血管活性肠肽对兔支气管上皮细胞抗臭氧损伤的保护作用  

CYTOPROTECTIVE EFFECT OF VASOACTIVE INTESTINALPEPTIDE ON RABBIT BRONCHIAL EPITHELIAL CELLS DAMAGED BY OZONE EXPOSURE

作　　者：秦晓群[1] 孙秀泓[1] 张长青[1] 

机构地区：[1]湖南医科大学生理学教研室

出　　处：《中国应用生理学杂志》1998年第3期250-253,共4页Chinese Journal of Applied Physiology

基　　金：国家自然科学基金

摘　　要：用支气管刷洗法收集新西兰兔支气管上皮细胞（ＢＥＣ），以臭氧（Ｏ３）攻击培养的ＢＥＣ，建立细胞损伤模型。测定ＢＥＣ的３Ｈ释放率计算Ｏ３的细胞毒指数（ＣＩ）、测定细胞内丙二醛（ＭＤＡ）的含量反映细胞氧化性损伤的程度，测定细胞内过氧化氢酶（ＣＡＴ）活性及还原型和氧化型谷胱甘肽（ＧＳＨ和ＧＳＳＧ）的含量反映细胞抗氧化能力。观察血管活性肠肽（ＶＩＰ）预处理对ＢＥＣ的细胞保护作用并初步探讨其保护机制。观察到：ＢＥＣ的３Ｈ释放率与Ｏ３暴露时间成正比；Ｏ３暴露２ｈ使ＭＤＡ含量和ＧＳＳＧ含量明显增加，ＧＳＨ减少；ＶＩＰ预处理呈剂量依赖性降低Ｏ３暴露的ＣＩ值、降低ＭＤＡ和ＧＳＳＧ含量、增加ＧＳＨ及ＧＳＨ／ＧＳＳＧ比值、增加ＣＡＴ活性，显示出细胞保护效应；ＶＩＰ的保护效应可被放线菌素Ｄ（Ａ－Ｄ）或蛋白激酶Ｃ阻断剂Ｈ７部分取消。结果表明：Ｏ３暴露会导致ＢＥＣ损伤，ＶＩＰ可通过增强ＢＥＣ的抗氧化能力而保护ＢＥＣ，ＶＩＰ的信号在细胞内的转导途径与基因转录及依赖ＰＫＣ的酶蛋白磷酸化有关。his article established an injury cell model of rabbit bronchial epithelial cells (BEC) exposed to 1.5 ppm ozone (O3). Cells were harvested by the technique of brushing rabbit bronchial and cultured in serumfree RPMI 1640 medium. The 3H released from BEC was determined and the cytotoxic index (CI) of O3 was calculated. The level of lipoperoxidation product malondialdehyde (MDA) in cells reflected the oxidant damage of BEC. For estimating the antioxidant ability of BEC, content of intracellular glutathiones (both GSH and GSSG) and the activity of hydrogen peroxidase (catalase, CAT) in cells were assayed. The results show that O3 exposure induced a timedependent injury(r=0.99) on BEC and brought about an obvious depletion on GSH level (P<0.01). Pretreatment with vasoactive intestinal peptide (VIP), the CI value induced by O3　were dosedependently decreased (r=0.99) and the content of MDA was diminished (P<0.01), which turned out that VIP can protect BEC from injury　under O3 attack, VIP pretreatment elevated the content of GSH (P<0.01), the ratio of GSH/GSSG (P<0.01) and the CAT activity (P<0.01), by which the antioxidant ability of BEC was improved. All of the above mentioned protective effects of VIP could be attenuated by the transeription inhibitor actinomycin D (AD) and the protein kinase C blockor H7, which suggests that the signal transduction of VIP is probably related with the genic transcription and the PKCdependent phosphorylation of enzyme proteins.

关 键 词：支气管上皮细胞 臭氧 血管活性肠肽 细胞保护 

分 类 号：R562.2[医药卫生—呼吸系统]