黄芪甲苷与阿魏酸合用调控JAK-STAT通路促进血管内皮细胞增殖作用的研究
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摘要
目的探讨黄芪甲苷与阿魏酸合用调控JAK-STAT通路对氯化钴所致的缺氧人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVECs)增殖的作用及机制。方法建立人脐静脉内皮细胞氯化钴缺氧损伤模型,采用CCK-8和活细胞染色技术,检测药物对细胞的增殖作用;采用荧光染色技术,观察细胞形态的变化;采用划痕实验测定细胞迁移能力;通过成管实验,观察药物对内皮细胞体外成管作用;采用Western blot法检测各给药组对p-JAK2和p-STAT3蛋白表达的影响。结果与模型组相比,黄芪甲苷与阿魏酸联合作用于氯化钴所致的缺氧HUVECs 24 h,可明显促进HUVECs的增殖,并改善由于缺氧所致的细胞形态的变化,促进细胞的迁移,有利于血管生成。同时可上调p-STAT3和p-JAK2蛋白的表达。结论黄芪甲苷与阿魏酸合用对氯化钴所致的缺氧HUVECs有保护作用,其机制可能与激活JAK-STAT信号通路有关。
Objective To investigate the effect of astragaloside Ⅳ in combination with ferulic acid on the proliferation of cobalt chloride-induced hypoxic human umbilical vein endothelial cells( HUVECs) and its mechanism.Methods A hypoxic injury model of HUVECs was induced by cobalt chloride. CCK-8 and living cell staining techniques were used to detect cell proliferation,fluorescence staining was used to observe the morphological changes of cells,a scratch test was used to determine the migration ability of the cells,and a tube test was used to observe the effect of drugs on endothelial cells. Western blot was used to detect the expression of p-JAK2 and p-STAT3 proteins. Results Compared with the model group,the combination of astragaloside Ⅳ and ferulic acid significantly promoted the proliferation ofHUVECs induced by cobalt chloride for 24 hours,improved morphological changes in cells induced by hypoxia,and promoted cell migration and angiogenesis. Concurrently,the protein expressions of p-STAT3 and p-JAK2 were upregulated.Conclusions Astragaloside Ⅳ combined with ferulic acid protects against cobalt chloride-induced hypoxia in HUVECs and promotes angiogenesis. The mechanism may be related to activating the JAK-STAT signaling pathway.
Objective To investigate the effect of astragaloside Ⅳ in combination with ferulic acid on the proliferation of cobalt chloride-induced hypoxic human umbilical vein endothelial cells( HUVECs) and its mechanism.Methods A hypoxic injury model of HUVECs was induced by cobalt chloride. CCK-8 and living cell staining techniques were used to detect cell proliferation,fluorescence staining was used to observe the morphological changes of cells,a scratch test was used to determine the migration ability of the cells,and a tube test was used to observe the effect of drugs on endothelial cells. Western blot was used to detect the expression of p-JAK2 and p-STAT3 proteins. Results Compared with the model group,the combination of astragaloside Ⅳ and ferulic acid significantly promoted the proliferation ofHUVECs induced by cobalt chloride for 24 hours,improved morphological changes in cells induced by hypoxia,and promoted cell migration and angiogenesis. Concurrently,the protein expressions of p-STAT3 and p-JAK2 were upregulated.Conclusions Astragaloside Ⅳ combined with ferulic acid protects against cobalt chloride-induced hypoxia in HUVECs and promotes angiogenesis. The mechanism may be related to activating the JAK-STAT signaling pathway.
引文
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[12]石定,余州,宋雅,等.阿魏酸在缺氧条件下促进人脐静脉内皮细胞血管形成的实验研究[J].现代生物医学进展,2016,13(16):2452-2455.
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[14] Chen Z,Han ZC. STAT3:a critical transcription activator in angiogenesis[J]. Med Res Rev,2010,28(2):185-200.
[15] Bartoli M,Platt D, Lemtalsi T, et al. VEGF differentially activates STAT3 in microvascular endothelial cells[J]. FASEB J,2003,17(11):1562-1564.
[16] Seu M,Pollock NI,Black J,et al. JAK kinase inhibition abrogates STAT3 activation and head and neck squamous cell carcinoma tumor growth[J]. Neopasia,2015,17(3):256-264.
[17] Zhang X,Tang J,Zhi X,et al. miR-874 functions as a tumor suppressor by inhibiting angiogenesis through STAT3/VEGF-A pathway in gastric cancer[J]. Oucotarget,2015,6(3):1605-1617.
[18] Zhao M,Gao FH,Wang JY,et al. JAK2/STAT3 signaling pathway activation mediates tumor angiogenesis by upregulation of VEGF and b FGF in non-small-cell lung cancer[J]. 2011,73(3):366-374.
[19] Dal Monte M,Martini D,Ristori C,et al. Hypoxia effects on proangiogenic factors in human umbilical vein endothelial cells:functional role of the peptide somatostatin[J]. Naunyn Schmiedebergs Arch Pharmacol,2011,383(6):593-612.
[20] Albasanz-Puig A,Murray J,Namekata M,et al. Opposing roles of STAT-1 and STAT-3 in regulating vascular endothelial growth factor expression in vascular smooth muscle cells[J]. Biochem Biophys Res Commun,2012,428(1):179-184.
[2] Ren X,Han Y,Wang J,et al. An aligned porous electrospun fibrous membrane with controlled drug delivery—An efficient strategy to accelerate diabetic wound healing with improved angiogenesis[J]. Acta Biomater,2018,70:140-153.
[3]李玉梅,杨辛欣,韩旭,等.川芎嗪与黄芪甲苷配伍对人脐静脉内皮细胞血管生成的作用及机制探讨[J].中草药,2017,48(4):722-727.
[4]李玉梅,杨辛欣,韩旭,等.川芎嗪与黄芪甲苷配伍对过氧化氢诱导人脐静脉内皮细胞氧化损伤的保护作用及机制[J].中药新药与临床药理,2017,28(1):18-21.
[5] Li H,Chang J. Stimulation of proangiogenesis by calcium silicate bioactive ceramic[J]. Acta Biomater,2013,9(2):5379-5389.
[6] Bergers G,Benjami LE. Tumorigenesis and the angiogenic switch[J]. Nat Rev Cancer,2003,3:401-410.
[7] Cesare M. Ferulic acid:pharmacological and toxicological aspects[J]. Food and Chemical Toxicology,2014,65:185-195.
[8] Barone E, Calabrese V, Mancuso C. Ferulic acid and its therapeutic potential as a hormetin for age-related diseases[J].Biogerontology,2009,10:97-108.
[9]张娟,金青哲,王兴国.阿魏酸及其衍生物合成及药理研究进展[J].粮油与米脂,2008,19(8):1839-1841.
[10] Lin CM, Chiu JH, Wu IH, et al. Ferulic acid augments angiogenesis via VEGF, PDGF and HIF-1α[J]. J Nutr Biochem,2010,21(7):627-633.
[11]闫盼盼,闫国立,詹向红,等.不同浓度和比例的黄芪甲苷和阿魏酸对人脐静脉内皮细胞增殖的影响[J].中华中医药学刊,2013,31(2):263-264.
[12]石定,余州,宋雅,等.阿魏酸在缺氧条件下促进人脐静脉内皮细胞血管形成的实验研究[J].现代生物医学进展,2016,13(16):2452-2455.
[13] Kiu H,Nicholoson K. Biology and significance of the JAK-STAT signaling pathways[J]. Growth Factors,2012,30(2):88-106.
[14] Chen Z,Han ZC. STAT3:a critical transcription activator in angiogenesis[J]. Med Res Rev,2010,28(2):185-200.
[15] Bartoli M,Platt D, Lemtalsi T, et al. VEGF differentially activates STAT3 in microvascular endothelial cells[J]. FASEB J,2003,17(11):1562-1564.
[16] Seu M,Pollock NI,Black J,et al. JAK kinase inhibition abrogates STAT3 activation and head and neck squamous cell carcinoma tumor growth[J]. Neopasia,2015,17(3):256-264.
[17] Zhang X,Tang J,Zhi X,et al. miR-874 functions as a tumor suppressor by inhibiting angiogenesis through STAT3/VEGF-A pathway in gastric cancer[J]. Oucotarget,2015,6(3):1605-1617.
[18] Zhao M,Gao FH,Wang JY,et al. JAK2/STAT3 signaling pathway activation mediates tumor angiogenesis by upregulation of VEGF and b FGF in non-small-cell lung cancer[J]. 2011,73(3):366-374.
[19] Dal Monte M,Martini D,Ristori C,et al. Hypoxia effects on proangiogenic factors in human umbilical vein endothelial cells:functional role of the peptide somatostatin[J]. Naunyn Schmiedebergs Arch Pharmacol,2011,383(6):593-612.
[20] Albasanz-Puig A,Murray J,Namekata M,et al. Opposing roles of STAT-1 and STAT-3 in regulating vascular endothelial growth factor expression in vascular smooth muscle cells[J]. Biochem Biophys Res Commun,2012,428(1):179-184.