Aerobic Training Inhibits Hippocampal Inflammation by Activating the Hippocampus TLR4/miR-223/NLRP3 Signaling Pathway Axis in Mice with CUMS-induced Depression
(2.Yichun University, Yichun, China 336000)
【Abstract】 Objective: The purpose of this study was to explore the effects of aerobic training on the hippocampal inflammation by activating the hippocampal TLR4/miR-223/NLRP3 signal pathway in mice with chronic unpredictable mild stress (CUMS)-induced depression. Methods: Sixty 8-week-old male C57BL/6 mice were divided into control group (CG), CUMS model group (MG), CUMS model plus exercise group (ME), TLR4 inhibitor group (TG), and TLR4 inhibitor plus exercise group (TE) (n = 12). Except CG, the CUMS was molded for mice of the other four groups with chronic stress for 28 days. The mice in both ME group and TE group were trained using a treadmill at a moderate intensity for 8 weeks. The mice in TG group and TE group were received intraperitoneal injection of TAK-242, an inhibitor of TLR4, for 4 weeks. The neurobehavioral scores were tested after the 8-week training for all the mice. The samples were collected the day after their last training. The levels of blood IL-1β and IL-10 were measured using ELISA kit; the morphology of Nissl body in hippocampus was detected by Nissl staining; the positive expression areas of TLR4, IL-1β, IL-10, and NF-κB proteins were detected by immunohistochemistry; the mRNA expression levels of NLRP3, TLR4, IL-1β, IL-10, NF-κB, and miR-223 in hippocampus were examined by RT-PCR; the protein expression levels of TLR4, IL-1β, and IL-10 in hippocampus were tested by Western blot. Results: The mice with CUMS showed significantly decreased neurobehavioral function, significantly increased inflammatory cytokines such as IL-1β in blood, and the appearance of Nissl body pyknosis. The 8-week aerobic training significantly improved the hippocampus function in the mice with CUMS, reduced the level of IL-1β in blood, Nissl body pyknosis, and the desperate behaviors, and increased the mouse desire to survive. High-throughput sequencing and correlation analysis of miRNA and mRNA showed that there was a targeting regulatory relationship between miR-223 and NLRP3, which was influenced by exercise intervention. The expressions of inflammatory cytokines such as NLRP3, TLR4, IL-1β, and NF-κB were down-regulated while that of the anti-inflammatory cytokine IL-10 was up-regulated in ME mice. The expressions of NF-κB and NLRP3 were down-regulated and that of miR-223 was up-regulated, and the anti-inflammatory effect was increased in TG mice. The consistent anti-inflammatory effect was also found in TE mice. Conclusion: Aerobic training significantly reduced the expression of TLR4 in the hippocampus of mice with CUMS-induced depression while activated the TLR4/miR-223/NLRP3 pathway axis, thus improving the hippocampus function to inhibit inflammation and enhance the repair of hippocampal inflammatory injury.
【Keywords】 depression; chronic unpredictable mild stress; microRNA-223; aerobic training; hippocampus; TLR4/miR-223/NLRP3;
(Translated by PXJ1)
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