Oxidative stress-induced posttranslational modification of TRPV1 expressed in esophageal epithelial cells
Oxidative stress-induced posttranslational modification of TRPV1 expressed in esophageal epithelial cells
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DOI：
10.1152/ajpgi.00436.2009
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发表时间：
2011-08-01
影响因子：
4.5
通讯作者：
Yoshikawa, Toshikazu
中科院分区：
文献类型：
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作者：
Kishimoto, Etsuko;Naito, Yuji;Yoshikawa, Toshikazu
Kishimoto E，Naito Y，Handa O，Okada H，Mizushima K，Hirai Y，Nakabe N，Uchiyama K，石川T，Takagi T，Yagi N，Kokura S，Yoshida N，Yoshikawa T.氧化应激诱导食管上皮细胞表达TRPV 1的翻译后修饰美国生理学杂志胃肠和肝脏生理学301：G230-G238，2011年。首次发表于2011年6月2日; doi：10.1152/ajpgi.00436.2009。人食管上皮持续暴露于物理刺激或胃酸，有时会引起粘膜炎症。瞬时受体电位香草酸1（TRPV 1）是一种可被辣椒素、热和质子激活的伤害性、钙选择性离子通道。据报道，食管粘膜中表达的TRPV 1的激活参与胃食管反流病（GERD）或非糜烂性GERD症状。在这项研究中，我们研究了TRPV 1在人食管上皮细胞系Het 1A中的表达和功能，特别关注氧化应激的作用。用ELISA法检测Het 1A细胞在辣椒素或酸（含/不含4-羟基-2-壬烯醛（HNE））刺激下分泌的白细胞介素-8（IL-8）。辣椒素刺激后，使用氧化还原敏感的荧光探针测定细胞内活性氧（ROS）的产生，并分别使用生物素化半胱氨酸和抗HNE抗体通过Western印迹法测定ROS和HNE修饰的蛋白质。用合成的HNE处理后，通过免疫沉淀进一步研究TRPV 1蛋白的HNE修饰。辣椒素和辣椒酸诱导Het 1A细胞产生IL-8，TRPV 1拮抗剂可降低这种产生。辣椒素还显著增加了Het 1A细胞内ROS和ROS或HNE修饰蛋白的产生。此外，辣椒素刺激的Het 1A细胞中IL-8的产生通过合成HNE处理而增强。免疫沉淀研究表明，TRPV 1被修改的HNE在合成HNE刺激的Het 1A细胞。我们的结论是，TRPV 1的功能在食管上皮细胞的趋化因子的生产，这一功能可能是通过TRPV 1的翻译后修饰的ROS调节。
Kishimoto E, Naito Y, Handa O, Okada H, Mizushima K, Hirai Y, Nakabe N, Uchiyama K, Ishikawa T, Takagi T, Yagi N, Kokura S, Yoshida N, Yoshikawa T. Oxidative stress-induced posttranslational modification of TRPV1 expressed in esophageal epithelial cells. Am J Physiol Gastrointest Liver Physiol 301: G230-G238, 2011. First published June 2, 2011; doi:10.1152/ajpgi.00436.2009.-Human esophageal epithelium is continuously exposed to physical stimuli or to gastric acid that sometimes causes inflammation of the mucosa. Transient receptor potential vanilloid 1 (TRPV1) is a nociceptive, Ca2+-selective ion channel activated by capsaicin, heat, and protons. It has been reported that activation of TRPV1 expressed in esophageal mucosa is involved in gastroesophageal reflux disease (GERD) or in nonerosive GERD symptoms. In this study, we examined the expression and function of TRPV1 in the human esophageal epithelial cell line Het1A, focusing in particular on the role of oxidative stress. Interleukin-8 (IL-8) secreted by Het1A cells upon stimulation by capsaicin or acid with/without 4-hydroxy-2-nonenal (HNE) was measured by ELISA. Following capsaicin stimulation, the intracellular production of reactive oxygen species (ROS) was determined using a redox-sensitive fluorogenic probe, and ROS- and HNE-modified proteins were determined by Western blotting using biotinylated cysteine and anti-HNE antibody, respectively. HNE modification of TRPV1 proteins was further investigated by immunoprecipitation after treatment with synthetic HNE. Capsaicin and acid induced IL-8 production in Het1A cells, and this production was diminished by antagonists of TRPV1. Capsaicin also significantly increased the production of intracellular ROS and ROS- or HNE-modified proteins in Het1A cells. Moreover, IL-8 production in capsaicin-stimulated Het1A cells was enhanced by synthetic HNE treatment. Immunoprecipitation studies revealed that TRPV1 was modified by HNE in synthetic HNE-stimulated Het1A cells. We concluded that TRPV1 functions in chemokine production in esophageal epithelial cells, and this function may be regulated by ROS via posttranslational modification of TRPV1.