多发性硬化症(MS)以中枢神经系统的炎症细胞激活和浸润引起的脱髓鞘化和神经元丧失为特征。巨噬细胞极化在经典多发性硬化症实验性自身免疫脑脊髓炎(EAE)的发病机制中发挥重要作用。本研究调查了Fasudil对巨噬细胞的影响，并检测了Fasudil修饰巨噬细胞在实验性自身免疫脑脊髓炎中治疗潜力。我们发现Fasudil诱导了巨噬细胞由促炎性M1型转化为抗炎性M2型，表现为诱导型一氧化氮合酶/一氧化氮、白细胞介素-12和CD16/32的表达减少及精氨酸酶-1、白细胞介素-10、CD14和CD206的表达增加。这与Rho激酶活性的抑制、Toll样受体、核因子κB和丝裂原活化蛋白激酶信号通路组分的表达减少，以及产生促炎性细胞因子肿瘤坏死因子-α、白细胞介素-1β和白细胞介素-6有关。重要的是，Fasudil修饰巨噬细胞有效减少了实验性自身免疫脑脊髓炎的影响，导致疾病发病较晚、症状评分较低、体重减轻较少和脱髓鞘减少，与未修饰巨噬细胞相比。此外，Fasudil修饰巨噬细胞降低了CD4+ T细胞和F4/80+巨噬细胞上的CD16/32、诱导型一氧化氮合酶和白细胞介素-12的表达，同时增加了CD4+ T细胞和F4/80+巨噬细胞上白细胞介素-10的表达、精氨酸酶-1、CD206和白细胞介素-10的表达，从而改善了免疫调节并降低了炎症。这些发现表明，Fasudil修饰巨噬细胞可能通过诱导M2巨噬细胞极化和抑制炎症反应来治疗实验性自身免疫脑脊髓炎，从而为多发性硬化症的细胞免疫治疗提供了新的见解。

Multiple sclerosis is characterized by demyelination and neuronal loss caused by inflammatory cell activation and infiltration into the central nervous system. Macrophage polarization plays an important role in the pathogenesis of experimental autoimmune encephalomyelitis, a traditional experimental model of multiple sclerosis. This study investigated the effect of Fasudil on macrophages and examined the therapeutic potential of Fasudil-modified macrophages in experimental autoimmune encephalomyelitis. We found that Fasudil induced the conversion of macrophages from the pro-inflammatory M1 type to the anti-inflammatory M2 type, as shown by reduced expression of inducible nitric oxide synthase/nitric oxide, interleukin-12, and CD16/32 and increased expression of arginase-1, interleukin-10, CD14, and CD206, which was linked to inhibition of Rho kinase activity, decreased expression of toll-like receptors, nuclear factor-κB, and components of the mitogen-activated protein kinase signaling pathway, and generation of the pro-inflammatory cytokines tumor necrosis factor-α, interleukin-1β, and interleukin-6. Crucially, Fasudil-modified macrophages effectively decreased the impact of experimental autoimmune encephalomyelitis, resulting in later onset of disease, lower symptom scores, less weight loss, and reduced demyelination compared with unmodified macrophages. In addition, Fasudil-modified macrophages decreased interleukin-17 expression on CD4+ T cells and CD16/32, inducible nitric oxide synthase, and interleukin-12 expression on F4/80+ macrophages, as well as increasing interleukin-10 expression on CD4+ T cells and arginase-1, CD206, and interleukin-10 expression on F4/80+ macrophages, which improved immune regulation and reduced inflammation. These findings suggest that Fasudil-modified macrophages may help treat experimental autoimmune encephalomyelitis by inducing M2 macrophage polarization and inhibiting the inflammatory response, thereby providing new insight into cell immunotherapy for multiple sclerosis.