醛糖还原酶抑制剂ARI减轻屋尘螨诱导的人支气管上皮细胞系16-HBE合成iNOS

Aldose Reductase Inhibitor ARI Attenuates House Dust Mite-InducediNOS Synthesis in Human Bronchial Epithelial Cell Line 16-HBE

目的探究醛糖还原酶抑制剂ARI减轻屋尘螨诱导的人支气管上皮细胞系16-HBE合成诱生型一氧化氮合酶(iNOS)。方法将细胞分为对照组,屋尘螨刺激人支气管上皮细胞(模型组),醛糖还原酶抑制剂干预(干预组),地塞米松干预(阳性对照组)。CCK-8法及流式细胞测量术检测细胞增殖及凋亡;硝酸还原酶法测定NO水平和iNOS活性;ELISA检测白介素6(IL-6)和白介素29(IL-29)水平;RT-qPCR检测细胞中诱生型一氧化氮合酶(iNOS)和内皮型一氧化氮合酶(eNOS)mRNA表达;Western blot检测Akt、PI3K、p-Akt、p-PI3K蛋白表达。结果和对照组相比,模型组细胞增殖能力和细胞中eNOS mRNA表达显著降低(P<0.05);和模型组相比,干预组细胞增殖能力和细胞中eNOS mRNA表达显著增加(P<0.05)。和对照组相比,模型组细胞凋亡率、细胞中NO和iNOS表达、IL-6和IL-29水平及Akt、PI3K、p-Akt、p-PI3K蛋白表达均显著增加(P<0.05);和模型组相比,干预组细胞凋亡率、细胞中NO和iNOS表达、IL-6和IL-29水平及Akt、PI3K、p-Akt、p-PI3K蛋白表达均显著降低(P<0.05)。结论醛糖还原酶抑制剂调控人支气管上皮细胞生物学活性,减少支气管哮喘引起的支气管上皮细胞合成iNOS,抑制炎性表达,其作用机制可能与调控PI3K/Akt信号通路有关。

Objective To explore aldose reductase inhibitors to reduce the inducible nitricoxide synthase(iNOS)synthesis of human bronchial epithelial cells(16-HBE)induced by house dust mite.Methods Human bronchial epithelial cells were divided into the control group,house dust mites stimulated human bronchial epithelial cells(model group),aldose reductase inhibitor intervention(intervention group),and dexamethasone intervention(positive control group).CCK-8 and flow cytometry was applied to detect cell proliferation and apoptosis.Nitrate reductase method was used to detect NO level and iNOS activity.ELISA was conducted to detect interleukin 6(IL-6)and interleukin 29(IL-29)content.RT-qPCR was performed to detect cells INOS and eNOS mRNA expressions.Western blot was used to detect of Akt,PI3K,p-Akt,p-PI3K protein expressions.Results Compared with the control group,the cell proliferation ability and eNOS mRNA expression in the model group were significantly reduced(P<0.05);Compared with the model group,the cell proliferation ability and eNOS mRNA expression in the intervention group were significantly increased(P<0.05).Compared with the control group,the apoptosis rate,the expression of NO and iNOS,the content of IL-6 and IL-29,and the protein expression of Akt,PI3K,p-Akt,and p-PI3K in the model group were significantly increased(P<0.05);Compared with the model group,the intervention group’s apoptosis rate,the expression of NO and iNOS,the content of IL-6 and IL-29,and the protein expressions of Akt,PI3K,p-Akt,and p-PI3K in the intervention group were significantly reduced(P<0.05).Conclusion Aldose reductase inhibitor regulates the biological activity of human bronchial epithelial cells,reduces the synthesis of iNOS in bronchial epithelial cells caused by bronchial asthma,and inhibits inflammatory expressions.Its mechanism may be related to the regulation of PI3K/Akt signaling pathway.