PDF(Pubmed)
Abstract：
UNASSIGNED: To study the mechanism by which conditioned medium of bone marrow mesenchymal stem cells (BMSCs-CM) facilitates the transition of pro-inflammatory polarized microglia to an anti-inflammatory phenotype.
UNASSIGNED: BV2 cells, a mouse microglia cell line, were transformed into a pro-inflammatory phenotype using lipopolysaccharide. The expression of phenotypic genes in BV2 cells was detected using real-time quantitative PCR (RT-qPCR). Enzyme-linked immunosorbent assay was used to measure inflammatory cytokine levels in BV2 cells co-cultured with BMSCs-CM. The expressions of mitophagy-associated proteins were determined using western blot. The mitochondrial membrane potential and ATP levels in BV2 cells were measured using JC-1 staining and an ATP assay kit, respectively. Additionally, we examined the proliferation, apoptosis, and migration of C8-D1A cells, a mouse astrocyte cell line, co-cultured with BV2 cells.
UNASSIGNED: After co- culture with BMSCs -CM, the mRNA expression of tumor necrosis factor-α (TNF-α) and inducible nitric oxide synthase significantly decreased in pro-inflammatory BV2 cells, whereas the expression of CD206 and arginase-1 significantly increased. Moreover, TNF-α and interleukin-6 levels significantly decreased, whereas transforming growth factor-β and interleukin-10 levels significantly increased. Furthermore, co-culture with BMSCs-CM increased mitophagy-associated protein expression, ATP levels, mitochondrial and lysosomal co-localization in these cells and decreased reactive oxygen species levels. Importantly, BMSCs-CM reversed the decrease in the proliferation and migration of C8-D1A cells co-cultured with pro-inflammatory BV2 cells and inhibited the apoptosis of C8-D1A cells.
UNASSIGNED: BMSCs-CM may promote the transition of polarized microglia from a pro-inflammatory to an anti-inflammatory phenotype by regulating mitophagy and influences the functional state of astrocytes.
UNASSIGNED: BV2 cells, a mouse microglia cell line, were transformed into a pro-inflammatory phenotype using lipopolysaccharide. The expression of phenotypic genes in BV2 cells was detected using real-time quantitative PCR (RT-qPCR). Enzyme-linked immunosorbent assay was used to measure inflammatory cytokine levels in BV2 cells co-cultured with BMSCs-CM. The expressions of mitophagy-associated proteins were determined using western blot. The mitochondrial membrane potential and ATP levels in BV2 cells were measured using JC-1 staining and an ATP assay kit, respectively. Additionally, we examined the proliferation, apoptosis, and migration of C8-D1A cells, a mouse astrocyte cell line, co-cultured with BV2 cells.
UNASSIGNED: After co- culture with BMSCs -CM, the mRNA expression of tumor necrosis factor-α (TNF-α) and inducible nitric oxide synthase significantly decreased in pro-inflammatory BV2 cells, whereas the expression of CD206 and arginase-1 significantly increased. Moreover, TNF-α and interleukin-6 levels significantly decreased, whereas transforming growth factor-β and interleukin-10 levels significantly increased. Furthermore, co-culture with BMSCs-CM increased mitophagy-associated protein expression, ATP levels, mitochondrial and lysosomal co-localization in these cells and decreased reactive oxygen species levels. Importantly, BMSCs-CM reversed the decrease in the proliferation and migration of C8-D1A cells co-cultured with pro-inflammatory BV2 cells and inhibited the apoptosis of C8-D1A cells.
UNASSIGNED: BMSCs-CM may promote the transition of polarized microglia from a pro-inflammatory to an anti-inflammatory phenotype by regulating mitophagy and influences the functional state of astrocytes.
摘要：
研究骨髓间充质干细胞(BMSCs-CM)条件培养基促进促炎极化小胶质细胞向抗炎表型转变的机制。
■BV2细胞，小鼠小胶质细胞系，使用脂多糖转化为促炎表型。采用实时定量PCR(RT-qPCR)检测BV2细胞中表型基因的表达。酶联免疫吸附试验用于测量与BMSCs-CM共培养的BV2细胞中的炎性细胞因子水平。免疫印迹法检测线粒体自噬相关蛋白的表达。使用JC-1染色和ATP测定试剂盒测量BV2细胞中的线粒体膜电位和ATP水平，分别。此外,我们检查了扩散，凋亡，和C8-D1A细胞的迁移，小鼠星形胶质细胞系，与BV2细胞共培养。
■与BMSCs-CM共培养后，促炎症BV2细胞中肿瘤坏死因子-α(TNF-α)和诱导型一氧化氮合酶的mRNA表达显著降低,而CD206和精氨酸酶-1的表达明显增加。此外，TNF-α和白细胞介素-6水平显著降低,而转化生长因子-β和白细胞介素-10水平显著升高。此外，与BMSCs-CM共培养增加线粒体自噬相关蛋白的表达，ATP水平,这些细胞中的线粒体和溶酶体共定位，并降低了活性氧水平。重要的是,BMSCs-CM逆转了与促炎BV2细胞共培养的C8-D1A细胞增殖和迁移的减少，并抑制了C8-D1A细胞的凋亡。
■BMSCs-CM可能通过调节线粒体自噬促进极化小胶质细胞从促炎表型向抗炎表型转变，并影响星形胶质细胞的功能状态。
■BV2细胞，小鼠小胶质细胞系，使用脂多糖转化为促炎表型。采用实时定量PCR(RT-qPCR)检测BV2细胞中表型基因的表达。酶联免疫吸附试验用于测量与BMSCs-CM共培养的BV2细胞中的炎性细胞因子水平。免疫印迹法检测线粒体自噬相关蛋白的表达。使用JC-1染色和ATP测定试剂盒测量BV2细胞中的线粒体膜电位和ATP水平，分别。此外,我们检查了扩散，凋亡，和C8-D1A细胞的迁移，小鼠星形胶质细胞系，与BV2细胞共培养。
■与BMSCs-CM共培养后，促炎症BV2细胞中肿瘤坏死因子-α(TNF-α)和诱导型一氧化氮合酶的mRNA表达显著降低,而CD206和精氨酸酶-1的表达明显增加。此外，TNF-α和白细胞介素-6水平显著降低,而转化生长因子-β和白细胞介素-10水平显著升高。此外，与BMSCs-CM共培养增加线粒体自噬相关蛋白的表达，ATP水平,这些细胞中的线粒体和溶酶体共定位，并降低了活性氧水平。重要的是,BMSCs-CM逆转了与促炎BV2细胞共培养的C8-D1A细胞增殖和迁移的减少，并抑制了C8-D1A细胞的凋亡。
■BMSCs-CM可能通过调节线粒体自噬促进极化小胶质细胞从促炎表型向抗炎表型转变，并影响星形胶质细胞的功能状态。
