不同极化的巨噬细胞通过调节 PACAP 的表达来影响 NSPCs 的活力和生长。
Differently polarized macrophages affect the viability and growth of NSPCs by regulating the expression of PACAP.
机构信息
Department of Immunology, School of Medicine, Wuhan University, Wuhan 430071, China.
Department of Orthopaedics, Renmin Hospital, Wuhan University, Wuhan City 430060, China.
出版信息
Neuropeptides. 2017 Oct;65:114-119. doi: 10.1016/j.npep.2017.07.003. Epub 2017 Jul 12.
UNLABELLED
Objective To explore the influence of differently polarized macrophages, M1 or M2, to viability and growth of NSPCs and its possible mechanism, especially the role of pituitary adenylate cyclase-activating polypeptide (PACAP) in it.
METHOD
Spinal cord-derived NSPCs were co-cultured with M1 or M2 through a transwell system. Growth of NSPCs in both groups was observed through an inverted microscope within 3days. NSPCs viability of each group, represented as intracellular ATP levels, was measured using the Cellular ATP Kit HTS following co-culture for 24h. PACAP levels in the co-cultured NSPCs was alleviated with immunofluorescence and Western blot analysis.
RESULTS
Morphologically NSPCs demonstrated a long spindle shape with short sprout on 3rd day when cultured together with M2. NSPCs cultured with M1 showed a small circle or oval shape with no obvious sprout. Expression of PACAP was observed in NSPCs co-cultured with M2 through immunofluorescence. In contrast, NSPCs did not demonstrate PACAP staining in the presence of M1 or cultured alone. PACAP in the NSPCs co-cultured with M2 was upregulated significantly compared with that co-cultured with M1 according to Western blot method. The relative ATP level of NSPCs co-cultured with M1 was markedly decreased while that with M2 was elevated significantly. That trend could be relieved by exogenous PACAP or PACAP 6-38. Viability change of NSPCs by M1/M2 correlated with apoptosis.
CONCLUSION
Differently polarized macrophages could affect the growth and viability of NSPCs by regulating the expression of PACAP.
未加标签
目的 探索不同极化的巨噬细胞（M1 或 M2）对神经干细胞（NSPCs）活力和生长的影响及其可能的机制，尤其是垂体腺苷酸环化酶激活肽（PACAP）在其中的作用。
方法
通过 Transwell 系统将脊髓源性 NSPCs 与 M1 或 M2 共培养。在共培养 3 天内，通过倒置显微镜观察两组 NSPCs 的生长情况。共培养 24 小时后，使用细胞 ATP 检测试剂盒 HTS 测量每组 NSPCs 的活力，以细胞内 ATP 水平表示。通过免疫荧光和 Western blot 分析减轻共培养 NSPCs 中的 PACAP 水平。
结果
当与 M2 共培养时，第 3 天 NSPCs 形态呈长梭形，短突起。与 M1 共培养的 NSPCs 呈小圆或椭圆形，无明显突起。通过免疫荧光观察到与 M2 共培养的 NSPCs 中存在 PACAP 表达。相比之下，在存在 M1 或单独培养的情况下，NSPCs 中未观察到 PACAP 染色。根据 Western blot 方法，与 M1 共培养的 NSPCs 中 PACAP 的表达明显上调，而与 M2 共培养的 NSPCs 中 PACAP 的表达明显上调。与 M1 共培养的 NSPCs 的相对 ATP 水平明显降低，而与 M2 共培养的 NSPCs 的相对 ATP 水平显著升高。该趋势可通过外源性 PACAP 或 PACAP 6-38 缓解。M1/M2 对 NSPCs 活力的影响与细胞凋亡相关。
结论
不同极化的巨噬细胞可通过调节 PACAP 的表达来影响 NSPCs 的生长和活力。
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