Avezov Katia, Reznick Abraham Z, Aizenbud Dror
Department of Anatomy and Cell Biology, Rappaport Faculty of Medicine, Technion - Israel Institute of Technology, Haifa, Israel; Orthodontic and Craniofacial Department, School of Graduate Dentistry Rambam Health Care Campus, Haifa, Israel.
Department of Anatomy and Cell Biology, Rappaport Faculty of Medicine, Technion - Israel Institute of Technology, Haifa, Israel.
Toxicol In Vitro. 2014 Jun;28(4):485-91. doi: 10.1016/j.tiv.2014.01.004. Epub 2014 Jan 18.
Cigarette smoke (CS) is a significant environmental source of human exposure to chemically active saturated (acetaldehyde) and α,β-unsaturated aldehydes (acrolein) inducing protein carbonylation and dysfunction. The exposure of oral tissues to environmental hazards is immense, especially in smokers. The objectives of the current study were to examine the effect of aldehydes originating from CS on intracellular proteins of oral keratinocytes and to observe the antioxidant response in these cells. Intracellular protein carbonyl modification under CS, acrolein and acetaldehyde exposure in the HaCaT keratinocyte cell line, representing oral keratinocytes was examined by Western blot. Possible intracellular enzymatic dysfunction under the above conditions was examined by lactate dehydrogenase (LDH) activity assay. Oxidative stress response was investigated, by DCF (2,7-dichlorodihydrofluorescein) assay and GSH (glutathione) oxidation. Intracellular protein carbonyls increased 5.2 times after CS exposure and 2.7 times after exposure to 1 μmol of acrolein. DCF assay revealed an increase of fluorescence intensity 3.2 and 3.1 times after CS and acrolein exposure, respectively. CS caused a 72.5% decrease in intracellular GSH levels compared to controls. Activity of intracellular LDH was preserved. α,β-Unsaturated aldehydes from CS are capable of intracellular protein carbonylation and have a role in intracellular oxidative stress elevation in keratinocytes, probably due to the reduction in GSH levels.
香烟烟雾（CS）是人类接触化学活性饱和醛（乙醛）和α,β-不饱和醛（丙烯醛）的一个重要环境来源，这些醛会导致蛋白质羰基化和功能障碍。口腔组织暴露于环境危害中的情况非常严重，尤其是在吸烟者中。本研究的目的是检测源自香烟烟雾的醛对口腔角质形成细胞内蛋白质的影响，并观察这些细胞中的抗氧化反应。通过蛋白质免疫印迹法检测了代表口腔角质形成细胞的HaCaT角质形成细胞系在暴露于香烟烟雾、丙烯醛和乙醛时细胞内蛋白质的羰基修饰情况。通过乳酸脱氢酶（LDH）活性测定检测了上述条件下可能存在的细胞内酶功能障碍。通过DCF（2,7-二氯二氢荧光素）测定和谷胱甘肽（GSH）氧化研究了氧化应激反应。暴露于香烟烟雾后细胞内蛋白质羰基增加了5.2倍，暴露于1μmol丙烯醛后增加了2.7倍。DCF测定显示，暴露于香烟烟雾和丙烯醛后荧光强度分别增加了3.2倍和3.1倍。与对照组相比，香烟烟雾使细胞内谷胱甘肽水平降低了72.5%。细胞内LDH的活性保持不变。香烟烟雾中的α,β-不饱和醛能够引起细胞内蛋白质羰基化，并在角质形成细胞内氧化应激升高过程中起作用，这可能是由于谷胱甘肽水平降低所致。
