Human Mesenchymal Stem Cell Hydrogen Sulfide Production Critically Impacts the Release of Other Paracrine Mediators After Injury.
Human Mesenchymal Stem Cell Hydrogen Sulfide Production Critically Impacts the Release of Other Paracrine Mediators After Injury.
复制标题
人类间充质干细胞硫化氢的产生严重影响受伤后其他旁分泌介质的释放。
DOI：
10.1016/j.jss.2020.04.014
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发表时间：
2020
期刊：
影响因子：
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通讯作者：
Olson,KennethR
中科院分区：
文献类型：
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作者：
Markel,TroyA;Drucker,NatalieA;Jensen,AmandaR;Olson,KennethR
背景间充质干细胞（mesenchymal stem cells，MSCs）用于缺血性疾病的治疗是一种新的方法。硫化氢（H2S）是一种重要的旁分泌介质，其从MSC释放以促进血管生成和血管舒张。三种酶，胱硫醚-β-合成酶（CBS）、胱硫醚-γ-裂解酶（CSE）和3-巯基丙酮酸-硫转移酶（MPST），主要负责H2S的产生。然而，目前还不清楚这些酶如何影响其他关键生长因子和趋化因子的产生。我们假设，负责H2S的生产在人类骨髓间充质干细胞的酶也将严重调节其他生长因子和chemokines.Materials和methodsHuman骨髓间充质干细胞转染CBS，MPST，CSE，或阴性对照小干扰RNA。通过聚合酶链反应确认酶的敲低。将细胞以每孔100，000个细胞接种在12孔板中，并用肿瘤坏死因子-α（TNF-α; 50 ng/mL）、脂多糖（LPS; 200 ng/mL）或5%缺氧刺激24 h。收集上清液，并通过多重珠粒测定法测量细胞因子。结果TNF-α、LPS、缺氧均能有效刺激MSCs增殖。在TNF-α刺激过程中，当CSE被敲低时，粒细胞集落刺激因子（GCSF）、表皮生长因子、成纤维细胞生长因子、粒细胞/单核细胞集落刺激因子（GMCSF）、血管内皮生长因子和干扰素γ诱导蛋白10均显著升高（P< 0.05）。在LPS刺激下，MPST的敲除更容易增加GCSF和表皮生长因子，而减少GMCSF（P< 0.05）。CBS基因敲低可降低缺氧后GCSF、成纤维细胞生长因子、GMCSF和血管内皮生长因子的产生（P< 0.05）。结论应激刺激下，MSCs中产生H2S的酶也参与了其他干细胞旁分泌介质的产生。因此，重编程MSC以内源性产生更多的H2S作为治疗干预也可能严重影响其他旁分泌介质，这可能会改变所需的有益效果。
BackgroundThe use of mesenchymal stem cells (MSCs) for treatment during ischemia is novel. Hydrogen sulfide (H2S) is an important paracrine mediator that is released from MSCs to facilitate angiogenesis and vasodilation. Three enzymes, cystathionine-beta-synthase (CBS), cystathionine-gamma-lyase (CSE), and 3-mercaptopyruvate-sulfurtransferase (MPST), are mainly responsible for H2S production. However, it is unclear how these enzymes impact the production of other critical growth factors and chemokines. We hypothesized that the enzymes responsible for H2S production in human MSCs would also critically regulate other growth factors and chemokines.Materials and methodsHuman MSCs were transfected with CBS, MPST, CSE, or negative control small interfering RNA. Knockdown of enzymes was confirmed by polymerase chain reaction. Cells were plated in 12-well plates at 100,000 cells per well and stimulated with tumor necrosis factor-α (TNF-α; 50 ng/mL), lipopolysaccharide (LPS; 200 ng/mL), or 5% hypoxia for 24 h. Supernatants were collected, and cytokines measured by multiplex beaded assay. Data were compared with the Mann–WhitneyU-test, andP< 0.05 was significant.ResultsTNF-α, LPS, and hypoxia effectively stimulated MSCs. Granulocyte colony-stimulating factor (GCSF), epidermal growth factor, fibroblast growth factor, granulocyte/monocyte colony-stimulating factor (GMCSF), vascular endothelial growth factor, and interferon gamma-inducible protein 10 were all significantly elevated when CSE was knocked down during TNF-α stimulation (P< 0.05). Knockdown of MPST during LPS stimulation more readily increased GCSF and epidermal growth factor but decreased GMCSF (P< 0.05). CBS knockdown decreased production of GCSF, fibroblast growth factor, GMCSF, and vascular endothelial growth factor (P< 0.05) after hypoxia.ConclusionsThe enzymes that produce H2S in MSCs are also responsible for the production of other stem cell paracrine mediators under stressful stimuli. Therefore, reprogramming MSCs to endogenously produce more H2S as a therapeutic intervention could also critically impact other paracrine mediators, which may alter the desired beneficial effects.