针刺相对穴"内关与外关"对失眠大鼠海马及穴区肥大细胞脱颗粒及ICAM-1、SP表达的影响OA

Objective:To observe the effect of acupuncture intervention at complementary acupuncture points"Neiguan(PC6)and Waiguan(TE5)"on mast cell(MC)degranulation and intercellular adhesion molecule-1(ICAM-1)and substance P(SP)expression in the hippocampus and acupoint area of p-chlorophenylalanine(PCPA)-induced insomnia model Wistar rats.Methods:A total of 80 male Wistar rats were randomly selected,with 12 designated as the blank group.The remaining rats underwent intraperitoneal PCPA injection to establish an insomnia model,and 60 successfully modeled rats were re-randomized in-to a model group,single acupoint groups(single Neiguan group and single Waiguan group),and complementary acupuncture point groups(penetrating acupuncture group and opposing acupuncture group),with 12 rats in each group.Hippocampal morphological changes were observed via crystal violet staining.Hippocampal morphological changes were observed via crystal violet stai-ning.Acetylcholine iodide staining was used to visualize MC-vascular neural network structures in acupoint regions.Toluidine blue staining was used to observe MC counts and degranulation changes in the central hippocampus and peripheral acupoint tis-sues.Immunohistochemistry was used to detected ICAM-1,SP,and its receptor SPR(neurokinin 1 receptor,NK1R)protein ex-pression.RT-PCR and Western blot(WB)were used to detect ICAM-1 mRNA and protein expression in the central hippocampus and peripheral acupoint tissues.Results:Hippocampal neurons in the blank group exhibited regular arrangement,rounded mor-phology,and centrally located nuclei,whereas hippocampal neurons in the model group neurons showed irregular shapes,sparse cells,and nuclear displacement.The neuronal morphology in the complementary acupuncture point groups was closer to the blank group,with a more rounded shape and a higher proportion of centrally located nuclei.Compared to the blank group,model rats ex-hibited increased hippocampal MC counts and degranulation rates(P＜0.05).Compared to the model group,the complementary acupuncture point groups showed reduced hippocampal MC counts and MC degranulation rates,but elevated MC counts and MC degranulation rates in acupoint regions(P＜0.05).Immunohistochemistry results revealed that,compared to the blank group,the hippocampal and acupoint ICAM-1,SP,and SRP(NK1R)expression in the model group were significantly elevated.Compared to the model group,the complementary acupuncture point groups exhibited significantly reduced hippocampal protein expression of ICAM-1,SP,and SPR(NK1R)(P＜0.05),and significantly enhanced acupoint protein expression of these markers.RT-PCR and WB confirmed that,compared to the blank group,the hippocampal and acupoint ICAM-1 mRNA and protein in the model group increased;compared to the model group,the complementary acupuncture points showed reversed hippocampal ICAM-1 mRNA and protein expression(P＜0.05),and amplified acupoint ICAM-1 mRNA and protein expression(P＜0.05).Conclusion:Acupunc-ture at complementary acupuncture points"Neiguan and Waiguan"can improve sleep in the insomnia rat model,and its mecha-nism may be through the hippocampus and acupoint area tissues jointly regulating and promoting MC aggregation and degranula-tion to produce neurotransmitter changes,and in the acupoint area locally,SP and ICAM-1 and other substances contact and ad-here to the neurovascular system,forming a relatively stable MC vascular neural network structure.