高级检索

    黄连碱对人脐静脉内皮细胞内质网应激的影响

    Effect of coptisine on endoplasmic reticulum stress in human umbilical vein endothelial cells

      摘要
    • 摘要:
      目的 建立人脐静脉内皮细胞（HUVECs）内质网应激细胞模型，观察黄连碱对HUVECs以及内质网模型细胞生长活性的影响，对CHOP、GRP78基因以及蛋白表达的影响。
      方法 使用10 μg/mL的衣霉素诱导HUVECs 18 h后，加入不同浓度的黄连碱再处理24 h。CCK8法检测不同浓度黄连碱对HUVECs的活性影响以及黄连碱对衣霉素诱导后的HUVECs的活性影响；采用RT-RCR法比较各组细胞中CHOP、GRP78的mRNA的表达情况；采用Western blotting和细胞免疫荧光法比较各组细胞中CHOP，GRP78的蛋白表达情况。
      结果 随着黄连碱浓度的增加，在1~20 μmol/L范围内的黄连碱对HUVECs的活性无影响，浓度为20 μmol/L黄连碱作用24 h后，对衣霉素诱导的HUVECs活性具有保护作用（P < 0.01）。加入衣霉素诱导18 h，HUVECs中CHOP、GRP78 mRNA水平的表达增高（P < 0.01），再加入黄连碱继续诱导24 h后，黄连碱可以抑制衣霉素所诱导的HUVECs中CHOP、GRP78 mRNA水平的表达增高（P < 0.01）。加入衣霉素诱导18 h，HUVECs中CHOP、GRP78蛋白水平的表达增高（P < 0.01），再加入黄连碱继续诱导24 h后，发现黄连碱可以抑制衣霉素所诱导的HUVECs中CHOP、GRP78蛋白水平的表达增高（P < 0.01），与衣霉素诱导组相比数据具有统计学意义，细胞免疫荧光染色结果与Western blotting结果相似。
      结论 黄连碱可以减轻由衣霉素诱导的HUVECs过度内质网应激反应，并可提高受损细胞的活性；并可通过下调CHOP、GRP78的表达改善HUVECs过度内质网应激反应。

       

      Abstract:
      Objective To endoplasmic reticulum stress cell model of human umbilical vein endothelial cells(HUVECs), to estoblish observe the effects of coptisine on the growth activity of HUVECs cells, and expression levels of CHOP and GRP78 gene and protein.
      Methods HUVECs were induced by tunicamycin(TM) at a concentration of 10 μg/mL for 18 h, and then treated with coptisine at different concentrations for 24 h.The CCK8 was used to detect the effects of different concentrations of coptisine on the activity of HUVECs and activity of HUVECs induced by TM.The mRNA expression levels of CHOP and GRP78 in each group were detected using RT-PCR.The protein expression levels of CHOP and GRP78 in each group were detected using Western blotting and cellular immunofluorescence.
      >Results With the concentration of coptisine increasing, the coptisine in the range of 1-20 μmol/L had no effect on the activity of HUVECs.After 24 hours, the concentration of coptisine in the range of 20 μmol/L played a protective effect on the activity of HUVECs induced by TM(P < 0.01).The mRNA expression levels of CHOP and GRP78 in HUVECS increased after 18 hours of TM induction(P < 0.01), after 24 hours of continuous induction with coptisine, it was found that coptisine could inhibit the mRNA expressions of CHOP and GRP78 in TM-induced HUVECs(P < 0.01).The protein expression levels of CHOP and GRP78 in HUVECs increased after 18 hours of TM induction(P < 0.01), and after 24 hours of continuous induction with coptisine, it was found that coptisine could inhibit the protein expression levels of CHOP and GRP78 in TM-induced HUVECs(P < 0.01).The results of cellular immunofluorescence staining were similar to those of Western blotting.
      Conclusions Coptisine can alleviate the excessive ER stress induced by TM in HUVECs, and improve the activity of damaged cells.Coptisine can improve the excessive ER stress response of HUVECs by down-regulating the expressions of CHOP and GRP78.

       

      • HTML全文
      • 参考文献(16)
      • 相关文章
      • 施引文献
    • 资源附件(0)

    /

    返回文章
    返回