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 (CaMK II)/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 CaMK II and NF-κB p65 in the hippocampus were detected by reverse transcription real-time fluorescence quantitative polymerase chain reaction. The expression levels of CaMK II, 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 CaMK II 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 CaMK II and NF-κB p65 in the hippocampus (P<0.01), increased mean fluorescence intensities of CaMK II, 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 CaMK II and NF-κB p65 in the hippocampus (P<0.01); decreased mean fluorescence intensities of CaMK II, 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 CaMK II mRNA (-1<r<0, P<0.001), while CaMK II 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 CaMK II/NF-κB signaling pathway in the CA1 region.

Effects of acupuncture on miR-152-3p, miR-145-5p, and CaMK II/NF-κB signaling pathway in hippocampus of depression model rats

期刊论文
发表日期：

2026年6月23日

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Yanbiao Zhao,
Xiaoge Song,
Peiyang Sun,
Ling Zou,
Tingting Qian,
Yifan Chu,
Tao Jiang

摘要

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 (CaMK II)/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 CaMK II and NF-κB p65 in the hippocampus were detected by reverse transcription real-time fluorescence quantitative polymerase chain reaction. The expression levels of CaMK II, 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 CaMK II 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 CaMK II and NF-κB p65 in the hippocampus (P<0.01), increased mean fluorescence intensities of CaMK II, 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 CaMK II and NF-κB p65 in the hippocampus (P<0.01); decreased mean fluorescence intensities of CaMK II, 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 CaMK II mRNA (-1<r<0, P<0.001), while CaMK II 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 CaMK II/NF-κB signaling pathway in the CA1 region.