單次阻力運動調控發炎機制探討：STAT3扮演之角色

Abstract

背景：長期的慢性發炎會增加罹患慢性疾病的風險。單次阻力運動可以增加抗發炎細胞激素，進而抑制發炎路徑達到抗發炎之效果。然而，現階段阻力運動相關研究仍多以觀察循環細胞激素濃度變化為主，對於阻力運動後對STAT3訊息傳遞路徑的反應尚待後續研究闡明。目的：一、探討單次阻力運動後健康年輕男性在循環免疫細胞數目及循環細胞激素濃度的影響。二、探討單次阻力運動後健康年輕男性在周邊血液單核細胞 (peripheral blood mononuclear cells, PBMCs) 中STAT3調控發炎相關路徑的影響。方法：本研究招募16名健康年輕男性作為研究對象，以隨機交叉設計，進行單次阻力運動 (resistance exercise, RE) 或安靜休息 (control, CON) 兩種實驗處理。於運動前、運動後立即、運動後2小時、運動後4小時及隔日 (24小時) 採集血液樣本。測量循環免疫細胞數量、IL-6、IL-10、TNF-α及PBMCs中的STAT3相關路徑。實驗數據以混合線性模型 (linear-mixed model) 進行統計考驗。結果：總白血球數量於RE後在運動後立即 (RE: 6.79 ± 2.07 ×106/mL; CON: 5.08 ± 1.06 ×106/mL)、運動後2小時 (RE: 7.51 ± 2.61 ×106/mL; CON: 5.59 ± 0.84 ×106/mL) 與運動後4小時 (RE: 7.43 ± 2.32 ×106/mL; CON: 5.86 ± 1.18 ×106/mL) 顯著高於CON (p< 0.05)。嗜中性球數目於RE在運動後2小時 (RE: 5.23 ± 2.77 ×106/mL; CON: 3.05 ± 1.06 ×106/mL) 與運動後4小時 (RE: 4.86 ± 2.5 ×106/mL; CON: 3.27 ± 1.42 ×106/mL) 顯著高於CON (p < 0.05)。淋巴球數目於RE在運動後立即 (RE: 1.89 ± 0.7 ×106/mL; CON: 1.24 ± 0.45 ×106/mL) 顯著高於CON (p < 0.05)。單核球數目於兩種處理間無顯著差異 (p > 0.05)。血漿IL-6濃度於RE在運動後立即 (RE: 1.25 ± 0.65 pg/mL; CON: 0.76 ± 0.41 pg/mL) 與運動後4小時 (RE: 4.31 ± 3.20 pg/mL; CON: 2.21 ± 0.67 pg/mL) 顯著高於CON (p < 0.05)。血漿IL-10濃度於RE在運動後立即 (RE: 1.02 ± 0.29 pg/mL; CON: 0.86 ± 0.26 pg/mL) 顯著高於CON (p < 0.05)。血漿TNF-ɑ濃度於兩種處理間無顯著差異 (p> 0.05)。PBMC細胞質中STAT3蛋白表現量於兩種處理間無顯著差異 (p > 0.05)。PBMCs中磷酸化STAT3α和STAT3β於RE在運動後4小時皆顯著高於CON (p < 0.05)。PBMCs中SOCS3蛋白表現量於RE在運動後立即、運動後2小時、運動後4小時及運動後24小時皆顯著高於CON (p < 0.05)。細胞核/細胞質STAT3蛋白表現量比值於兩種處理間無顯著差異 (p > 0.05)。PBMCs中細胞核SHIP1蛋白表現量於兩種處理間無顯著差異 (p > 0.05)。結論：單次阻力運動誘發IL-6和IL-10抗發炎細胞激素濃度上升以及活化PMBCs中STAT3訊息傳遞路徑。其中，IL-6濃度的上升與PBMCs中的STAT3磷酸化同時發生，可能說明阻力運動後對循環免疫細胞抗發炎效應的可能機制。

Background: Chronic inflammation increases the risk of developing and progressing several diseases. The anti-inflammatory effect of resistance exercise (RE) is mediated by increasing circulating anti-inflammatory cytokines to inhibit inflammatory pathways. However, previous studies mainly focused on changes in inflammation-related cytokines following acute RE. The activation of anti-inflammatory signaling pathways in immune cells after acute RE remains unclear. Purpose: 1) To examine the effect of acute RE on circulating white blood cell (WBC) counts and cytokines in healthy young male, 2) to investigate the impact on the anti-inflammatory signaling pathways within peripheral blood mononuclear cells (PBMCs) in healthy young males. Methods: Sixteen healthy young male completed two randomized crossover conditions (RE: resistance exercise; CON: sedentary control). Blood samples were collected at pre-exercise as well as immediately, 2 h, 4 h, and 24 h post-exercise. Circulating WBC counts, IL-6, IL-10, TNF-α, and STAT3 signaling pathway in PBMCs. Collected data were analyzed using linear-mixed model. Results: WBC counts increased in RE at immediately post-exercise (RE: 6.79 ± 2.07 ×106/mL; CON: 5.08 ± 1.06 ×106/mL), 2 h post-exercise (RE: 7.51 ± 2.61 ×106/mL; CON: 5.59 ± 0.84 ×106/mL), and 4 h post-exercise (RE: 7.43 ± 2.32 ×106/mL; CON: 5.86 ± 1.18 ×106/mL) compare with CON (p< 0.05). Higher neutrophil counts were observed in RE versus CON at 2 h post-exercise (RE: 5.23 ± 2.77 ×106/mL; CON: 3.05 ± 1.06 ×106/mL) and 4 h post-exercise (RE: 4.86 ± 2.5 ×106/mL; CON: 3.27 ± 1.42 ×106/mL; p < 0.05). Lymphocyte counts in RE were significant higher at immediately post-exercise compared with CON (RE: 1.89 ± 0.7 ×106/mL; CON: 1.24 ± 0.45 ×106/mL; p < 0.05). No differences in monocytes counts were observed between conditions (p < 0.05). Plasma IL-6 concentration increased in REat immediately post-exercise (1.25 ± 0.65 vs. 0.76 ± 0.41 pg/mL) and 4 h post-exercise (4.31 ± 3.20 vs. 2.21 ± 0.67 pg/mL) compared with CON (p< 0.05). Higher plasma IL-10 concentration were observed in RE versus CON at immediately post-exercise (1.02 ± 0.29 vs. 0.86 ± 0.26 pg/mL; p < 0.05). No differences in plasma TNF-ɑ concentration were observed between conditions (p < 0.05). Changes in total STAT3ɑ and STAT3β protein expression in cytoplasmic fractions of PBMCs were not different between conditions (p > 0.05). Phosphorylation of both STAT3ɑ and STAT3β in cytoplasmic fractions of PBMCs were increased in RE compared with CON at 4 hpost-exercise (p< 0.05). SOCS3 protein expression in cytoplasmic fractions of PBMCs was greater in RE than CON at immediately post-exercise, 2 h post-exercise, 4 h post-exercise, and 24 h post-exercise (p < 0.05). No differences in both STAT3 and SHIP1 protein expression in nuclear fractions of PBMCs were observed following RE (p > 0.05). Conclusion: Acute RE promotes an increase in circulating WBC and neutrophil counts. In addition, acute RE elevates anti-inflammatory cytokines IL-6 and IL-10 and activating the STAT3 signaling pathway in PBMCs. The increase in IL-6 coincided with the activation of STAT3 signaling in PBMCs, highlighting a potential mechanism by which RE may exert anti-inflammatory actions in circulating immune cells.