抑制环氧化酶-2-前列腺素E2通路介导硫化氢保护人皮肤角质形成细胞对抗化学性缺氧引起的损伤  

Inhibition of COX-2/PGE2 pathway mediates the protection of hydrogen sulfide against chemical hypoxia- induced injury to human skin keratinocytes

作　　者：杨春涛[1] 董颀[1] 韩艳芳[2] 张辉[3] 郭润民[4] 胡芬[4] 冯鉴强[4] 莫利求[5] 

机构地区：[1]广州医学院生理教研室,510182 [2]中山大学附属孙逸仙纪念医院皮肤科 [3]第三军医大学附属西南医院全军肝胆外科研究所西南肝胆外科医院肝胆外科 [4]中山大学中山医学院生理教研室 [5]中山大学附属第一医院黄埔院区麻醉科

出　　处：《中华生物医学工程杂志》2012年第3期177-181,共5页Chinese Journal of Biomedical Engineering

基　　金：广东省科技计划（2010B080701035）

摘　　要：目的探讨环氧化酶（COX）-2-前列腺素（PG）E2通路在硫化氢（H2S）保护人皮肤角质形成细胞（HaCaT细胞）对抗化学性缺氧引起的损伤中的作用。方法用化学性缺氧模拟剂氯化钴（CoCl2）处理HaCaT细胞，建立缺氧引起皮肤损伤的体外模型。在CoCl2处理前，用不同浓度的H2S的供体硫氢化钠（NaHS）预处理30min，检测H2S对CoCl2引起的细胞损伤的影响；应用细胞计数试剂盒（CCK）-8法检测细胞存活率。酶联免疫法（ELISA）检测培养基中PGE2的水平。Western印迹法检测COX-1和COX-2蛋白的表达。结果500μmol／L CoCl2处理HaCaT细胞24h可以明显降低细胞存活率[（56．3±5．4）％]，促进PGE2的释放[（395．7±65．0）％]以及上调COX-2的表达（均P〈0．01）。CoCl2处理对HaCaT细胞内COX-1的表达无明显影响（P〉0．05）。在200～400μmol／L的浓度范围内，NaHS预处理可明显拮抗CoCl2引起的细胞存活率降低、PGE2的释放增加以及COX-2表达上调。选择性COX-2抑制剂（NS-398）也能抑制CoCl2处理引起的细胞存活率降低。结论H2S可保护人皮肤角质形成细胞对抗化学性缺氧引起的损伤，其机制与抑制COX-2-PGE2通路有关。Objective To investigate the role of COX- 2/PGE2 pathway in protective effects of hydrogen sulfide against chemical hypoxia-induced injury to human skin keratinocytes （HaCaT cells）. Methods The hypoxia-mimicking agent CoCl2 was used to process HaCaT cells and establish an in vitro model of hypoxia-indueed skin injury. Before the exposure to CoCl2, HaCaT cells were pretreated with NariS （a donor of H2S） for 30 rains. The impact of H2S on CoCl2-induced cell injury was then detected. The cell survival rate was determined by cell counting kit（CCK）-8, the expression level of PGE2 in culture medium by enzyme-linked immunosorbent assay （ELISA） and the expression levels of COX-1 and COX-2 protein by Western blot assay. Results The 24 h treatment of HaCaT cells with 500 μmol/L CoCl2 could significantly lower the cell survival rate [ （56.3±5.4）% ], promote the release of PGE2 [ （395.7±65.0）% ] and up-regulate the expression of COX-2 （all P〈0.01）. However, the treatment using CoCl2 did not obviously affect the expressions of COX-1 in HaCaT cells （P〉0.05）. The pretreatment with NariS （ranging from 200 to 400 μmol/L） could significantly inhibit the lowered cell survival, increased release of PGE2 and up-regulated expression of COX-2 caused by CoCl2. Additionally, NS-398, a selective inhibitor of COX-2, could also inhibit CoCl2-induced decrease in cell survival rate. Conclusions H2S can antagonize the chemical hypoxia- induced injury and protect HaCaT cells. Furthermore, this mechanism is associated with the inhibition of COX-2/PGE2 pathway.

关 键 词：硫化氢 角蛋白细胞 环氧化酶-2 前列腺素E2 

分 类 号：R363[医药卫生—病理学]