高级检索
    王盼, 李言, 张永. 法舒地尔对脂多糖诱导的人脐静脉内皮细胞损伤的影响[J]. 蚌埠医科大学学报, 2017, 42(5): 561-565. DOI: 10.13898/j.cnki.issn.1000-2200.2017.05.001
    引用本文: 王盼, 李言, 张永. 法舒地尔对脂多糖诱导的人脐静脉内皮细胞损伤的影响[J]. 蚌埠医科大学学报, 2017, 42(5): 561-565. DOI: 10.13898/j.cnki.issn.1000-2200.2017.05.001
    shu
    WANG Pan, LI Yan, ZHANG Yong. Effect of fasudil on the human umbilical vein endothelial cells injury induced by lipopolysaccharide[J]. Journal of Bengbu Medical University, 2017, 42(5): 561-565. DOI: 10.13898/j.cnki.issn.1000-2200.2017.05.001
    Citation: WANG Pan, LI Yan, ZHANG Yong. Effect of fasudil on the human umbilical vein endothelial cells injury induced by lipopolysaccharide[J]. Journal of Bengbu Medical University, 2017, 42(5): 561-565. DOI: 10.13898/j.cnki.issn.1000-2200.2017.05.001
    shu

    法舒地尔对脂多糖诱导的人脐静脉内皮细胞损伤的影响

    Effect of fasudil on the human umbilical vein endothelial cells injury induced by lipopolysaccharide

      摘要
    • 摘要: 目的：观察法舒地尔对脂多糖（LPS）诱导的人脐静脉内皮细胞（HUVECs）损伤的影响，探讨其作用机制。方法：用0.1%的Ⅰ型胶原酶消化分离HUVECs，加入内皮细胞完全培养基中培养，采用胰蛋白酶进行消化传代培养，倒置显微镜下观察细胞形态并进行免疫组织化学鉴定。将HUVECs随机分为对照组（内皮细胞培养基培养）、LPS组（内皮细胞培养基+0.1 μg/mL LPS）和法舒地尔组（内皮细胞培养基+0.1 μg/mL LPS+3.275 μg/mL法舒地尔），Western blot法检测RhoA、ROCK2、p-ERM蛋白表达水平，实时荧光定量PCR检测RhoA、ROCK2和p-ERM mRNA表达量。结果：原代培养的内皮细胞在接种后24 h后完全贴壁生长，第3天融合呈铺路石样镶嵌排列，免疫组织化学检测可见第Ⅷ因子相关抗原阳性反应，证实为内皮细胞。LPS组RhoA、ROCK2和p-ERM蛋白及基因表达水平均显著高于对照组（P<0.01）；而法舒地尔组ROCK2和p-ERM蛋白及基因表达均低于LPS组（P<0.05~P<0.01），但2组RhoA蛋白和基因表达水平差异均无统计学意义（P>0.05）。结论：Rho/ROCK/ERM通路在LPS诱导的HUVECs细胞骨架改变中起重要作用，法舒地尔可拮抗LPS诱导的骨架蛋白重排等HUVECs损伤，其机制可能与抑制Rho/ROCK信号通路活化有关。

       

      Abstract: Objective: To observe the effects of fasudil on the human umbilical vein endothelial cells(HUVECs) injury induced by lipopolysaccharide(LPS),explore its mechanism of action.Methods: HUVECs were obtained from human umbilical vein,digested with 0.1% type Ⅰ collagenase,and cultivated and passaged in endothelial cell culture medium.The cell morphology was observed under the inverted microscope,and the cells were identified using immunohistochemistry.HUVECs were randomly divided into the control group(cultured with endothelial cell medium),LPS group(cultured with 0.1 μg/mL LPS) and fasudil group(cultured with 3.275 μg/mL fasudil and 0.1 μg/mL LPS).The protein and mRNA levels of RhoA,ROCK2 and p-ERM were detected using Western blot and quantitative real-time PCR,respectively.Results: The primary cells could adhere to the plate after 24 hours of inoculation,and arranged as paving stone on the third day.The HUVECs identified using the immunohistochemistry staining of Ⅷ factor.related antigen.The protein and mRNA levels of RhoA,ROCK2 and p-ERM in LPS group were significantly higher than those in control group(P<0.01).The protein and mRNA levels of ROCK2 and p-ERM in fasudil group were lower than those in LPS group(P<0.01),the differences of the protein and mRNA levels of RhoA between fasudil group and LPS group were not statistically significant(P>0.05).Conclusions: The Rho/ROCK/ERM signaling pathway plays an important role in the cytoskeletal damage in HUVECs induced by LPS.Fasudil can inhibit the LPS-induced cytoskeletal rearrangement injury,the mechanism of which may be related to inhibiting the Rho/ROCK signaling pathway activation.

       

      • HTML全文
      • 参考文献(13)
      • 相关文章
      • 施引文献
    • 资源附件(0)

    /

    返回文章
    返回