抗剂组血清Orexin-A含量降低(P<0.05),摩腹手法组血清Orexin-A含量无明显变化(P>0.05)。摩腹手法组和受体拮抗剂组组间各检测指标比较差异无统计学意义(P>0.05)。脑组织指标:与正常对照组比较,模型对照组脑干DA、NA、Ach含量和下丘脑HA、Orexin-A含量均升高(P<0.05),脑干5-HT含量降低(P<0.05),摩腹手法组、受体拮抗剂组脑干Ach含量均升高(P<0.05),脑干5-HT含量均降低(P<0.05);与模型对照组比较,摩腹手法组脑干DA、NA、Ach含量和受体拮抗剂组脑干DA、NA含量和下丘脑Orexin-A含量均降低(P<0.05),脑干5-HT含量均升高(P<0.05)。摩腹手法组和受体拮抗剂组组间各检测指标比较差异无统计学意义(P>0.05)。结论摩腹疗法治疗失眠可能是通过降低外周血清和中枢神经系统的Orexin水平,进而兴奋5-HT能神经元,抑制NA、DA、Ach能神经元以促进睡眠和维持睡眠。

Objective To observe the effects of abdominal massage(AM)on orexin and related neurotransmitter in insomnia rats.Methods Totally 64 Wistar rats were randomly divided into the normal control group,the model control group,the AM group and the receptor blocker group,with 16 rats in each group.Apart from the normal control group,the model of insomnia rats was established in the other three groups through the modified platform method.After modeling,the rats in the AM group were given AM;the rats in the receptor blocker group were given 10 mg/kg orexin receptor antagonist by gavage,and then treatmeat with AM;the rats in the normal and model control groups were was only bound by external fixator;the treatment was performed once a day,continued for 2 weeks.The brain tissue,serum orexin-A,5-hydroxytryptamine(5-HT),dopamine(DA),norepinephrine(NA),gamma-aminobutyric acid(GABA),acetylcholine(Ach),histamine(HA)were detected using the enzyme-linked immunosorbent assay(ELISA).Results In the comparison of serological indicators among the groups,higher serum DA,NA,Ach,HA,orexin-A,and lower 5-HT,GABA were detected in the model control group than those in the normal control group(all P<0.05);decreased DA,Ach and increased 5-HT in the AM group and the receptor blocker group,and decreased orexin-A in the receptor blocker group were detected than those in the model control group(all P<0.05);the serum orexin-A in the AM group had no significant change compared with the model control group(P>0.05);the difference was not statistically significant in the each indicator between the AM group and the receptor blocker group(P>0.05).Compared with the normal control group,DA,NA,Ach in brain stem and HA,orexin-A in hypothalamus in the model control group were increased(P<0.05),5-HT in brain stem was decreased(P<0.05);Ach in brain stem in the AM group and the receptor blocker group was increased(P<0.05),5-HT in brain stem was decreased(P<0.05);compared with the model control group,DA,NA,Ach in brain stem and orexin-A in hypothalamus in the AM group and the receptor blocker group were decreased(P<0.05),while 5-HT in brain stem was increased(P<0.05);none of the each indicator differed significantly between the AM group and the receptor blocker group(P>0.05).Conclusion AM may stimulate 5-HT neurons and inhibit NA,DA and Ach neurons to promote sleep and maintain sleep by lowering the orexin level of peripheral blood and central nervous system.