长波紫外线照射对人皮肤成纤维细胞表达一氧化氮合成酶和一氧化氮的影响  

The Expression of iNOS and NO in Human Fibroblasts by UVA Irradiation

作　　者：李薇[1] 谢红付[2] 张锡宝[1] 朱慧兰[1] 刘玉梅[1] 罗权[1] 

机构地区：[1]广州市皮肤病防治所,广东广州510095 [2]中南大学湘雅医院皮肤科,湖南长沙410008

出　　处：《中国皮肤性病学杂志》2009年第11期708-710,729,共4页The Chinese Journal of Dermatovenereology

摘　　要：目的通过检测UVA照射后人皮肤成纤维细胞的细胞形态及诱导型一氧化氮合成酶(iNOS)和一氧化氮(NO)表达,初步探讨UVA引起成纤维细胞光损伤的发病机制。方法以1J/cm2,5J/cm2和10J/cm2UVA分别照射人成纤维细胞后24h,用逆转录聚合酶链反应(RT-PCR)和化学细胞免疫方法检测iNOSmRNA和蛋白表达情况,Griess法检测NO的含量,MTT试验检测细胞增殖能力,倒置显微镜和HE染色观察细胞形态变化。结果10J/cm2UVA照射人成纤维细胞后24h出现皱缩最为明显,其细胞增殖能力(MTT)(OD值,0.2472±0.0328)与其他三组比较明显减退(P均<0.05),且iNOSmRNA(1.0568±0.0778)和iNOS蛋白表达强于1J/cm2和5J/cm2(P均<0.05),同时NO的含量(51.11±2.2760)nmol/mL高于1J/cm2的(32.44±3.1620)nmol/mL和5J/cm2的(41.32±2.4460)nmol/mL(P均<0.05)。结论人成纤维细胞iNOS和NO的表达与UVA照射呈剂量依赖。成纤维细胞的光损伤与iNOS和NO产生有关。Objective To investigate the effects of UVA irradiation on expression of inducible nitric oxide synthase （ iNOS） mRNA, iNOS protein and the change of morphology of Human primary fibroblasts. To approach the mechanism on UVA induce fibroblasts photodamage. Methods Human primary fibroblasts were cultured, irradiated cells with UVA, iNOS mRNA and protein on different dose were measured by RT-PCR and cell immunohistochemistry, respectively. Result iNOS mRNA and protein could not be detected on control, iNOS mRNA and protein of fibroblasts were expressed by irradiation of 1,5,10J/cm^2 UVA, cell morph appered to be shrinkage. The result of MTT（ OD Viability 0.2472 ± 0. 0328 ）, iNOS mRNA （ 0.2472 ± 0. 0328 ）, protein and the concentration of NO （51.11 ±2. 2760） increased by irradiation after 24 hours, however, most obviously expressded at 10J/cm^2 UVA irradiation group, and significantly differences respectively compared with other groups （ P 〈 0.05）. Conclusion iNOS mRNA, protein and NO expressions of Human fibroblasts were invariable dependently on dosage after UVA irradiation. The expression of iNOS and NO were concerned with fibroblasts photodamage.

关 键 词：长波紫外线照射 人成纤维细胞 诱导型一氧化氮合成酶 一氧化氮 

分 类 号：R594.8[医药卫生—内科学] R392.13[医药卫生—临床医学]