子痫前期患者血清可通过sFlt-1-CAV1途径增加肾小球内皮细胞通透性
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摘要
目的:探讨子痫前期(PE)血清中可溶性血管内皮生长因子受体1(sFlt-1)对肾小球内皮细胞(HRGECs)通透性的影响机制。方法:收集20例PE与正常孕妇外周血清,ELISA法检测孕妇外周血中sFlt-1含量。利用PE血清及重组sFlt-1(rsFlt-1)蛋白处理HRGECs,细胞接种于transwell小室,体外细胞培养分组:正常孕妇血清处理组(NG组),PE血清处理组(PE组),正常孕妇血清+rsFlt-1处理组,PE+sFlt-1抑制剂处理组,PE血清+CAV1抑制剂处理组。Evens-blue检测HRGECs对大分子蛋白通透性改变;Western blot法检测caveolin-1(CAV1)蛋白表达。结果:PE血清sFlt-1含量明显高于正常对照组;PE血清、rsFlt-1可使HRGECs对大分子蛋白通透性及CAV1蛋白表达增加。sFlt-1抑制剂、CAV1抑制剂可抑制PE血清、rsFlt-1导致的HRGECs对大分子蛋白通透性增加。结论:PE患者血清可通过sFlt-1-CAV1信号通路增加肾小球内皮细胞对大分子蛋白的通透性,CAV1表达异常可能参与蛋白尿的产生。
Objective:To explore the impacts of sFlt-1 in the preeclampsia serum on the permeability of human renal glomerular endothelial cells(HRGECs).Method:The serum of normal pregnant patients and preeclampsia were collected.The level of sFlt-1 was detected by ELISA.Then culture HRGECs in a device called "transwell".Five groups were divided:the first group was treated with the serum of normal pregnant(NG),the second group was treated with the preeclampsia patients serum(PE),the third group was treated with the same normal serum as the first group and also added 5 ng/ml rsFlt-1,the fourth group was treated with the same preeclampsia serum as the second group and also added sFlt-1 inhibitor in a suitable concentration,the fifth group was treated with the same preeclampsia serum as the second group and also added CAV1 inhibitor in a suitable concentration.And then test the Evans-blue conjunct BSA in the lower chamber to reflect the dynamic permeability of HRGEC.Western blot was used to detect the quantity of caveolin-1 expressed by HRGECs after the cells were stimulated by serums of each group for 24 hours.Result:The sFlt-1 level in preeclampsia serum is higher than that in normal pregnant patients.The permeability of HRGECs and CAV1 protein level were significantly enhanced when they were dealt with preeclampsia serum and normal pregnant serum added sFlt-1.The inhibitor of sFlt-1 or CAV1 could reverse them partly(P<0.05).Conclusion:The preeclampsia serum can lead to hyperpermeability of HRGECs through sFlt-1-CAV1 pathway,which may play an important role in proteinuria formation in preeclampsia patients.
Objective:To explore the impacts of sFlt-1 in the preeclampsia serum on the permeability of human renal glomerular endothelial cells(HRGECs).Method:The serum of normal pregnant patients and preeclampsia were collected.The level of sFlt-1 was detected by ELISA.Then culture HRGECs in a device called "transwell".Five groups were divided:the first group was treated with the serum of normal pregnant(NG),the second group was treated with the preeclampsia patients serum(PE),the third group was treated with the same normal serum as the first group and also added 5 ng/ml rsFlt-1,the fourth group was treated with the same preeclampsia serum as the second group and also added sFlt-1 inhibitor in a suitable concentration,the fifth group was treated with the same preeclampsia serum as the second group and also added CAV1 inhibitor in a suitable concentration.And then test the Evans-blue conjunct BSA in the lower chamber to reflect the dynamic permeability of HRGEC.Western blot was used to detect the quantity of caveolin-1 expressed by HRGECs after the cells were stimulated by serums of each group for 24 hours.Result:The sFlt-1 level in preeclampsia serum is higher than that in normal pregnant patients.The permeability of HRGECs and CAV1 protein level were significantly enhanced when they were dealt with preeclampsia serum and normal pregnant serum added sFlt-1.The inhibitor of sFlt-1 or CAV1 could reverse them partly(P<0.05).Conclusion:The preeclampsia serum can lead to hyperpermeability of HRGECs through sFlt-1-CAV1 pathway,which may play an important role in proteinuria formation in preeclampsia patients.
引文
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[15] 靳艳霞,高劲松,胡静,等.sFlt-1/PlGF比值诊断子痫前期的研究进展[J].中华妇产科杂志,2016,51(7):548-550
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[19] Lobysheva I,Rath G,Sekkali B,et al.Moderate caveolin-1 downregulation prevents NADPH oxidase-dependent endothelial nitric oxide synthase uncoupling by angiotensin II in endothelial cells[J].Arterioscler Thromb Vasc Biol,2011,31(9):2098-2105
[20] Li F,Hagaman JR,Kim HS,et al.eNOS deficiency acts through endothelin to aggravate sFlt-1-induced pre-eclampsia-like phenotype[J].JASN,2012,23(4):652-660
[21] Wu FT,Stefanini MO,Mac Gabhann F,et al.A systems biology perspective on sVEGFR1:its biological function,pathogenic role and therapeutic use[J].J Cell Mol Med,2010,14(3):528-552
[2] Sandoval RM,Wagner MC,Patel M,et al.Multiple factors influence glomerular albumin permeability in rats[J].JASN,2012,23(3):447-457
[3] Chen Y,Huang Y,Jiang R,et al.Syncytiotrophoblast-derived microparticle shedding in early-onset and late-onset severe pre-eclampsia[J].Int J Gynaecol Obstet,2012,119(3):234-238
[4] Li Y,Wu Y,Gong X,et al.Low molecular weight heparin decreases the permeability of glomerular endothelial cells when exposed to pre-eclampsia serum in vitro[J].Nephrol,2012,17(8):754-759
[5] Jiang R,Cai J,Zhu Z,et al.Hypoxic trophoblast HMGB1 induces endothelial cell hyperpermeability via the TRL-4/caveolin-1 pathway[J].J Immunol,2014,193(10):5000-5012
[6] Xu Q,Du F,Zhang Y,et al.Preeclampsia serum induces human glomerular vascular endothelial cell hyperpermeability via the HMGB1-Caveolin-1 pathway[J].J Reprod Immunol,2018,129:1-8
[7] 杜菲,徐钦洋,蒋荣珍.MiR-204降调Caveolin-1表达降低HRGECs对大分子蛋白的通透性[J].现代妇产科进展,2017,26(4):246-250
[8] Jiang R,Teng Y,Huang Y,et al.Preeclampsia serum-induced collagen I expression and intracellular calcium levels in arterial smooth muscle cells are mediated by the PLC-gamma1 pathway[J].Exp Mol Med,2014,46:e115
[9] Ahmad S,Hewett PW,Al-Ani B,et al.Autocrine activity of soluble Flt-1 controls endothelial cell function and angiogenesis[J].Vascular Cell,2011,3(1):15
[10] Chen W,Gassner B,Borner S,et al.Atrial natriuretic peptide enhances microvascular albumin permeability by the caveolae-mediated transcellular pathway[J].Cardiovasc Res,2012,93(1):141-151
[11] 周瑾,孙瑜,杨慧霞.可溶性血管内皮生长因子受体-1在重度子痫前期患者胎盘组织中的表达及意义[J].中华围产医学杂志,2012,15(5):291-293
[12] Cui L,Shu C,Liu Z,et al.The expression of serum sEGFR,sFlt-1,sEndoglin and PLGF in preeclampsia[J].Pregn Hyperten,2018,13:127-132
[13] Brownfoot FC,Tong S,Hannan NJ,et al.Effect of sildenafil citrate on circulating levels of sFlt-1 in preeclampsia[J].Pregn Hyperten,2018,13:1-6
[14] 高劲松,沈晶,蒋宇林,等.妊娠中期孕妇血清sFlt-1、PlGF水平及比值变化预测子痫前期发生的价值[J].中华妇产科杂志,2014,49(1):22-25
[15] 靳艳霞,高劲松,胡静,等.sFlt-1/PlGF比值诊断子痫前期的研究进展[J].中华妇产科杂志,2016,51(7):548-550
[16] Palomaki GE,Haddow JE,Haddow HR,et al.Modeling risk for severe adverse outcomes using angiogenic factor measurements in women with suspected preterm preeclampsia[J].Prenatal Diagn,2015,35(4):386-393
[17] Hussein W,Lafayette RA.Renal function in normal and disordered pregnancy[J].Curr Opin Nephrol Hyperten,2014,23(1):46-53
[18] Adamson SL.sFlt-1 in preeclampsia:trophoblast defense against a decidual VEGFA barrage?[J].J Clin Invest,2014,124(11):4690-4692
[19] Lobysheva I,Rath G,Sekkali B,et al.Moderate caveolin-1 downregulation prevents NADPH oxidase-dependent endothelial nitric oxide synthase uncoupling by angiotensin II in endothelial cells[J].Arterioscler Thromb Vasc Biol,2011,31(9):2098-2105
[20] Li F,Hagaman JR,Kim HS,et al.eNOS deficiency acts through endothelin to aggravate sFlt-1-induced pre-eclampsia-like phenotype[J].JASN,2012,23(4):652-660
[21] Wu FT,Stefanini MO,Mac Gabhann F,et al.A systems biology perspective on sVEGFR1:its biological function,pathogenic role and therapeutic use[J].J Cell Mol Med,2010,14(3):528-552