髓源性抑制细胞对脂多糖诱导的血管内皮细胞凋亡的保护作用
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摘要
目的探讨髓源性抑制细胞(MDSCs)对LPS诱导的内皮细胞凋亡是否有保护作用。方法采用流式分选技术从脓毒症患者外周血中分离MDSCs,将其与LPS预处理过的人脐带静脉内皮细胞(HUVECs)共培养设为实验组,HUVECs未经任何处理设为空白组,HUVECs经LPS刺激设为对照组。通过Hoechst染色观察各组细胞形态学上的变化,通过CCK-8以及流式测凋亡检测各组内皮细胞增殖、凋亡的情况。结果选择用浓度为10μg/mL的LPS刺激HUVEC 6 h构建脓毒症细胞模型。实验组内皮细胞增殖活性优于对照组(OD值:1.018 vs. 0.852,P=0.008),低于空白组(OD值:1.018 vs. 1.214,P=0.002);实验组凋亡率低于对照组(凋亡率:12.90%vs. 18.65%,P <0.001),高于空白组(凋亡率:12.90%vs.18.65%,P <0.001)。结论 MDSCs能促进LPS诱导的内皮细胞增殖、减少其凋亡,对内皮细胞的损伤及凋亡具有保护作用。
Objective To study whether the protective effect of myeloid-derived suppressor cells(MDSCs on LPS-induced apoptosis of endothelial cells. Methods Human umbilical vein endothelial cells(HUVECs)pretreated by LPS and then co-cultured with MDSCs separated from peripheral blood of patients with sepsis in flow sorting technique was set as experimental group;HUVECs without any treatment was set as blank group;HUVECs were stimulated by LPS was set as control group. The morphological changes of cells in each group were observed by Hoechst staining. The effects of proliferation and apoptosis of endothelial cells were detected by CCK-8 and flow cytometry. Results The cellular model of sepsis was constructed LPS stimulating HUVECs for 6 h with LPS concentration of 10μg/mL. Endothelial cell proliferation activity in the experimental group was better than in the control group(OD value:1.018 vs. 0.852,P = 0.008),lower than that of the blank group(OD value:1.018 vs.1.214,P = 0.002);The apoptosis rate in experimental group was lower than that in control group(apoptosis rate:12.90% vs. 18.65%,P < 0.001),higher than the blank group(apoptosis rate:12.90% vs. 18.65%,P < 0.001).Conclusion MDSCs can promote the proliferation of LPS-induced endothelial cells and reduce their apoptosis,which has protective effect on endothelial cells injury and apoptosis.
Objective To study whether the protective effect of myeloid-derived suppressor cells(MDSCs on LPS-induced apoptosis of endothelial cells. Methods Human umbilical vein endothelial cells(HUVECs)pretreated by LPS and then co-cultured with MDSCs separated from peripheral blood of patients with sepsis in flow sorting technique was set as experimental group;HUVECs without any treatment was set as blank group;HUVECs were stimulated by LPS was set as control group. The morphological changes of cells in each group were observed by Hoechst staining. The effects of proliferation and apoptosis of endothelial cells were detected by CCK-8 and flow cytometry. Results The cellular model of sepsis was constructed LPS stimulating HUVECs for 6 h with LPS concentration of 10μg/mL. Endothelial cell proliferation activity in the experimental group was better than in the control group(OD value:1.018 vs. 0.852,P = 0.008),lower than that of the blank group(OD value:1.018 vs.1.214,P = 0.002);The apoptosis rate in experimental group was lower than that in control group(apoptosis rate:12.90% vs. 18.65%,P < 0.001),higher than the blank group(apoptosis rate:12.90% vs. 18.65%,P < 0.001).Conclusion MDSCs can promote the proliferation of LPS-induced endothelial cells and reduce their apoptosis,which has protective effect on endothelial cells injury and apoptosis.
引文
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[15]CRIPPS J G,GORHAM J D.MDSC in autoimmunity[J].Int Immunopharmacol,2011,11(7):789-793.
[16]CHEN S,AKBAR S M,ABE M,et al.Immunosuppressive functions of hepatic myeloid-derived suppressor cells of normal mice and in a murine model of chronic hepatitis B virus[J].Clin Exp Immunol,2011,166(1):134-142.
[17]PEREIRA W F,RIBEIRO-GOMES F L,GUILLERMO L V,et al.Myeloid-derived suppressor cells help protective immunity to Leishmania major infection despite suppressed T cell responses[J].J Leukoc Biol,2011,90(6):1191-1197.
[18]KU A W,MUHITCH J B,POWERS C A,et al.Tumor-induced MDSC act via remote control to inhibit L-selectin-dependent adaptive immunity in lymph nodes[J].ELife,2016,5.pii:e17375.
[19]DANELLI L,FROSSI B,PUCILLO C E.Mast cell/MDSC a liaison immunosuppressive for tumor microenvironment[J].Oncoimmunology,2015,4(4):e1001232.
[20]BRUDECKI L,FERGUSON D A,MCCALL C E,et al.Myeloidderived suppressor cells evolve during sepsis and can enhance or attenuate the systemic inflammatory response[J].Infect Immun,2012,80(6):2026-2034.
[21]MERX M W,LIEHN E A,JANSSENS U,et al.HMG-CoAreductase inhibitor simvastatin profoundly improves survival in a murine model of sepsis[J].Circulation,2004,109(21):2560-2565.
[22]符晖,罗琼,谭思,等.辛伐他汀对脓毒症内皮细胞的保护作用[J].中华急诊医学杂志,2013(9):994-999.
[23]ZOSO A,MAZZA E M,BICCIATO S,et al.Human fibrocytic myeloid-derived suppressor cells express IDO and promote tolerance via Treg-cell expansion[J].Eur J Immunol,2014,44(11):3307-3319.
[24]李艳丽.外周T细胞淋巴瘤患者外周血中MDSCs比例及其临床意义的研究[D].桂林医学院,2015.
[2]POSCHKE I,KIESSLING R.On the armament and appearances of human myeloid-derived suppressor cells[J].Clin Immunol,2012,144(3):250-268.
[3]SANDER L E,SACKETT S D,DIERSSEN U,et al.Hepatic acute-phase proteins control innate immune responses during infection by promoting myeloid-derived suppressor cell function[J].Journal Exp Med,2010,207(7):1453-1464.
[4]DERIVE M,BOUAZZA Y,ALAUZET C,et al.Myeloidderived suppressor cells control microbial sepsis[J].Intensive Care Med,2012,38(6):1040-1049.
[5]DELANO M J,THAYER T,GABRILOVICH S,et al.Sepsis induces early alterations in innate immunity that impact mortality to secondary infection[J].J Immunol,2011,186(1):195-202.
[6]NOEL G,WANG Q,OSTERBURG A.et al.A ribonucleotide reductase inhibitor reverses burn-induced inflammatory defects[J].Shock,2010,34(5):535-544
[7]胡晓光,童荔,刘勇军,等.髓源性抑制细胞对脓毒症小鼠的保护作用[J].实用医学杂志,2014,30(11):1690-1692.
[8]SINGER M,DEUTSCHMAN C S,SEYMOUR C W,et al.The third international consensus definitions for sepsis and septic shock(Sepsis-3)[J].JAMA,2016,315(8):801-810.
[9]MABUCHI S,MATSUMOTO Y,KAWANO M,et al.Uterine cervical cancer displaying tumor-related leukocytosis:A distinct clinical entity with radioresistant feature[J].J Natl Cancer Inst,2014,106(7):766-776.
[10]林宁,邓家骏,付朋,等.人脑胶质瘤中Fas系统与其患者外周血中MDSC表达的相关性[J].中国临床神经外科杂志,2011,16(2):91-93.
[11]CUENCA A G,DELANO M J,KELLY-SCUMPIA K M,et al.A paradoxical role for myeloid-derived suppressor cells in sepsis and trauma[J].Mol Med,2011,17(3-4):281-292.
[12]CUENCA A G,MOLDAWER L L.Myeloid-derived suppressor cells in sepsis:Friend or foe?[J].Intensive Care Med,2012,38(6):928-930.
[13]MUNERA V,POPOVIC P J,BRYK J,et al.Stat 6-dependent induction of myeloid derived suppressor cells after physical injury regulates nitric oxide response to endotoxin[J].Ann Surg,2010,251(1):120-126.
[14]MAKARENKOVA V P,BANSAL V,MATTA B M,et al.CD11b+/Gr-1+myeloid suppressor cells cause T cell dysfunction after traumatic stress[J].J Immunol,2006,176(4):2085-2094.
[15]CRIPPS J G,GORHAM J D.MDSC in autoimmunity[J].Int Immunopharmacol,2011,11(7):789-793.
[16]CHEN S,AKBAR S M,ABE M,et al.Immunosuppressive functions of hepatic myeloid-derived suppressor cells of normal mice and in a murine model of chronic hepatitis B virus[J].Clin Exp Immunol,2011,166(1):134-142.
[17]PEREIRA W F,RIBEIRO-GOMES F L,GUILLERMO L V,et al.Myeloid-derived suppressor cells help protective immunity to Leishmania major infection despite suppressed T cell responses[J].J Leukoc Biol,2011,90(6):1191-1197.
[18]KU A W,MUHITCH J B,POWERS C A,et al.Tumor-induced MDSC act via remote control to inhibit L-selectin-dependent adaptive immunity in lymph nodes[J].ELife,2016,5.pii:e17375.
[19]DANELLI L,FROSSI B,PUCILLO C E.Mast cell/MDSC a liaison immunosuppressive for tumor microenvironment[J].Oncoimmunology,2015,4(4):e1001232.
[20]BRUDECKI L,FERGUSON D A,MCCALL C E,et al.Myeloidderived suppressor cells evolve during sepsis and can enhance or attenuate the systemic inflammatory response[J].Infect Immun,2012,80(6):2026-2034.
[21]MERX M W,LIEHN E A,JANSSENS U,et al.HMG-CoAreductase inhibitor simvastatin profoundly improves survival in a murine model of sepsis[J].Circulation,2004,109(21):2560-2565.
[22]符晖,罗琼,谭思,等.辛伐他汀对脓毒症内皮细胞的保护作用[J].中华急诊医学杂志,2013(9):994-999.
[23]ZOSO A,MAZZA E M,BICCIATO S,et al.Human fibrocytic myeloid-derived suppressor cells express IDO and promote tolerance via Treg-cell expansion[J].Eur J Immunol,2014,44(11):3307-3319.
[24]李艳丽.外周T细胞淋巴瘤患者外周血中MDSCs比例及其临床意义的研究[D].桂林医学院,2015.