1.Effect of Abdominal Massage Combined with Thumb-tack Needling for Subcutaeous Embedding on Ventrolateral Preoptic Nucleus of Hypothalamic Sleep Homeostasis System in Rats with Anxiety Insomnia
Si-Gan QIU ; Xue-Feng ZHANG ; Gulaisaer·AIKEBAIER ; Xu-Sheng LU ; Lei GUO ; Xing-Ping ZHANG ; Jun-Chang LIU
Journal of Guangzhou University of Traditional Chinese Medicine 2024;41(6):1549-1559
Objective To observe the effect of abdominal massage combined with thumb-tack needling for subcutaeous embedding on sleep homeostasis system in rats with anxiety insomnia.Methods Forty rats were randomly divided into normal group,model group,abdominal massage group,thumb-tack needling for subcutaeous embedding group and abdominal massage plus thumb-tack needling for subcutaeous embedding group,with 8 rats in each group.Except for the normal group,the rats in the other groups were used to replicate the model of anxiety insomnia by multi-factor compound stimulation.After the corresponding intervention,Morris water maze test was used to detect the level of learning and memory.Open field test was used to detect the degree of anxiety stress.Hematoxylin-eosin(HE)staining was used to observe the pathological changes of hypothalamic ventral lateral preoptic nucleus(VLPO)neurons.Immunohistochemistry,real-time quantitative polymerase chain reaction(qRT-PCR)and Western Blot were used to detect the protein and mRNA expression of N-methyl-D-aspartate(NMDA)receptor subunits NR1,NR2B and calmodulin kinase Ⅱ(CaMK Ⅱ)in hypothalamic VLPO area,respectively.Results Compared with the normal group,the daytime anxiety symptoms of the rats in the model group were aggravated,the sleep latency was prolonged and the duration was shortened(P<0.01).The average total swimming distance and average escape latency of the water maze directional navigation experiment were increased(P<0.01).The number of crossing the hidden platform and the retention time of the target quadrant in the space exploration experiment were decreased(P<0.01).The movement distance,the number of central grid crossings and the retention time of the central grid in the open field experiment were significantly reduced(P<0.01).There was no significant difference in the modification frequency and the number of uprights(P>0.05).Neurons in the VLPO brain region showed pathological damage.The protein and mRNA expression levels of NR1 and CaMK Ⅱ were decreased(P<0.01)in VLPO brain region,and the protein and mRNA expression levels of NR2B were increased(P<0.01).Compared with the model group,the level of learning and memory in the water maze test and the degree of anxiety stress in the open field test were significantly restored in the abdominal massage group,the thumb-tack needling for subcutaeous embedding group and the abdominal massage combined with thumb-tack needling for subcutaeous embedding group(P<0.05 or P<0.01),the neuronal damage in the VLPO brain region was improved,the protein and mRNA expression levels of NR1,CaMK Ⅱ were increased(P<0.05 or P<0.01),and the protein and mRNA expression levels of NR2B were decreased(P<0.05 or P<0.01).The improvement effect of the above indexes in the abdominal massage plus thumb-tack needling for subcutaeous embedding group was superior to that in the abdominal massage group or thumb-tack needling for subcutaeous embedding group(P<0.05 or P<0.01).Conclusion Abdominal massage combined with thumb-tack needling for subcutaeous embedding can promote sleep and anti-anxiety in rats with anxiety insomnia.The related mechanism may be related to adjusting the dynamic balance between NR1/NR2B in VLPO brain area and up-regulating the expression level of CaMK Ⅱ,improving the function of neurons in VLPO brain area,and then restoring the regulation of sleep homeostasis system.