發(fā)布時間：2018-11-10 10:40

【摘要】：目的以人臍靜脈內(nèi)皮細(xì)胞為細(xì)胞模型,建立過氧化氫處理的氧化應(yīng)激模型,研究瑞芬太尼的抗氧化應(yīng)激保護(hù)機(jī)制,并確定信號轉(zhuǎn)導(dǎo)通路。方法過氧化氫0.1mol/L孵育原代培養(yǎng)的人臍靜脈內(nèi)皮細(xì)胞,建立細(xì)胞損傷模型,然后進(jìn)行瑞芬太尼保護(hù)及相關(guān)通路研究。實(shí)驗(yàn)共分為九組:空白對照組(C組)、過氧化氫組(H1組)、過氧化氫+SP600125組(H2組)、過氧化氫+SB203580組(H3組)、過氧化氫+PD98059組(H4組);過氧化氫+瑞芬太尼組(HR1組)、過氧化氫+瑞芬太尼+SP600125組(HR2組)、過氧化氫+瑞芬太尼+SB203580組(HR3組)、過氧化氫+瑞芬太尼+PD98059組(HR4組)。H1組、H2組、H3組和H4組僅進(jìn)行MAPK通路阻斷實(shí)驗(yàn),HR1組、HR2組、HR3組和HR4組分別加入瑞芬太尼10ng/ml保護(hù)1h。隨后分別檢測超氧化物歧化酶(SOD)活性、丙二醛(MDA)含量及Caspase-3活性,觀察瑞芬太尼抗氧化應(yīng)激作用并初步確定轉(zhuǎn)導(dǎo)通路;利用RT-PCR觀察瑞芬太尼10ng/ml處理前后c-Jun的表達(dá)水平,確定轉(zhuǎn)導(dǎo)通路的信號分子。結(jié)果H1、H2、H3、H4組SOD活性明顯低于C組,MDA含量明顯高于C組(P0.05);HR1組SOD活性明顯高于H1組,MDA含量明顯低于H1組(P0.05);HR2組與H2組SOD活性及MDA含量差異無統(tǒng)計(jì)學(xué)意義;HR3組SOD活性明顯高于H3組,MDA含量明顯低于H3組(P0.05);HR4組SOD活性明顯高于H4組,MDA含量明顯低于H4組(P0.05)。H1、H2、H3、H4組Caspase-3活性明顯高于C組(P0.05)。H1組和HR1組c-Jun mRNA表達(dá)量明顯高于C組,且HR1組明顯低于H1組(P0.05)。結(jié)論瑞芬太尼10ng/ml可激活JNK通路及其下游信號分子c-Jun,上調(diào)SOD活性,降低MDA含量,進(jìn)而起到抗氧化應(yīng)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.
【作者單位】： 濰坊市中醫(yī)院麻醉科;濰坊市中醫(yī)院泌尿外一科;
【基金】：濰坊市科技局(2015072)
【分類號】：R614
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本文編號：2322238