4.2 Article

Mechanical Stretch Promotes Macrophage Polarization and Inflammation via the RhoA-ROCK/NF-κB Pathway

期刊

BIOMED RESEARCH INTERNATIONAL
卷 2022, 期 -, 页码 -

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HINDAWI LTD
DOI: 10.1155/2022/6871269

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资金

  1. Natural Science Foundation of China [81673995]
  2. Priority Academic Program Development of Jiangsu Higher Education Institutions (Integration of Chinese and Western Medicine)
  3. Qing Lan Project of Jiangsu Province [[2018] 12]
  4. Natural Science Foundation of Jiangsu Province [BK20211300]
  5. Postgraduate Research & Practice Innovation Program of Jiangsu Province [KYCX17_1308, KYCX21_1683]

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Macrophages play a crucial role in inflammatory diseases, and mechanical load can influence their function and enhance inflammatory responses. This study found that mechanical stretch can regulate macrophage polarization and enhance inflammation in vitro.
Macrophages play an essential role in the pathogenesis of most inflammatory diseases. Recent studies have shown that mechanical load can influence macrophage function, leading to excessive and uncontrolled inflammation and even systemic damage, including cardiovascular disease and knee osteoarthritis. However, the molecular mechanism remains unclear. In this study, murine RAW264.7 cells were treated with mechanical stretch (MS) using the Flexcell-5000T Tension System. The expression of inflammatory factors and cytokine release were measured by RT-qPCR, ELISA, and Western blotting. The protein expression of NF-kappa B p65, I kappa b-alpha, p-I kappa b-alpha, RhoA, ROCK1, and ROCK2 was also detected by Western blotting. Then, Flow cytometry was used to detect the proportion of macrophage subsets. Meanwhile, Y-27632 dihydrochloride, a ROCK inhibitor, was added to knockdown ROCK signal transduction in cells. Our results demonstrated that MS upregulated mRNA expression and increased the secretion levels of proinflammatory factors iNOS, IL-1 beta, TNF-alpha, and IL-6. Additionally, MS significantly increased the proportion of CD11b+CD86+ and CD11b+CD206+ subsets in RAW264.7 macrophages. Furthermore, the protein expression of RhoA, ROCK1, ROCK2, NF-kappa B p65, and I kappa B-alpha increased in MS-treated RAW264.7 cells, as well as the IL-6 and iNOS. In contrast, ROCK inhibitor significantly blocked the activation of RhoA-ROCK and NF-kappa B pathway, decreased the protein expression of IL-6 and iNOS, reduced the proportion of CD11b+CD86+ cells subpopulation, and increased the proportion of CD11b+CD206+ cell subpopulation after MS. These data indicate that mechanical stretch can regulate the RAW264.7 macrophage polarization and enhance inflammatory responses in vitro, which may contribute to activation the RhoA-ROCK/NF-kappa B pathway.

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