同型半胱氨酸对人冠状动脉内皮细胞中组织蛋白酶V和炎症因子表达的影响机制

Mechanism of homocysteine on expression of cathepsin V and inflammatory factors in human coronary artery endothelial cells

  • 摘要:
      目的  分析体外不同浓度同型半胱氨酸(Hcy)介导蛋白激酶C(PKC)/丝裂原活化蛋白激酶(MAPK)/信号转导及转录激活因子1(STAT1)信号通路对人冠状动脉内皮细胞(HCAECs)中组织蛋白酶V(CTSV)和炎症因子表达的影响机制。
      方法  实验分为4组,即对照组(生理盐水)、低浓度组(Hcy 1 mmol/L)、中浓度组(Hcy 5 mmol/L)和高浓度组(Hcy 10 mmol/L), 4组不同溶液分别与HCAECs共培养72 h。采用四氮唑蓝盐化合物(MTS)法检测细胞活力,采用蛋白质印迹法(Western blot)检测PKC、MAPK、STAT1和CTSV相对表达量,采用酶联免疫吸附试验(ELISA)法检测炎症因子血管内皮细胞生长因子受体-1(VEGFR-1)、肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)表达水平。
      结果  与对照组、低浓度组相比,中浓度组、高浓度组细胞活力升高, PKC、MAPK、STAT1和CTSV相对表达量增加, VEGFR-1、TNF-α和IL-1β表达水平升高,差异有统计学意义(P<0.05); 高浓度组PKC、MAPK、STAT1、CTSV相对表达量和VEGFR-1、TNF-α、IL-1β表达水平及细胞活力均高于中浓度组,差异有统计学意义(P<0.05); 对照组与低浓度组各指标比较,差异无统计学意义(P>0.05)。
      结论  Hcy水平升高可激活HCAECs中PKC/MAPK/STAT1信号通路影响CTSV和炎症因子表达,且呈浓度依赖性。Hcy高表达与川崎病导致冠状动脉损伤有关,为疾病的临床干预提供了重要靶点。

     

    Abstract:
      Objective  To analyze the influences of different concentrations of homocysteine (Hcy) mediated protein kinase C (PKC)/mitogen-activated protein kinase (MAPK)/signal transducer in vitro and activator of transcription 1 (STAT1) signaling pathway on the expression of cathepsin V (CTSV) and inflammatory factors in human coronary endothelial cells (HCAECs).
      Methods  The experiment was divided into four groupscontrol group (normal saline), low-concentration (Hcy 1 mmol/L), medium-concentration (5 mmol/L) and high-concentration (10 mmol/L) groups. They were co-cultured with HCAECs at four concentrations for 72 hours. The cell viability was detected by May-Thurner syndrome (MTS), the relative expression of PKC, MAPK, STAT1 and CTSV were detected by western blot, and inflammatory factors including vascular endothelial growth factor receptor-1 (VEGFR-1), tumor necrosis factor-α(TNF-α) and interleukin-1 β (IL-1 β) were detected by enzyme-linked immunosorbent assay.
      Results  Compared with the control group and low-concentration group, the cell viability was increased, relative expression of PKC, MAPK, STAT1 and CTSV were increased, the expression levels of VEGFR-1, TNF- α and IL-1 β were significantly increased in the medium and high-concentration group(P<0.05). The relative expression of PKC, MAPK, STAT1 and CTSV, the expression levels of VEGFR-1, TNF-α and IL-1 β and the cell viability in the high-concentration group were significantly higher than those in the medium-concentration group (P<0.05), while there were no differences between the control group and the low-concentration group in above indicators (P>0.05).
      Conclusion  The increase of Hcy concentration can activate PKC/MAPK/STAT1 signal pathway in HCAECs and affect the expression of CTSV and inflammatory factors with a concentration-dependent manner. High expression of Hcy is closely related to coronary artery injury caused by Kawasaki disease, which provides an important target for clinical intervention.

     

/

返回文章
返回