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Effect of cannabinoid receptor 2 on autism-like behaviors inmice by regulating microglial function
- TIAN Wenru, YANG Bilin, HU Jingyi, LIU Shidan, ZOU Mingying, WU Lijie, SUN Caihong
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2025, 33(1):
62-70.
DOI: 10.11852/zgetbjzz2024-1205
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Objective To explore the regulatory effect of cannabinoid receptor 2 (CB2R) on microglial function and its impact on autism-like behaviors in mice, in order to provide scientific basis for the etiology of ASD. Methods In vitro experiment, BV2 microglia were divided into control group, LPS group, and AM1241 group. RT-PCR was used to detect mRNA expression levels of inflammatory factors. Western Blot was used to measure the proteins expression of pyroptosis marker. In vivo experiments, the 8-week-old mice were divided into WT group and CB2R knockout mice (CB2R-/- group). Immunofluorescence staining was used to evaluate the activation of microglia. RT-PCR and western blot were used to detect mRNA and proteins expression levels of pyroptosis marker. Anxiety, restricted and repetitive behaviors, and social function were evaluated by open field, self-grooming, burying marble, and three-chamber test. Results In vitro experiments, compared with the control group, the mRNA levels of pro-inflammatory factors TNF-α, IL-6, and CCL2 in the LPS group were increased, while anti-inflammatory factors Arg-1 and IL-13 were decreased (P<0.05). Compared with the LPS group, the mRNA levels of TNF-α, IL-6, and CCL2 in the AM1241 group were decreased, while Arg-1, IL-10, and IL-13 were increased (P<0.000 1). Compared with the control group, the protein expression of NLRP3, GSDMD, GSDMD-N, caspase-1, and cleaved caspase-1 were significantly increased in the LPS group. After intervention with AM1241, the above protein levels were significantly reduced (P<0.05). In vivo experiments, compared with the WT group, the CD86+/Iba1+ ratios in the hippocampal CA1 and CA3 regions were increased, while the CD206+/Iba1+ ratios were decreased in the CB2R-/- group (P<0.01). The mRNA expression of NLRP3, GSDMD, caspase-1, IL-1β, and IL-18 in CB2R-/- group was significantly increased (t=2.20, 3.12, 2.55, 2.54, 2.30, P<0.05). The protein expression levels of NLRP3, GSDMD, GSDMD-N, caspase-1, and cleaved caspase-1 in CB2R-/- group were significantly increased (t=7.43, 2.87, 3.86, 3.99, 6.55, P<0.05). The CB2R-/- group showed a significant increase in the numbers of moved distance, self-grooming time and marbles buried in mice, a significant decrease in the resting time and social preference index (P<0.05). Conclusion CB2R is involved in regulating the activation and pyroptosis of microglia, affecting neuroinflammatory responses, thereby leading to autism-like behaviors in mice, which suggests that CB2R might be a potential therapeutic target for ASD.