[1]宋振强,王靖宇,周赛君,等.β2-糖蛋白Ⅰ对VEGF诱导的巨噬细胞 迁移的影响[J].国际内分泌代谢杂志,2020,40(05):304-309.[doi:10.3760/cma.j.cn121383-20200425-04072]
　Song Zhenqiang,Wang Jingyu,Zhou Saijun,et al.Effects of β2-glycoprotein Ⅰ on the migration of macrophage induced by VEGF[J].International Journal of Endocrinology and Metabolism,2020,40(05):304-309.[doi:10.3760/cma.j.cn121383-20200425-04072]
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β2-糖蛋白Ⅰ对VEGF诱导的巨噬细胞 迁移的影响()
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《国际内分泌代谢杂志》[ISSN:1673-4157/CN:12-1383/R]

卷:
40
期数:
2020年05期
页码:
304-309
栏目:
论著
出版日期:
2020-09-20

文章信息/Info

Title:
Effects of β2-glycoprotein Ⅰ on the migration of macrophage induced by VEGF
作者:
宋振强; 王靖宇; 周赛君; 于珮
天津医科大学朱宪彝纪念医院,国家卫生健康委员会激素与发育重点实验室,天津市代谢性疾病重点实验室 300070
Author(s):
Song Zhenqiang;  Wang Jingyu;  Zhou Saijun;  Yu Pei
Tianjin Medical University Chu Hsien-Ⅰ Memorial Hospital, NHC Key Laboratory of Hormones and Development(Tianjin Medical University), Tianjin Key Laboratory of Metabolic Diseases, Tianjin 300070, China
关键词:
β2-糖蛋白Ⅰ;  血管内皮生长因子;  巨噬细胞;  糖尿病;  p38丝裂原活化蛋白激酶
Keywords:
β2-glycoprotein Ⅰ;  Vascular endothelial growth factor;  Macrophage;  Diabetes mellitus;  p38 Mitogen-activated protein kinase
DOI:
10.3760/cma.j.cn121383-20200425-04072
文献标志码:
A
摘要:
目的 观察血管内皮生长因子(VEGF)对小鼠单核/巨噬细胞系RAW 264.7的趋化作用,以及β2-糖蛋白Ⅰ(β2-GP)对VEGF诱导巨噬细胞迁移的影响,探讨其可能机制。方法 体外培养RAW 264.7小鼠单核/巨噬细胞系,实验分为5组:空白对照组、VEGF组、VEGF+β2-GP组、VEGF+SB203580(p38抑制剂)组、β2-GP组。Transwell实验观察β2-GP对VEGF诱导巨噬细胞迁移的影响。ELISA法检测β2-GP对VEGF诱导巨噬细胞分泌趋化因子单核细胞趋化蛋白(MCP)-1、白细胞介素(IL)-8以及巨噬细胞炎性蛋白-1β(MIP-1β)的影响。Western印迹检测β2-GP对VEGF干预下巨噬细胞VEGF受体1(VEGFR1)和p38丝裂原活化蛋白激酶(MAPK)信号通路的影响。结果(1)与空白对照组相比,VEGF组巨噬细胞迁移的数量增加(t=22.089,P<0.05),与VEGF组相比,VEGF+β2-GP组和VEGF+SB203580组巨噬细胞迁移明显降低(t=22.687、63.625,P均<0.05)。(2)与空白对照组相比,VEGF组巨噬细胞趋化因子MCP-1、IL-8及MIP-1β的分泌增加(t=3.339、11.140、2.254,P均<0.05),与VEGF组相比,VEGF+β2-GP组和VEGF+SB203580组抑制VEGF诱导的3种趋化因子的分泌减少(t=3.148、2.402、2.798、6.754、4.459、5.528,P均<0.05)。(3)与空白对照组相比,VEGF组巨噬细胞VEGFR1的表达及p38MAPK通路的磷酸化程度增加(t=5.161,P<0.05); 与VEGF组相比,VEGF+β2-GP组巨噬细胞VEGFR1表达及p38MAPK通路的磷酸化程度降低(t=4.965,P<0.05)。结论 VEGF通过促进巨噬细胞VEGFR1表达,活化p38MAPK,促进巨噬细胞的迁移以及趋化因子MCP-1、IL-8、MIP-1β的分泌; β2-GP可抑制VEGFR1的表达,部分抑制p38MAPK磷酸化,从而抑制VEGF诱导的巨噬细胞迁移及趋化因子分泌。
Abstract:
Objective To observe the chemotactic effects of vascular endothelial growth factor(VEGF)on macrophages and the effects of β2-glycoprotein Ⅰ(β2-GP)on the migration of macrophage induced by VEGF through RAW 264.7 incubation, and to discuss the possible mechanisms.Methods RAW 264.7 cells were incubated and divided into five groups: control group,VEGF group,VEGF+β2-GP group, VEGF+SB203580(p38 inhibitor)group, β2-GP group. Transwell cell migration assay was used to observe the chemotaxis of the cells in response to VEGF, and the effect of β2-GP on the macrophage migration induced by VEGF. ELISA was used to detect the excretion of monocyte chemotactic proteins-1(MCP-1), interleukin-8(IL-8)and mitochondrial intermediate peptidase-1β(MIP-1β). Western blotting was used to detect the expression of vascular endothelial growth factor receptor 1(VEGFR1)and activation of p38 mitogen-activated protein kinase(MAPK)signal transduction pathways.Results(1)Compared with control group, the number of macrophage migration in VEGF group increased(t=22.089, P<0.05). Compared with VEGF group, the macrophage migration in VEGF+β2-GP group and VEGF+SB203580 group was significantly reduced(t=22.687, 63.625, all P<0.05).(2)Compared with control group, the secretion of macrophage chemokines MCP-1, IL-8 and MIP-1 increased in VEGF group(t=3.339, 11.140, 2.254, all P<0.05), and the secretion of the three chemokines induced by inhibiting VEGF decreased in VEGF+β2-GP group and VEGF+SB203580 group(t=3.148, 2.402, 2.798, 6.754, 4.459, 5.528, all P<0.05).(3)Compared with control group, VEGFR1 expression and p38MAPK pathway phosphorylation were increased in VEGF group(t=5.161, P<0.05), while VEGFR1 expression and p38MAPK pathway phosphorylation were decreased in VEGF+β2-GP group(t=4.965, P<0.05).Conclusions VEGF can induce the migration of macrophages and secretion of chemokines MCP-1, IL-8 and MIP-1β through the increase of VEGFR1 on the macrophages and activate the p38MAPK transduction pathway; β2-GP can inhibit the expression of VEGFR1 and partially inhibit the phosphorylation of p38MAPK, thereby inhibiting VEGF-induced migration of macrophage and secretion of chemokine.

参考文献/References:

[1] Wang L, Gao P, Zhang M, et al. Prevalence and ethnic pattern of diabetes and prediabetes in China in 2013[J].JAMA,2017,317(24):2515-2523.DOI:10.1001/jama.2017.7596.
[2] American Diabetes Association.Microvascular complications and foot care[J].Diabetes Care, 2017,40(Suppl 1):S88-S98.DOI:10.2337/dc17-S013.
[3] Zhang L, Long J, Jiang W, et al. Trends in chronic kidney disease in China[J].N Engl J Med, 2016, 375(9):905-906.DOI:10.1056/NEJMc1602469.
[4] Wada J, Makino H. Inflammation and the pathogenesis of diabetic nephropathy[J].Clin Sci(Lond),2013, 124(3):139-152. DOI: 10.1042/CS20120198.
[5] Sasaki M, Shikata K, Okada S, et al. The macrophage is a key factor in renal injuries caused by glomerular hyperfiltration[J].Acta Med Okayama,2011,65(2):81-89.DOI:10.1002/emmm.201100126.
[6] Klessens CQF, Zandbergen M, Wolterbeek R, et al. Macrophages in diabetic nephropathy in patients with type 2 diabetes[J].Nephrol Dial Transplant,2017,32(8):1322-1329.DOI:10.1093/ndt/gfw260.
[7] Yu WM, Wang H, Ren XJ, et al. Experimental study of leflunomide on renal protective effect and on inflammatory response of streptozotocin induced diabetic rats[J].Nephrology(Carlton),2012,17(4):380-389.DOI:10.1111/j.1440-1797.2012.01563.x.
[8] Chow FY, Nikolic-Paterson DJ, Atkins RC, et al. Macrophages in streptozotocin-induced diabetic nephropathy:potential role in renal fibrosis[J].Nephrol Dial Transplant,2004,19(12):2987-2996.DOI:10.1093/ndt/gfh441.
[9] Melincovici CS, Bo �瘙 塂 ca AB, �瘙 塁 u �瘙 塂 man S, et al. Vascular endothelial growth factor(VEGF)-key factor in normal and pathological angiogenesis[J].Rom J Morphol Embryol,2018,59(2):455-467.
[10] Zorena K,Kowalewska B,Szmigierokawko M,et al.No effect of yeast-like fungi on lipid metabolism and vascular endothelial growth factor level in children and adolescents with type 1 diabetes mellitus[J].Ital J Pediatr,2016,42(1):107.DOI:10.1186/s13052-016-0317-9.
[11] Bus P,Scharpfenecker M,Van Der Wilk P. The VEGF-A inhibitor sFLT-1 improves renal function by reducing endothelial activation and inflammation in a mouse model of type 1 diabetes[J]. Diabetologia,2017,60(9):1813-1821.DOI:10.1007/s00125-017-4322-3.
[12] Chen HB, Lu JX, Li Q, et al. The protective effect of the RAS inhibitor on diabetic patients with nephropathy in the context of VEGF suppression[J].Acta Pharmacol Sin, 2009, 30(2):242-250.DOI:10.1038/aps.2008.28.
[13] Tchaikovski V, Fellbrich G, Waltenberger J. The molecular basis of VEGFR-1 signal transduction pathways in primary human monocytes[J].Arterioscler Thromb Vasc Biol, 2008, 28(2):322-328.DOI:10.1161/ATVBAHA.107.158022.
[14] Wang T, Chen SS, Chen R, et al. Reduced beta 2 glycoprotein I improve diabetic nephropathy via inhibiting TGF-β1-p38 MAPK pathway[J].Int J Clin Exp Med, 2015,8(5):6852-6865.
[15] Chiu WC, Chiou TJ, Chung MJ, et al. β2-glycoprotein I inhibits vascular endothelial growth factor-induced angiogenesis by suppressing the phosphorylation of extracellular signal-regulated kinase 1/2, Akt, and endothelial nitric oxide synthase[J].PLoS One,2016,11(8):e0161950.DOI:10.1371/journal.pone.0161950.
[16] Wang C, Niu DM, Hu J, et al.Elevated serum β2-glycoprotein-Ⅰ-lipoprotein(a)complexes levels are associated with the presence and complications in type 2 diabetes mellitus[J].Diabetes Res Clin Pract, 2013,100(2):250-256.DOI:10.1016/j.diabres.2013.03.002.

备注/Memo

备注/Memo:
基金项目:国家自然科学基金(30971393) 通信作者:宋振强,Email: Zsong@tmu.edu.cn
更新日期/Last Update: 2020-09-20