摘要
目的观察中波紫外线(UVB)辐射后永生化角质形成细胞株(HaCaT细胞)中光产物环丁烷嘧啶二聚体(CPDs)的产生和清除情况,观察川芎嗪对UVB辐射后细胞光产物的影响并探讨其保护作用机制。方法采用30mJ/cm2UVB照射培养的HaCaT细胞,加入川芎嗪干预处理,采用免疫组织化学法检测CPDs,以RT-PCR法和Western blotting法检测各受试组中p53和增殖细胞核抗原(PCNA)的mRNA和蛋白的表达水平。结果UVB辐射后HaCaT细胞中CPDs生成,0.5h达高峰,辐射后4h内清除速率较快,4h后清除速率较慢直至辐射24h。川芎嗪处理UVB辐射的细胞中CPDs少于单纯照射组(P<0.05)。川芎嗪可下调p53(34.9%)和PCNA(30.9%)的mRNA表达,还可下调p53(23.1%)和PCNA(24.9%)的蛋白表达。结论HaCaT细胞对UVB照射所致光产物CPDs的清除存在快速期及慢速期;川芎嗪可降低光产物CPDs水平。川芎嗪的光保护作用可能与其下调修复相关调控分子p53和PCNA基因及蛋白表达有关。
Objective To observe the production and removal of photo-product, cyclobutane pyrimidine dimers （CPDs）, in HaCaT cells induced by UVB irradiation and intervention effect of ligustrazine, and to investigate its photo-protective mechanisms on HaCaT cells damaged from UVB irradiation. Methods HaCaT cells were cultured or irradiated with UVB irradiation of 30 mJ/cm^2 and then treated with ligustrazine. The production and removal of CPDs were examined by immunohistochemical method. The mRNA and protein expressions of p53 and proliferating cell nuclear antigen （PCNA） were detected by RT-PCR and Western blotting assay, respectively. Results CPDs appeared in HaCaT cells, reached the peak at 0. 5 h and removed rapidly during the first 4 h, and then removed slowly until 24 h after UVB irradiation. The quantity of CPDs in HaCaT cells under UVB irradiation in ligustrazine-treated group was less than that in merely irradiated group （P〈0.05）. The mRNA expression of p53 （34. 9%） and PCNA （30. 9%） was down regulated, and so were the protein expressions of p53 （23.1%） and PCNA （24. 9%）, by ligustrazine, respectively. Conclusion There seems to be two phases in the removal of CPDs： a rapid phase and a slower phase after UVB irradiation. Ligustrazine could decrease the level of CPDs. Down-regulation of the mRNA and protein expressions of DNA damage and repair related proteins p53 and PCNA may be a part of photo-protective mechanisms of ligustrazine.
出处
《中草药》
CAS
CSCD
北大核心
2007年第6期878-882,共5页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金资助项目(30371294)
