膜微粒抑制缺氧无血清诱导的骨髓间充质干细胞的凋亡
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摘要
目的:明确心肌梗死后血液来源的膜微粒(MPs)抑制缺氧无血清诱导的骨髓间充质干细胞 (BMSCs)凋亡的作用并探讨其相关机制。方法:利用结扎SD大鼠左冠脉前降支的方法来制作急性心肌梗死模型,然后提取血液中的MPs。提取培养SD大鼠的BMSCs,并建立缺氧无血清的干细胞凋亡模型。分别按照不同浓度(即0.5μg/mL、1 μg/mL和2 μg/mL)的MPs对干细胞进行预处理,分别利用Hoechst染色、Tunel检测、AnnexinV/PI流式细胞学、Western blot检测caspase-3来揭示MPs抑制BMSCs的凋亡作用,并用Akt信号通路的通路抑制剂AZD5363来初步探讨其抗凋亡的相关机制。结果:大鼠心肌梗死后血液来源的MPs能够有效抑制缺氧无血清诱导的BMSCs的凋亡(P<0.01);并且其抗凋亡作用呈现出浓度依赖性,0.5 μg/mL浓度的MPs亦能达到显著抗凋亡作用(P<0.05)。其机制主要是通过激活Akt信号通路来实现。结论:心肌梗死后血液来源的MPs能有效抑制缺氧无血清诱导的BMSCs的凋亡。
Objective: To investigate the effects of myocardial infarction plasma-derived microparticles(MPs) on the hypoxia and serum deprivation induced apoptosis of bone marrow mesenchymal stem cells(BMSCs). Methods: Myocardial infarction(MI) was created by left anterior descending artery ligation in Lewis rats. The plasma-derived MPs were isolated 24 hours later from the blood. In order to study the protective effects of MPs on BMSCs under the conditions of hypoxia and serum deprivation(hypoxia/SD). BMSCs were pretreated with different concentrations(0.5 μg/mL, 1 μg/mL, and 2 μg/mL) of MPs before the apoptosis was induced. Cell apoptosis was evaluated by using flow cytometry of Annexin V/PI staining, Hoechst staining and the Tunel assay. Expression of cleavage of caspase-3 were assessed by Western blotting. The Akt pathway inhibitor AZD5363 was used to reveal the mechanism of MPs inhibiting BMSCs apoptosis. Results: The MPs derived from the myocardial infarction rats could remarkably prevent BMSCs from Hypoxia/SD induced apoptosis through activating Akt signaling pathway, and its anti-apoptosis effect showed a concentration dependence(0.5-2 μg/mL)(P<0.01). Conclusion: MPs derived from myocardial infarction could effectively inhibit hypoxia/SD induced apoptosis of BMSCs.
Objective: To investigate the effects of myocardial infarction plasma-derived microparticles(MPs) on the hypoxia and serum deprivation induced apoptosis of bone marrow mesenchymal stem cells(BMSCs). Methods: Myocardial infarction(MI) was created by left anterior descending artery ligation in Lewis rats. The plasma-derived MPs were isolated 24 hours later from the blood. In order to study the protective effects of MPs on BMSCs under the conditions of hypoxia and serum deprivation(hypoxia/SD). BMSCs were pretreated with different concentrations(0.5 μg/mL, 1 μg/mL, and 2 μg/mL) of MPs before the apoptosis was induced. Cell apoptosis was evaluated by using flow cytometry of Annexin V/PI staining, Hoechst staining and the Tunel assay. Expression of cleavage of caspase-3 were assessed by Western blotting. The Akt pathway inhibitor AZD5363 was used to reveal the mechanism of MPs inhibiting BMSCs apoptosis. Results: The MPs derived from the myocardial infarction rats could remarkably prevent BMSCs from Hypoxia/SD induced apoptosis through activating Akt signaling pathway, and its anti-apoptosis effect showed a concentration dependence(0.5-2 μg/mL)(P<0.01). Conclusion: MPs derived from myocardial infarction could effectively inhibit hypoxia/SD induced apoptosis of BMSCs.
引文
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[8]陈棉,王珏,耿志敏,等.骨髓间充质干细胞条件膜微粒对心脏成纤维细胞的影响[J].温州医科大学学报, 2016,46(9):625-628.
[9]BERNAL-MIZRACHI L, JY W, JIMENEZ J J, et al. High levelsofcirculatingendothelialmicroparticlesinpatients with acute coronary syndromes[J]. Am Heart J, 2003, 145(6):962-970.
[10]GHOLAMIN S, PASDAR A, KHORRAMI M S, et al. The potential for circulating microRNAs in the diagnosis of myocardial infarction:a novel approach to disease diagnosis and treatment[J]. Curr Pharm Des, 2016, 22(3):397-403.
[11]QUESENBERRY P J, ALIOTTA J M. Cellular phenotype switching and microvesicles[J]. Adv Drug Deliv Rev, 2010,62(12):1141-1148.
[12]QUESENBERRY P J, DOONER M S, ALIOTTA J M. Stem cellplasticityrevisited:thecontinuummarrowmodeland phenotypic changes mediated by microvesicles[J]. Exp Hematol, 2010, 38(7):581-592.
[2]RODRIGO S F, MANN I, RAMSHORST J VAN, et al.Safetyandefficacyofpercutaneousintramyocardialbone marrow cell injection for chronic myocardial ischemia:longterm results[J]. J Interv Cardiol, 2017, 30(5):440-447.
[3]CHEN Y, ZHAO Y, CHEN W, et al. MicroRNA-133 overexpression promotes the therapeutic efficacy of mesenchymal stem cells on acute myocardial infarction[J]. Stem Cell Res Ther, 2017, 8(1):268.
[4]RAPOSO G, STOORVOGEL W. Extracellular vesicles:exosomes, microvesicles, and friends[J]. J Cell Biol, 2013,200(4):373-383.
[5]BENAMEUR T, ANDRIANTSITOHAINA R, MARTINEZ MC. Therapeuticpotentialofplasmamembrane-derived microparticles[J]. Pharmacol Rep, 2009, 61(1):49-57.
[6]金培峰,张浩,陆地,等.雪旺细胞移植改善心肌梗死后副交感/交感神经重塑[J].温州医科大学学报, 2016, 46(1):13-18.
[7]SIONIS A, SUADES R, SANS-ROSELLO J, et al. Circulating microparticles are associated with clinical severity of persistent ST-segment elevation myocardial infarction complicated with cardiogenic shock[J]. Int J Cardiol, 2018, 258:249-256.
[8]陈棉,王珏,耿志敏,等.骨髓间充质干细胞条件膜微粒对心脏成纤维细胞的影响[J].温州医科大学学报, 2016,46(9):625-628.
[9]BERNAL-MIZRACHI L, JY W, JIMENEZ J J, et al. High levelsofcirculatingendothelialmicroparticlesinpatients with acute coronary syndromes[J]. Am Heart J, 2003, 145(6):962-970.
[10]GHOLAMIN S, PASDAR A, KHORRAMI M S, et al. The potential for circulating microRNAs in the diagnosis of myocardial infarction:a novel approach to disease diagnosis and treatment[J]. Curr Pharm Des, 2016, 22(3):397-403.
[11]QUESENBERRY P J, ALIOTTA J M. Cellular phenotype switching and microvesicles[J]. Adv Drug Deliv Rev, 2010,62(12):1141-1148.
[12]QUESENBERRY P J, DOONER M S, ALIOTTA J M. Stem cellplasticityrevisited:thecontinuummarrowmodeland phenotypic changes mediated by microvesicles[J]. Exp Hematol, 2010, 38(7):581-592.