凉血通瘀方对血管内皮细胞增殖、分泌及相关信号通路蛋白表达的影响

Influence of Liangxue Tongyu Fang on Proliferation, Secretion and Related Signaling Pathway Proteins Expression in VEC

  • 摘要: 目的 通过研究凉血通瘀方对血管内皮细胞(VEC)增殖、分泌及相关信号通路蛋白表达的影响,探讨其治疗出血性中风的作用机理。方法 体外培养血管内皮细胞,1 mmol/L过氧化氢(H2O2)造成血管内皮细胞氧化损伤模型;MTT法检测细胞增殖活性,倒置显微镜观察细胞形态,Hochst法观察细胞凋亡,化学法检测细胞上清NO、TNOS和iNOS的表达,Western blot法检测细胞NF-κB、P53、caspase-3和P21蛋白的表达。结果 与H2O2模型组相比较,高、中、低剂量组的凉血通瘀方能明显提高VEC的增殖活力,显著增加细胞上清液中NO、TNOS和iNOS的表达水平;中剂量组凉血通瘀方能改善细胞形态,减少细胞凋亡,降低NF-κB、P53、caspase-3和P21蛋白的表达。结论 凉血通瘀方通过促进细胞增殖、抑制细胞凋亡、提高细胞NO的分泌和TNOS、iNOS的活性,降低NF-κB、P53、caspase-3和P21信号通路蛋白的表达,从而保护血管内皮细胞是凉血通瘀方治疗出血性中风的作用机理。

     

    Abstract: OBJECTIVE By studying the effects of Liangxue Tongyu Fang on proliferation, secretionand and related signaling pathway proteins expression in VEC, to discuss the mechanism of the hemorrhagic stroke. METHODS VEC were cultured in vitro ,H2O2 at 1 mmol/L was used to induce oxidative stress injury model in endothelial cells. MTT was used to detect cell proliferation activity,Inverted microscope was used to observe the cell morphology, Hochst staining was used to examine apoptosis. Chemical method were used to detect the levels of NO, TNOS and iNOS in supernatant cells.Western blot method were used to detect the expression of NF-κB, P53, caspase-3 and P21 protein. RESULTS Compared with the H2O2 model group, Liangxue Tongyu Fang at high, medium and low dose groups obviously increased the proliferation activity and elevated levels of NO,TNOS and iNOS in the cell supernatant. The medium dose group improved cell morphology, reduced apoptosis and the expression of NF-κB, P53, caspase-3 and P21 proteins. CONCLUSION By promoting cell proliferation, inhibiting cell apoptosis, increasing the secretion of NO and the activity of TNOS and iNOS, reducing the expression of NF-κB, P53, caspase-3 and P21 signaling pathway proteins, thus protecting vascular endothelial cells is the mechanisms for the treatment of hemorrhagic stroke.

     

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