"引阳入阴"法针刺对失眠模型大鼠海马Nek7/NLRP3/Caspase-1/IL-1β信号通路的影响OA

Objective To observe the effect of acupuncture with the method of"inducing Yang into Yin"on the signal pathway of NIMA-related protein kinase 7/oligonucleotide-binding domain-like receptor protein 3/cysteine aspartate proteolytic enzyme-1/interleukin-1β(Nek7/NLRP3/Caspase-1/IL-1β)in hippocampus of insomnia model rats,and to explore the mechanism of acupuncture in improving insomnia.Methods Forty SPF healthy male SD rats were randomly divided into blank group(10 rats)and model group(30 rats).The insomnia model was established by intraperitoneal injection of p-chlorophenylalanine(PCPA).Randomly select 30 rats that successfully established the model and divide them into model group,acupuncture group and Western medicine group,with 10 rats in each group.The acupuncture group was given acupuncture by the method of"inducing Yang into Yin",and the points were"Baihui(GV20)""Waiguan(TE5)(double)""Neiguan(PC6)(double)""Weishu(BL21)(double)""Zhongwan(CV12)"and"Taixi(KI3)(double)"once a day for 7 days.The Western medicine group was given estazolam solution once a day for 7 days.The insomnia behavior of rats in each group was evaluated by pentobarbital sodium correction test and open field test.HE matoxylin-eosin staining was used to observe the pathological changes of hippocampus in rats.The contents of IL-1β and IL-18 in hippocampus of rats were detected by ELISA.The mRNA expressions of Nek7,NLRP3,apoptosis-related spot-like protein(ASC),Caspase-1,IL-1β and IL-18 were detected by RT-qPCR.The expressions of Nek7,NLRP3,ASC,Caspase-1,IL-1β and IL-18 in hippocampus were detected by Western blot.Results Compared with the blank group,the weight of rats in each group decreased significantly(P<0.01).The duration of sleep latency was significantly prolonged and the sleep time was significantly shortened(P<0.01).In the open field experiment,the total distance and time of exercise increased obviously(P<0.01),and the residence time in the central area decreased(P<0.05).The hippocampus tissue atrophied,the number of neurons decreased obviously with disordered arrangement,some nuclei contracted and deformed,cytoplasm concentrated and stained deeply,and punctate necrotic foci and micro-bleeding appeared in some areas.The contents of IL-1β and IL-18 in hippocampus of rats increased significantly(P<0.01).The mRNA and protein expression levels of Nek7,NLRP3,ASC,Caspase-1,IL-1β and IL-18 all increased significantly(P<0.01).Compared with the model group,the body weight of the Western medicine group and the acupuncture group increased significantly(P<0.05,P<0.01).The sleep latency was shortened(P<0.01)and the sleep time was obviously prolonged(P<0.01).The total distance and time of exercise decreased significantly(P<0.01),and the stay time in the central area increased significantly(P<0.05,P<0.01).The number of neurons is close to the normal level,the cell arrangement is basically orderly,the nucleoli is clearly visible,and only punctate necrotic foci and individual cytoplasm concentration are occasionally seen.The contents of IL-1β and IL-18 in hippocampus decreased significantly(P<0.01).The mRNA and protein expression levels of Nek7,NLRP3,ASC,Caspase-1,IL-1β,IL-18 decreased in different degrees(P<0.05,P<0.01).Compared with Western medicine group,the total distance and time of exercise in acupuncture group were significantly reduced(P<0.05).The expression of ASC,Caspase-1,IL-1β and IL-18 mRNA decreased(P<0.05,P<0.01).The expression levels of Nek7,NLRP3,ASC,Caspase-1,IL-1β and IL-18 decreased(P<0.05,P<0.01).Conclusion Acupuncture with the method of"inducing Yang into Yin"can improve the sleep quality of insomnia rats and reduce the inflammatory reaction of hippocampus,and its mechanism may be related to the regulation of Nek7/NLRP3/caspase-1/IL-1β signal pathway.