从5-HT1A、5-HT2A后信号通路探讨针刺治疗PCPA失眠机制

Mechanism study on the acupuncture treatment of PCPA insomnia through the post-receptor signal pathway of 5-HT1A and 5-HT2A

为探明针刺5-HT1A、5-HT2A后信号通路的影响,以及其与改善PCPA失眠中枢5-HT障碍机制的关系,将48只PCPA失眠大鼠随机均分为3组,并与16只正常鼠对照。正常组、模型组无特殊处理,针刺组予针刺脐内环合失眠穴方干预,非穴组针刺非穴处理,治疗6天后,处死动物、分离海马并匀浆,拟ELISA检测5-HT、PLCβ和cAMP含量,WB检测Gαi/o、Gαq/11表达,统计学分析5-HT与cAMP、PLCβ相关性。结果PCPA失眠大鼠海马5-HT、cAMP明显降低,PLCβ明显升高,针刺组5-HT、cAMP含量显著升高,PLCβ含量显著降低;各组内5-HT与cAMP呈正相关性、与PLCβ呈负相关性;模型组Gαq/11显著下降,针刺显示显著上调作用,各组间Gαi/o表达比较无统计意义;实验提示该针法能良性调控5-HT1A/Gαi/o/cAMP、5-HT2A/Gαq/11/PLCβ信号通路起改善PCPA失眠作用,该作用与改善中枢5-HT障碍机制有一定的相关性。

In order to observe the effect of acupuncture on the post-receptor signal pathway of 5-HT1A and 5-HT2A,and to explore the relationship between acupuncture effect on PCPA insomnia and the 5-HT antagonizing mechanism,48 insomnia rats induced by PCPA were randomly divided into 3 groups and were compared with the control group of 16 rats.The control group and the model group received no special treatment,the acupuncture group received the intervention of needling in the encircled acupoints of umbilicus and the insomnia points,while the non-acupoint group veceived the acupuncture at where there were no acupoints.Stripping and homogenating the hippocampus after 6 days of treatment,we detected the concentration of 5-HT,PLCβand cAMP with Elisa,explored the expression of Gαi/o and Gαq/11 with western blot,and used statistical methods to analysis the possible correlation between 5-HT,cAMP and PLCβ.Experimental results showed that the concentration of 5-HT and cAMP in the model group were significantly decreased and the concentration of PLCβwere significantly increased in the hippocampus of insomnia rats made by PCPA,while in acupuncture group,the concentration of 5-HT,cAMP were up-regulated and the PLCβwas down-regulated by acupuncture.In each group,the 5-HT and cAMP were positive correlated and the 5-HT and PLCβwere negatively correlated.The Gαq/11 was significantly decreased in the model group,but increased in acupuncture group.And there was no statistical significance of the Gαi/o among all groups.Experimental results indicate that needling in the encircled acupoints of umbilicus and the insomnia points could ameliorate 5-HT1A/Gαi/o/cAMP and 5-HT2A/Gαq/11/PLCβsignal pathway to improve the insomnia induced by PCPA,which is related to the mechanism of improving central 5-HT disorder.