针刺对抑郁症模型大鼠海马miR-152-3p、miR-145-5p及CaMKⅡ/NF-κB信号通路的影响OA

Objective:To observe the effects of the governor vessel-unblocking and mind-regulating acupuncture method on miR-152-3p,miR-145-5p,key molecules of the calcium/calmodulin-dependent protein kinase Ⅱ(CaMKⅡ)/nuclear factor(NF)-κB signaling pathway,and downstream inflammatory factors in cornu ammonis area 1(CA1)in chronic unpredictable mild stress(CUMS)model rats,thereby exploring the possible mechanism of the governor vessel-unblocking and mind-regulating acupuncture method in treating depression. Methods:Twelve of 48 Sprague-Dawley rats were randomly selected as a blank control group,and the other rats were subjected to CUMS for 28 consecutive days to prepare a depression rat model.The successful model rats were randomly divided into a model group,an acupuncture group,and a fluoxetine group,with 12 rats in each group.The blank control and model groups did not receive intervention.The acupuncture group received acupuncture at Baihui(GV20),Shuigou(GV26),Shenting(GV24),and Dazhui(GV14),6 d a week for a total of 4 weeks;the fluoxetine group was administered fluoxetine by gavage for 28 consecutive days.The behavioral changes of rats in each group after intervention were observed;hematoxylin-eosin staining and Nissl staining were used to observe the pathological morphology and neuronal damage of the hippocampal CA1 region.The levels of interleukin(IL)-1β,IL-6,tumor necrosis factor(TNF)-α,IL-10,and 5-hydroxytryptamine(5-HT)in the hippocampus and serum were detected by enzyme-linked immunosorbent assay.Relative expression levels of miR-152-3p,miR-145-5p,and mRNAs of CaMKⅡ and NF-κB p65 in the hippocampus were detected by reverse transcription real-time fluorescence quantitative polymerase chain reaction.The expression levels of CaMKⅡ,NF-κB p65,IL-1β,IL-6,TNF-α,and IL-10 in the hippocampal CA1 region were detected by immunofluorescence.Relative expression levels of CaMKⅡ and NF-κB p65 protein in the hippocampus were detected by Western blotting assay. Results:Compared with the blank control group,CUMS rats showed decreased numbers of squares crossed and upright standings and consumption rate of sucrose(P<0.01),and increased swimming immobility time(P<0.01);obvious inflammatory injury in hippocampal CA1 region;increased levels of IL-1β,IL-6,and TNF-α in the hippocampus and serum(P<0.01),and decreased levels of IL-10 and 5-HT(P<0.01);decreased relative expression levels of miR-152-3p and miR-145-5p in the hippocampus(P<0.01)and increased relative mRNA and protein expression levels of CaMKⅡ and NF-κB p65 in the hippocampus(P<0.01),increased mean fluorescence intensities of CaMKⅡ,NF-κB p65,IL-1β,IL-6,and TNF-α(P<0.01),and decreased mean fluorescence intensity of IL-10(P<0.01)in hippocampal CA1 area.Compared with the model group,rats in the acupuncture group and the fluoxetine group had increased numbers of squares crossed and upright standings and consumption rate of sucrose(P<0.01),and significantly reduced swimming immobility time(P<0.01);reduced neuroinflammatory reaction in hippocampal CA1 region;decreased levels of IL-1β,IL-6,and TNF-α(P<0.01),and increased levels of IL-10 and 5-HT(P<0.01)in the hippocampus and serum;increased relative expression levels of miR-152-3p and miR-145-5p in the hippocampus(P<0.01),and decreased relative mRNA and protein expression levels of CaMKⅡ and NF-κB p65 in the hippocampus(P<0.01);decreased mean fluorescence intensities of CaMKⅡ,NF-κB p65,IL-1β,IL-6,and TNF-α in hippocampal CA1 area(P<0.01),while increased mean fluorescence intensity of IL-10(P<0.01).Relative expression levels of miR-152-3p and miR-145-5p in rat hippocampus were negatively correlated with CaMKⅡ mRNA(-1<r<0,P<0.001),while CaMKⅡ mRNA was positively correlated with NF-κB p65 mRNA(0<r<1,P<0.001). Conclusion:The mechanism of the governor vessel-unblocking and mind-regulating acupuncture method in treating depression may be related to the upregulation of miR-152-3p and miR-145-5p expression levels in the hippocampus and the inhibition of neuritis responses caused by the CaMKⅡ/NF-κB signaling pathway in the CA1 region.