间充质干细胞上清液对巨噬细胞生物学行为的影响

    Effect of mesenchymal stem cells supernatant liquid on the biological behavior of macrophages

    • 摘要:
      目的: 探讨间充质干细胞(MSCs)培养的上清液(MSCs–SL)对巨噬细胞脂质蓄积、极化、炎症及凋亡等生物学行为的影响。
      方法: MSCs–SL与THP–1巨噬细胞共培养,采用MTT法检测巨噬细胞活性的影响,油红O染色观察细胞中脂质蓄积情况,实时荧光定量PCR技术检测肿瘤坏死因子–α(TNF–α)、白细胞介素(IL)–1β、主动脉分化簇206(CD206)和人精氨酸酶1(ARG–1)的mRNA表达量,缺口末端标记(TUNEL)染色观察巨噬细胞凋亡变化。
      结果: 20% MSCs–SL可以提高LPS诱导的THP–1巨噬细胞活性(F = 63.70,P < 0.01)。20% MSCs–SL处理细胞后,未见对巨噬细胞脂质蓄积显著影响(F = 2.26,P > 0.05)。MSCs–SL共培养组显著上调炎症状态下TNF–α(F = 125.90,P < 0.01)、IL–1β(F = 165.00,P < 0.01)的mRNA水平和M1极性状态下TNF–α(t = 12.60,P < 0.01)、IL–1β(t = 8.77,P < 0.01)的mRNA水平,并下调M2极性状态下的CD206的mRNA水平(t = 6.19,P < 0.05)。MSCs–SL处理可显著抑制巨噬细胞凋亡。
      结论: MSCs–SL对巨噬细胞脂质蓄积无明显影响,但促进巨噬细胞炎症和M1极化,抑制巨噬细胞M2极化及凋亡。

       

      Abstract:
      Objective To investigate the effects of supernatant liquid from cultured mesenchymal stem cells (MSCs) on the lipid accumulation, polarisation, inflammation and apoptosis of macrophages.
      Methods The mesenchymal stem cells supernatant liquid (MSCs-SL) was co-cultured with THP-1 macrophages. The THP-1 macrophages activity were detected by MTT assay. The lipid accumulation of THP-1 macrophages were detected by oil red O staining. Real-time PCR (qRT-PCR) was used to detect the expression levels of TNF-α, IL-1β, CD206 and ARG-1. The apoptosis of THP-1 macrophages were detected by Tunel staining.
      Results The 20% MSCs-SL could enhanced the lipopolysaccharide-induced macrophage activity (F = 63.70, P < 0.01). After treating the cells with 20% MSCs-SL, no significant effect on lipid accumulation in macrophages was observed (F = 2.26, P > 0.05). The MSCs-SL co-culture group significantly upregulated the mRNA levels of TNF-α (F = 125.90, P < 0.01) and IL-1β (F = 165.00, P < 0.01) under inflammatory conditions and TNF-α (t = 12.60) and IL-1β (t = 8.77, P < 0.01) under M1 polarity conditions, and down-regulated the mRNA level of CD206 in the M2 polar state (t = 6.19, P < 0.05). MSCs-SL treatment could significantly inhibit the macrophage apoptosis.
      Conclusions MSCs-SL has no significant effect on lipid accumulation in macrophages, but it can promote the macrophage inflammation and M1 polarization, and inhibit the M2 polarization and apoptosis of macrophages.

       

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