Abstract：
Peripheral blood mononuclear cells (PBMC), sourced autologously, offer numerous advantages when procured: easier acquisition process, no in vitro amplification needed, decreased intervention and overall increased acceptability make PBMC an attractive candidate for cell therapy treatment. However, the exact mechanism by which PBMC treat diseases remains poorly understood. Immune imbalance is the pathological basis of many diseases, with macrophages playing a crucial role in this process. However, research on the role and mechanisms of PBMC in regulating macrophages remains scarce. This study employed an in vitro co-culture model of PBMC and RAW264.7 macrophages to explore the role and mechanisms of PBMC in regulating macrophages. The results showed that the co-culturing led to decreased expression of inflammatory cytokines and increased expression of anti-inflammatory cytokines in RAW264.7 or in the culture supernatant. Additionally, the pro-inflammatory, tissue matrix-degrading M1 macrophages decreased, while the anti-inflammatory, matrix-synthesizing, regenerative M2 macrophages increased in both RAW264.7 and monocytes within PBMC. Moreover, co-cultured macrophages exhibited a significantly decreased p-STAT1/STAT1 ratio, while the p-STAT6/STAT6 ratio significantly increased. This suggests that PBMC may inhibit M1 macrophage polarization by blocking STAT1 signaling cascades and may promote M2 macrophage polarization through the activation of STAT6 signaling cascades. Overall, this study sheds light on the role and mechanism of PBMC in regulating macrophages. Moreover, it was found that monocytes within co-cultured PBMC differentiated into M2 macrophages in the presence of macrophages. This finding provides experimental evidence for the use of PBMC in treating inflammatory diseases, especially macrophage-depleting inflammatory diseases such as osteoarthritis.
摘要：
外周血单核细胞（PBMC），自主采购，在采购时提供了许多优势：更容易的采购过程，不需要体外扩增，干预措施的减少和总体可接受性的增加使PBMC成为细胞疗法治疗的有吸引力的候选者.然而,PBMC治疗疾病的确切机制仍然知之甚少。免疫失衡是许多疾病的病理基础,巨噬细胞在这个过程中起着至关重要的作用。然而,关于PBMC在调节巨噬细胞中的作用和机制的研究仍然很少。本研究采用PBMC和RAW264.7巨噬细胞的体外共培养模型来探讨PBMC在调节巨噬细胞中的作用和机制。结果表明,共培养导致RAW264.7或培养上清液中炎性细胞因子的表达降低和抗炎细胞因子的表达增加。此外,促炎，组织基质降解M1巨噬细胞减少，而抗炎，矩阵合成，再生M2巨噬细胞在RAW264.7和PBMC内的单核细胞中均增加。此外，共培养的巨噬细胞表现出显著降低的p-STAT1/STAT1比值，而p-STAT6/STAT6比值显著增加。这表明PBMC可以通过阻断STAT1信号传导级联来抑制M1巨噬细胞极化,并且可以通过激活STAT6信号传导级联来促进M2巨噬细胞极化。总的来说,本研究揭示了PBMC在调节巨噬细胞中的作用和机制。此外，发现在巨噬细胞存在下,共培养的PBMC内的单核细胞分化成M2巨噬细胞。这一发现为使用PBMC治疗炎症性疾病提供了实验证据。尤其是消耗巨噬细胞的炎性疾病，如骨关节炎。
