Effects of Xingnaojing on recombinant human tumor necrosis factor-mediated tissue-type plasminogen activator and plasminogen activator inhibitor-1 expression in human umbilical cord vein endothelial cells

doi:10.3969/j.issn.2095-4344.2015.11.015

Abstract

Abstract:

BACKGROUND: We have found that Xingnaojing can effectively inhibit mediators of inflammatory mediators and promote fibrinolytic activity in a rabbit model of myocardial ischemia and reperfusion. However, it is not completely clear what is the action of mechanism of fibrinolytic system activity in human umbilical vein endothelial cells.

OBJECTIVE: To investigate the effects of Xingnaojing on recombinant human tumor necrosis factor α-mediated tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1) expression in human umbilical cord vein endothelial cells.

METHODS: The passages 3-5 of human umbilical cord vein endothelial cells were obtained, and were induced by adding 10 μg/L recombinant human tumor necrosis factor α in the culture medium. Xingnaojing in different concentrations (5, 10, 20 mL/L) were injected in the Xingnaojing group, and fluvastatin (1 μmol/L) was added as positive control group. A blank control group of simple human umbilical cord vein endothelial cells culture was set up. After 24 hours of cultivation, the production of t-PA and PAI-1 in the cultured medium was determined by ELISA. The mRNA expression of t-PA and PAI-1 was measured by reverse transcript-PCR.

RESULTS AND CONCLUSION: Compared with the blank control group, the secretion and mRNA expression of PAI-1 in the group treated with recombinant human tumor necrosis factor α (10 μg/L) were significantly increased (P < 0.05); however, the secretion and mRNA expression of t-PA in the group treated with recombinant human tumor necrosis factor α (10 μg/L) were significantly decreased (P < 0.05). The secretion and mRNA expression of t-PA were significantly higher in the Xingnaojing group at different concentrations than in the recombinant human tumor necrosis factor α group (P < 0.05); however, the secretion and mRNA expression of PAI-1 were significantly lower in the former group than the latter group (P < 0.05), which was in a dose-dependent manner. Xingnaojing can effectively improve the fibrinolytic function of human umbilical cord vein endothelial cells mediated by recombinant human tumor necrosis factor α.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: Drugs, Chinese Herbal, Tumor Necrosis Factor-alpha, Tissue Plasminogen Activator

CLC Number:

R318

Ouyang Hai-chun, Wu Wo-dong, Zhong Dong-mei, Wu Yan-xian, Li Wen-jie, Chen Xi-ming. Effects of Xingnaojing on recombinant human tumor necrosis factor-mediated tissue-type plasminogen activator and plasminogen activator inhibitor-1 expression in human umbilical cord vein endothelial cells[J]. Chinese Journal of Tissue Engineering Research, 2015, 19(11): 1717-1721.

Figures/Tables 2

2.2 醒脑静干预下各实验组人脐静脉内皮细胞分泌组织型纤溶酶原激活物和纤溶酶原激活物抑制剂1的变化从表1可以看出，与空白对照组相比，10 mg/L重组人肿瘤坏死因子α可以显著减少人脐静脉内皮细胞组织型纤溶酶原激活物的分泌(P < 0.05)和显著促进人脐静脉内皮细胞纤溶酶原激活物抑制剂1的分泌(P < 0.05)；而各组的醒脑静、1 μmol/L氟伐他汀均可显著增加10 mg/L重组人肿瘤坏死因子α所引起的组织型纤溶酶原激活物的分泌减少(P < 0.05)，均可显著抑制10 mg/L重组人肿瘤坏死因子α所引起的纤溶酶原激活物抑制剂1的分泌增加(P < 0.05)；氟伐他汀对重组人肿瘤坏死因子α介导人脐静脉内皮细胞分泌组织型纤溶酶原激活物和纤溶酶原激活物抑制剂1的影响比醒脑静作用更强，与 5 mL/L及10 mL/L醒脑静比差异有显著性意义(P < 0.05)，而与20 mL/L醒脑静比差异无显著性意义(P > 0.05)。 2.3 各实验组纤溶酶原激活物抑制剂1、组织型纤溶酶原激活物实时荧光定量PCR结果从表2可以看出，与空白对照组相比，10 mg/L重组人肿瘤坏死因子α可以显著减少人脐静脉内皮细胞组织型纤溶酶原激活物的mRNA表达 (P < 0.05)和显著促进人脐静脉内皮细胞纤溶酶原激活物抑制剂1的mRNA表达(P < 0.05)；而各组的醒脑静、1 μmol/L氟伐他汀均可显著增加10 mg/L重组人肿瘤坏死因子α所引起的组织型纤溶酶原激活物的mRNA表达减少(P < 0.05)，均可显著抑制10 mg/L重组人肿瘤坏死因子α所引起的纤溶酶原激活物抑制剂1的mRNA表达增加(P < 0.05)；氟伐他汀对重组人肿瘤坏死因子α介导人脐静脉内皮细胞分泌组织型纤溶酶原激活物和纤溶酶原激活物抑制剂1的mRNA表达影响比各浓度的醒脑静作用更强，与 5 mL/L醒脑静及10 mL/L醒脑静比较差异有显著性意义 (P < 0.05)，而与20 mL/L醒脑静比较差异无显著性意义 (P > 0.05)。"

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