瑞芬太尼对过氧化氢处理脐静脉内皮细胞的抗氧化应激作用

发布时间：2018-11-10 10:40

【摘要】：目的以人脐静脉内皮细胞为细胞模型,建立过氧化氢处理的氧化应激模型,研究瑞芬太尼的抗氧化应激保护机制,并确定信号转导通路。方法过氧化氢0.1mol/L孵育原代培养的人脐静脉内皮细胞,建立细胞损伤模型,然后进行瑞芬太尼保护及相关通路研究。实验共分为九组:空白对照组(C组)、过氧化氢组(H1组)、过氧化氢+SP600125组(H2组)、过氧化氢+SB203580组(H3组)、过氧化氢+PD98059组(H4组);过氧化氢+瑞芬太尼组(HR1组)、过氧化氢+瑞芬太尼+SP600125组(HR2组)、过氧化氢+瑞芬太尼+SB203580组(HR3组)、过氧化氢+瑞芬太尼+PD98059组(HR4组)。H1组、H2组、H3组和H4组仅进行MAPK通路阻断实验,HR1组、HR2组、HR3组和HR4组分别加入瑞芬太尼10ng/ml保护1h。随后分别检测超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量及Caspase-3活性,观察瑞芬太尼抗氧化应激作用并初步确定转导通路;利用RT-PCR观察瑞芬太尼10ng/ml处理前后c-Jun的表达水平,确定转导通路的信号分子。结果H1、H2、H3、H4组SOD活性明显低于C组,MDA含量明显高于C组(P0.05);HR1组SOD活性明显高于H1组,MDA含量明显低于H1组(P0.05);HR2组与H2组SOD活性及MDA含量差异无统计学意义;HR3组SOD活性明显高于H3组,MDA含量明显低于H3组(P0.05);HR4组SOD活性明显高于H4组,MDA含量明显低于H4组(P0.05)。H1、H2、H3、H4组Caspase-3活性明显高于C组(P0.05)。H1组和HR1组c-Jun mRNA表达量明显高于C组,且HR1组明显低于H1组(P0.05)。结论瑞芬太尼10ng/ml可激活JNK通路及其下游信号分子c-Jun,上调SOD活性,降低MDA含量,进而起到抗氧化应激作用。
[Abstract]:Objective to establish an oxidative stress model treated with hydrogen peroxide in human umbilical vein endothelial cells (HUVECs), to study the protective mechanism of remifentanil on antioxidant stress and to determine the signal transduction pathway. Methods the primary cultured human umbilical vein endothelial cells (HUVECs) were incubated with hydrogen peroxide 0.1mol/L, and the injury model was established. Remifentanil was used to protect human umbilical vein endothelial cells (HUVECs). The experiment was divided into nine groups: blank control group (C group), hydrogen peroxide group (H1 group), hydrogen peroxide SP600125 group (H2 group), hydrogen peroxide SB203580 group (H3 group) and hydrogen peroxide PD98059 group (H4 group). Remifentanil hydrogen peroxide group (HR1 group), remifentanil hydrogen peroxide SP600125 group (HR2 group), remifentanil hydrogen peroxide SB203580 group (HR3 group), remifentanil hydrogen peroxide PD98059 group (HR4 group). MAPK pathway blockade was performed only in H3 group and H4 group. HR1 group, HR2 group, HR3 group and HR4 group were treated with remifentanil 10ng/ml for 1 h. The activity of superoxide dismutase (SOD), the content of malondialdehyde (MDA) and the activity of Caspase-3 were detected, and the antioxidant stress of remifentanil was observed and the transduction pathway was preliminarily determined. RT-PCR was used to observe the expression of c-Jun before and after remifentanil 10ng/ml treatment to determine the signal molecule of transduction pathway. Results the activity of SOD and the content of MDA in group H _ (1) H _ (2) H _ (3) H _ (4) were significantly lower than those in group C (P 0.05), the activity of SOD in group H _ (1) was significantly higher than that in group H _ (1) and the content of MDA in group H _ (1) was significantly lower (P < 0.05). There was no significant difference in SOD activity and MDA content between HR2 group and H2 group, SOD activity in HR3 group was significantly higher than that in H3 group, and MDA content in HR3 group was significantly lower than that in H3 group (P0.05). The activity of SOD in HR4 group was significantly higher than that in H4 group, and the content of MDA in H4 group was significantly lower than that in H4 group (P0.05). The Caspase-3 activity of H1H2H3H4 group was significantly higher than that of C group (P0.05). The expression of c-Jun mRNA in H1 group and HR1 group was significantly higher than that in C group. HR1 group was significantly lower than H1 group (P0.05). Conclusion remifentanil 10ng/ml can activate the JNK pathway and its downstream signal molecule c-Jun. it can up-regulate the activity of SOD, decrease the content of MDA, and then play the role of antioxidant stress.
【作者单位】： 潍坊市中医院麻醉科;潍坊市中医院泌尿外一科;
【基金】：潍坊市科技局(2015072)
【分类号】：R614
，

本文编号：2322238