镁对子痫前期患者内皮祖细胞增殖、黏附和凋亡的影响
|
摘要
背景:镁离子可通过自由基清除、细胞膜稳定、炎症抑制等作用降低血管内皮细胞对不利刺激的敏感性,调节血管内皮功能,但镁离子对内皮祖细胞的作用尚不清楚。目的:探讨镁对子痫前期患者内皮祖细胞增殖、黏附和凋亡的影响。方法:纳入2016年3月至2018年10月河北省人民医院产科17例子痫前期产妇与15名正常妊娠产妇,从两组产妇脐血中分离培养内皮祖细胞。将每例子痫前期产妇脐血来源内皮祖细胞分4组干预:子痫前期组以PBS干预,低、中、高浓度镁离子组分别以含4,8,16 mmol/L镁离子的培养基干预;正常妊娠产妇脐血来源内皮祖细胞以PBS干预,作为对照组。干预24 h后,检测各组细胞的增殖、黏附、凋亡、炎症因子(白细胞介素6、白细胞介素8、肿瘤坏死因子α与单核细胞趋化蛋白1)及氧化应激(超氧化物歧化酶、丙二醛)水平。实验方案已获河北省人民医院伦理委员会批准,伦理审批号:2013临审13号。结果与结论:①与对照组比较,子痫前期组细胞增殖与黏附数量明显降低(P <0.05),凋亡率升高(P <0.05),Bax和Active caspase-3蛋白表达升高(P <0.05),Bcl-2蛋白表达降低(P <0.05),白细胞介素6、白细胞介素8、肿瘤坏死因子α与单核细胞趋化蛋白1、丙二醛水平升高(P <0.05),超氧化物歧化酶活性降低(P <0.05);②与子痫前期组比较,镁离子干预各组细胞增殖与黏附数量升高(P <0.05),凋亡率降低(P <0.05),Bax和Active caspase-3蛋白表达降低(P <0.05),Bcl-2蛋白表达升高(P <0.05),炎症因子水平明显下降(P <0.05),超氧化物歧化酶活性升高(P <0.05),丙二醛水平降低(P <0.05),并且高浓度镁离子组改善程度优于低、中浓度镁离子组(P <0.05);③结果表明,镁离子可促进子痫前期患者体外内皮祖细胞的增殖和黏附,抑制炎性因子的分泌,降低凋亡率和氧化应激水平。
BACKGROUND: Magnesium ions can reduce the sensitivity of vascular endothelial cells to adverse stimuli and regulate vascular endothelial function by scavenging free radicals, stabilizing cell membranes and inhibiting inflammation. However, the effect of magnesium ions on endothelial progenitor cells is still unclear.OBJECTIVE: To explore the effects of magnesium on the proliferation, adhesion and apoptosis of endothelial progenitor cells in patients with preeclampsia.METHODS: From March 2016 to October 2018, 17 preeclampsia pregnant patients and 15 normal pregnant women who were admitted in the Obstetrics Department of Hebei General Hospital were enrolled. Endothelial progenitor cells were isolated and cultured from umbilical cord blood in the two groups of pregnant women. Endothelial progenitor cells isolated from each preeclampsia pregnant patient were divided into four groups: preeclampsia group was intervened with PBS, and low, middle and high concentration magnesium ion groups were intervened with the medium containing 4, 8, and 16 mmol/L magnesium ions, respectively. Endothelial progenitor cells from normal pregnant women were treated with PBS as control group. After 24 hours of intervention, cell proliferation, adhesion, apoptosis, inflammation(interleukin-6,interleukin-8, tumor necrosis factor-α, monocyte chemoattractant protein-1 levels) and oxidative stress(superoxide dismutase and malondialdehyde levels) were detected. The study protocol was approved by the Ethics Committee of Hebei General Hospital with the approval No. 2013 linshen13.RESULTS AND CONCLUSION:(1) Compared with the control group, the cell proliferation and adhesion of the preeclampsia group were significantly decreased(P < 0.05); the apoptotic rate significantly increased(P < 0.05); the expression of Bax and Active Caspase-3 increased,and the expression of Bcl-2 decreased(both P < 0.05); the levels of interleukin-6, interleukin-8, tumor necrosis factor-α, monocyte chemoattractant protein-1, and malondialdehyde increased(P < 0.05), and the activity of superoxide dismutase decreased(P < 0.05).(2)Compared with the preeclampsia group, the cell proliferation and adhesion of the magnesium ion intervention groups significantly increased(P < 0.05); the apoptotic rate decreased(P < 0.05); the expression of Bax and Active Caspase-3 decreased, and the expression of Bcl-2 increased(P < 0.05); the levels of interleukin-6, interleukin-8, tumor necrosis factor-α, monocyte chemoattractant protein-1, malondialdehyde decreased(P < 0.05), and the level of superoxide dismutase increased(P < 0.05). And the improvement in the high concentration magnesium ion group was better than that in the low and middle concentration magnesium ion groups(P < 0.05). To conclude, magnesium ions can promote the proliferation and adhesion of endothelial progenitor cells in vitro, inhibit the secretion of inflammatory factors, and reduce the apoptotic rate and oxidative stress level in patients with preeclampsia.
BACKGROUND: Magnesium ions can reduce the sensitivity of vascular endothelial cells to adverse stimuli and regulate vascular endothelial function by scavenging free radicals, stabilizing cell membranes and inhibiting inflammation. However, the effect of magnesium ions on endothelial progenitor cells is still unclear.OBJECTIVE: To explore the effects of magnesium on the proliferation, adhesion and apoptosis of endothelial progenitor cells in patients with preeclampsia.METHODS: From March 2016 to October 2018, 17 preeclampsia pregnant patients and 15 normal pregnant women who were admitted in the Obstetrics Department of Hebei General Hospital were enrolled. Endothelial progenitor cells were isolated and cultured from umbilical cord blood in the two groups of pregnant women. Endothelial progenitor cells isolated from each preeclampsia pregnant patient were divided into four groups: preeclampsia group was intervened with PBS, and low, middle and high concentration magnesium ion groups were intervened with the medium containing 4, 8, and 16 mmol/L magnesium ions, respectively. Endothelial progenitor cells from normal pregnant women were treated with PBS as control group. After 24 hours of intervention, cell proliferation, adhesion, apoptosis, inflammation(interleukin-6,interleukin-8, tumor necrosis factor-α, monocyte chemoattractant protein-1 levels) and oxidative stress(superoxide dismutase and malondialdehyde levels) were detected. The study protocol was approved by the Ethics Committee of Hebei General Hospital with the approval No. 2013 linshen13.RESULTS AND CONCLUSION:(1) Compared with the control group, the cell proliferation and adhesion of the preeclampsia group were significantly decreased(P < 0.05); the apoptotic rate significantly increased(P < 0.05); the expression of Bax and Active Caspase-3 increased,and the expression of Bcl-2 decreased(both P < 0.05); the levels of interleukin-6, interleukin-8, tumor necrosis factor-α, monocyte chemoattractant protein-1, and malondialdehyde increased(P < 0.05), and the activity of superoxide dismutase decreased(P < 0.05).(2)Compared with the preeclampsia group, the cell proliferation and adhesion of the magnesium ion intervention groups significantly increased(P < 0.05); the apoptotic rate decreased(P < 0.05); the expression of Bax and Active Caspase-3 decreased, and the expression of Bcl-2 increased(P < 0.05); the levels of interleukin-6, interleukin-8, tumor necrosis factor-α, monocyte chemoattractant protein-1, malondialdehyde decreased(P < 0.05), and the level of superoxide dismutase increased(P < 0.05). And the improvement in the high concentration magnesium ion group was better than that in the low and middle concentration magnesium ion groups(P < 0.05). To conclude, magnesium ions can promote the proliferation and adhesion of endothelial progenitor cells in vitro, inhibit the secretion of inflammatory factors, and reduce the apoptotic rate and oxidative stress level in patients with preeclampsia.
引文
[1]Boucas AP, de Souza BM, Bauer AC, et al.Role of Innate Immunity in Preeclampsia:A Systematic Review. Reprod Sci.2017;24(10):1362-1370.
[2]Dhariwal NK,Lynde GC.Update in the Management of Patients with Preeclampsia.Anesthesiol Clin.2017;35(1):95-106.
[3]De Silva DA,Proctor L,von Dadelszen P,et al.Determinants of magnesium sulphate use in women hospitalized at <29 weeks with severe or non-severe pre-eclampsia.PLoS One.2017;12(12):e0189966.
[4]刁桂杰,林永富,吴德荣,等.孕期补镁与妊娠期高血压疾病发生的关系[J].牡丹江医学院学报,2008, 29(4):30-32.
[5]Deng Y,Wang J,He G,et al.Mobilization of endothelial progenitor cell in patients with acute ischemic stroke.Neurol Sci.2018;39(3):437-443.
[6]Ge Q,Zhang H,Hou J,et al.VEGF secreted by mesenchymal stem cells mediates the differentiation of endothelial progenitor cells into endothelial cells via paracrine mechanisms.Mol Med Rep. 2018;17(1):1667-1675.
[7]Wang H,Cheng H,Tang X,et al.The synergistic effect of bone forming peptide-1 and endothelial progenitor cells to promote vascularization of tissue engineered bone.J Biomed Mater Res A. 2018;106(4):1008-1021.
[8]Brodowski L,Burlakov J,Hass S,et al.Impaired functional capacity of fetal endothelial cells in preeclampsia.PloS One.2017;12(5):e0178340.
[9]Gumina DL,Black CP,Balasubramaniam V,et al.Umbilical Cord Blood Circulating Progenitor Cells and Endothelial Colony-Forming Cells Are Decreased in Preeclampsia.Reprod Sci.2017;24(7):1088-1096.
[10]周燕,邹丽,朱剑文,等.子痫前期患者血管内皮祖细胞数量及生物学功能的变化[J].现代妇产科进展, 2009,18(3):208-211.
[11] Katakawa M,Fukuda N,Tsunemi A,et al.Taurine and magnesium supplementation enhances the function of endothelial progenitor cells through antioxidation in healthy men and spontaneously hypertensive rats. Hypertens Res.2016;39(12):848-856.
[12] Almousa LA,Salter AM,Langley-Evans SC.Magnesium deficiency heightens lipopolysaccharide-induced inflammation and enhances monocyte adhesion in human umbilical vein endothelial cells.Magnes Res.2018;31(2):39-48.
[13] Pan S, An L, Meng X, et al. MgCl2 and ZnCl2 promote human umbilical vein endothelial cell migration and invasion and stimulate epithelial-mesenchymal transition via the Wnt/beta-catenin pathway.Exp Ther Med.2017;14(5):4663-4670.
[14] Geng J, Wang L, Qu M, et al. Endothelial progenitor cells transplantation attenuated blood-brain barrier damage after ischemia in diabetic mice via HIF-1alpha.Stem Cell Res Ther.2017;8(1):163.
[15] Li X,Jiang C,Zhao J.Human endothelial progenitor cells-derived exosomes accelerate cutaneous wound healing in diabetic rats by promoting endothelial function. J Diabetes Complications.2016;30(6):986-992.
[16] Li X,Chen C,Wei L,et al.Exosomes derived from endothelial progenitor cells attenuate vascular repair and accelerate reendothelialization by enhancing endothelial function.Cytotherapy.2016;18(2):253-262.
[17]李健,董果雄,张社华.镁对血管内皮细胞缺氧再复氧损伤的保护作用[J].中国微循环,2003,7(4):227-229.
[18] Luo Q,Zeng J,Li W,et al.Silencing of miR155 suppresses inflammatory responses in psoriasis through inflammasome NLRP3 regulation.Int J Mol Med.2018;42(2):1086-1095.
[19] Li BH, Yang K, Wang X.Biodegradable magnesium wire promotes regeneration of compressed sciatic nerves.Neural Regen Res. 2016;11(12):2012-2017.
[20] Tangvoraphonkchai K, Davenport A. Magnesium and Cardiovascular Disease. Adv Chronic Kidney Dis.2018; 25(3):251-260.
[21] Hu T,Xu H, Wang C,et al.Magnesium enhances the chondrogenic differentiation of mesenchymal stem cells by inhibiting activated macrophage-induced inflammation.Sci Rep.2018;8(1):3406.
[22] Chollat C, Marret S.Magnesium sulfate and fetal neuroprotection:overview of clinical evidence.Neural Regen Res. 2018;13(12):2044-2049.
[23]孙海英,张志明,王琰.子痫前期与患者血清镁含量的病例对照研究[J].山西医科大学学报,2015,46(7):658-659.
[24] Colunga T,Dalton S.Building Blood Vessels with Vascular Progenitor Cells. Trends Mol Med. 2018;24(7):630-641.
[25] Yuan F, Chang S, Luo L, et al. cxcl12 gene engineered endothelial progenitor cells further improve the functions of oligodendrocyte precursor cells.Exp Cell Res.2018;367(2):222-231.
[26] Zhao B,Zhao Z,Sun X,et al.Effect of micro strain stress on proliferation of endothelial progenitor cells in vitro by the MAPK-ERK1/2 signaling pathway. Biochem Biophys Res Commun.2017;492(2):206-211.
[27] Liu X,Luo Q,Zheng Y, et al. Notch1 Impairs Endothelial Progenitor Cell Bioactivity in Preeclampsia. Reprod Sci. 2017;24(1):47-56.
[28] Ferguson KK,Meeker JD, McElrath TF, et al. Repeated measures of inflammation and oxidative stress biomarkers in preeclamptic and normotensive pregnancies. Am J Obstet Gynecol.2017;216(5):527.e1-527.e9.
[29]郑浩,李占鲁,沈啸华,等.白藜芦醇抑制叔丁基过氧化物诱导的外周血内皮祖细胞损伤[J].中国病理生理杂志, 2017,33(6):1073-1079.
[30]潘双.镁离子与锌离子对人血管内皮细胞和平滑肌细胞迁移功能的影响[J].辽宁医学院学报,2017,38(2):10-12.
[31]商晓盼,李涛,陆雨桐,等.镁离子在骨代谢中的分子机制研究与进展[J].中国组织工程研究,2016,20(48):7280-7287.
[32] Su NY,Peng TC,Tsai PS,et al.Phosphoinositide 3-kinase/Akt pathway is involved in mediating the anti-inflammation effects of magnesium sulfate.J Surg Res.2013;185(2):726-732.
[2]Dhariwal NK,Lynde GC.Update in the Management of Patients with Preeclampsia.Anesthesiol Clin.2017;35(1):95-106.
[3]De Silva DA,Proctor L,von Dadelszen P,et al.Determinants of magnesium sulphate use in women hospitalized at <29 weeks with severe or non-severe pre-eclampsia.PLoS One.2017;12(12):e0189966.
[4]刁桂杰,林永富,吴德荣,等.孕期补镁与妊娠期高血压疾病发生的关系[J].牡丹江医学院学报,2008, 29(4):30-32.
[5]Deng Y,Wang J,He G,et al.Mobilization of endothelial progenitor cell in patients with acute ischemic stroke.Neurol Sci.2018;39(3):437-443.
[6]Ge Q,Zhang H,Hou J,et al.VEGF secreted by mesenchymal stem cells mediates the differentiation of endothelial progenitor cells into endothelial cells via paracrine mechanisms.Mol Med Rep. 2018;17(1):1667-1675.
[7]Wang H,Cheng H,Tang X,et al.The synergistic effect of bone forming peptide-1 and endothelial progenitor cells to promote vascularization of tissue engineered bone.J Biomed Mater Res A. 2018;106(4):1008-1021.
[8]Brodowski L,Burlakov J,Hass S,et al.Impaired functional capacity of fetal endothelial cells in preeclampsia.PloS One.2017;12(5):e0178340.
[9]Gumina DL,Black CP,Balasubramaniam V,et al.Umbilical Cord Blood Circulating Progenitor Cells and Endothelial Colony-Forming Cells Are Decreased in Preeclampsia.Reprod Sci.2017;24(7):1088-1096.
[10]周燕,邹丽,朱剑文,等.子痫前期患者血管内皮祖细胞数量及生物学功能的变化[J].现代妇产科进展, 2009,18(3):208-211.
[11] Katakawa M,Fukuda N,Tsunemi A,et al.Taurine and magnesium supplementation enhances the function of endothelial progenitor cells through antioxidation in healthy men and spontaneously hypertensive rats. Hypertens Res.2016;39(12):848-856.
[12] Almousa LA,Salter AM,Langley-Evans SC.Magnesium deficiency heightens lipopolysaccharide-induced inflammation and enhances monocyte adhesion in human umbilical vein endothelial cells.Magnes Res.2018;31(2):39-48.
[13] Pan S, An L, Meng X, et al. MgCl2 and ZnCl2 promote human umbilical vein endothelial cell migration and invasion and stimulate epithelial-mesenchymal transition via the Wnt/beta-catenin pathway.Exp Ther Med.2017;14(5):4663-4670.
[14] Geng J, Wang L, Qu M, et al. Endothelial progenitor cells transplantation attenuated blood-brain barrier damage after ischemia in diabetic mice via HIF-1alpha.Stem Cell Res Ther.2017;8(1):163.
[15] Li X,Jiang C,Zhao J.Human endothelial progenitor cells-derived exosomes accelerate cutaneous wound healing in diabetic rats by promoting endothelial function. J Diabetes Complications.2016;30(6):986-992.
[16] Li X,Chen C,Wei L,et al.Exosomes derived from endothelial progenitor cells attenuate vascular repair and accelerate reendothelialization by enhancing endothelial function.Cytotherapy.2016;18(2):253-262.
[17]李健,董果雄,张社华.镁对血管内皮细胞缺氧再复氧损伤的保护作用[J].中国微循环,2003,7(4):227-229.
[18] Luo Q,Zeng J,Li W,et al.Silencing of miR155 suppresses inflammatory responses in psoriasis through inflammasome NLRP3 regulation.Int J Mol Med.2018;42(2):1086-1095.
[19] Li BH, Yang K, Wang X.Biodegradable magnesium wire promotes regeneration of compressed sciatic nerves.Neural Regen Res. 2016;11(12):2012-2017.
[20] Tangvoraphonkchai K, Davenport A. Magnesium and Cardiovascular Disease. Adv Chronic Kidney Dis.2018; 25(3):251-260.
[21] Hu T,Xu H, Wang C,et al.Magnesium enhances the chondrogenic differentiation of mesenchymal stem cells by inhibiting activated macrophage-induced inflammation.Sci Rep.2018;8(1):3406.
[22] Chollat C, Marret S.Magnesium sulfate and fetal neuroprotection:overview of clinical evidence.Neural Regen Res. 2018;13(12):2044-2049.
[23]孙海英,张志明,王琰.子痫前期与患者血清镁含量的病例对照研究[J].山西医科大学学报,2015,46(7):658-659.
[24] Colunga T,Dalton S.Building Blood Vessels with Vascular Progenitor Cells. Trends Mol Med. 2018;24(7):630-641.
[25] Yuan F, Chang S, Luo L, et al. cxcl12 gene engineered endothelial progenitor cells further improve the functions of oligodendrocyte precursor cells.Exp Cell Res.2018;367(2):222-231.
[26] Zhao B,Zhao Z,Sun X,et al.Effect of micro strain stress on proliferation of endothelial progenitor cells in vitro by the MAPK-ERK1/2 signaling pathway. Biochem Biophys Res Commun.2017;492(2):206-211.
[27] Liu X,Luo Q,Zheng Y, et al. Notch1 Impairs Endothelial Progenitor Cell Bioactivity in Preeclampsia. Reprod Sci. 2017;24(1):47-56.
[28] Ferguson KK,Meeker JD, McElrath TF, et al. Repeated measures of inflammation and oxidative stress biomarkers in preeclamptic and normotensive pregnancies. Am J Obstet Gynecol.2017;216(5):527.e1-527.e9.
[29]郑浩,李占鲁,沈啸华,等.白藜芦醇抑制叔丁基过氧化物诱导的外周血内皮祖细胞损伤[J].中国病理生理杂志, 2017,33(6):1073-1079.
[30]潘双.镁离子与锌离子对人血管内皮细胞和平滑肌细胞迁移功能的影响[J].辽宁医学院学报,2017,38(2):10-12.
[31]商晓盼,李涛,陆雨桐,等.镁离子在骨代谢中的分子机制研究与进展[J].中国组织工程研究,2016,20(48):7280-7287.
[32] Su NY,Peng TC,Tsai PS,et al.Phosphoinositide 3-kinase/Akt pathway is involved in mediating the anti-inflammation effects of magnesium sulfate.J Surg Res.2013;185(2):726-732.