苯乙基间苯二酚对UVB诱导的人皮肤黑素细胞氧化模型损伤的保护作用及其机制  

Protection and Mechanism of Phenylethyl Resorcinol (PR) on UVB-induced Oxidated Damage in PIG1 Cell

作　　者：刘园园[1] 李欣[1] 靳佳慧 王昊[1] 鲍炎 李宁 郭毅群 王艳[1] LIU Yuanyuan;LI Xin;JIN Jiahui;WANG Hao;BAO Yan;LI Ning;Guo Yiqun;WANG Yan(School of Chinese Materia Mediea,Tianjin University of Traditional Chinese Medicine,Tianjin 300193,China;Tianjin Stomatological Hospital,Tianjin 300041,China;Beijing Center For Health Food and Cosmetics Control,Beijing 100200,China;Shang Mei Cosmetics Technology Co,Ltd,Tianjin 300385,China)

机构地区：[1]天津中医药大学,天津300193 [2]天津市口腔医院,天津300041 [3]北京市保健食品化妆品检验中心,北京102200 [4]天津尚美化妆品有限公司,天津300385

出　　处：《中国皮肤性病学杂志》2018年第10期1107-1112,共6页The Chinese Journal of Dermatovenereology

基　　金：国家自然科学基金项目(81602773)

摘　　要：目的探讨苯乙基间苯二酚(PR)对紫外(UVB)诱导的人皮肤黑素细胞(PIG1)氧化模型损伤的保护作用及其机制。方法体外培养PIG1细胞,用UVB处理,建立PIG1UVB氧化损伤模型。用含有不同浓度PR的完全培养基溶液处理PIG1细胞24h;将正常培养的PIG1细胞随机分为正常组、模型组和PR低、中、高剂量组(20、60、100μmol/L),模型组给予UVB照射,给药组紫外照射后给药处理48h。用MTT法检测细胞存活率;用氢氧化钠裂解法检测黑素的变化;用ELISA法检测人优黑素(eumelanin)的变化;用RT-PCR和Western-blot检测与黑素生成相关的基因及蛋白,与酪氨酸酶相关的酪氨酸酶(TYR)、酪氨酸酶相关蛋白1(TRP1)、酪氨酸酶相关蛋白(TRP2)、小眼畸形相关转录因子(MITF),以及氧化相关的蛋白核因子E2相关转录因子2(Nrf2)、NADP(H)醌氧化还原酶1(NQO1)、血红素氧合酶-1(HO-1)的mRNA和蛋白表达情况。结果当PR浓度≤100μmol/L时,细胞存活率不受影响。正常组与模型组比较,模型组黑素与优黑素显著升高;给药组与模型组比较,给药组能够显著抑制黑素和优黑素的生成。RT-PCR和Western blot结果表明,PR能够显著下调Nrf2、TRP1、MITF和NQO1的表达(P <0. 05),对TYR、TRP2和HO-1表达有影响,但与模型组比较差异无统计学意义,相关基因的表达与蛋白表达相一致。结论 PR能保护PIG1细胞对抗UVB诱导的氧化损伤,具有一定的剂量关系,其作用机制可能与抗氧化和抑制酪氨酸酶,降低黑素生成有关。Objective To investigate the protective effect and mechanism of Phenylethyl resor-cinol（PR） on UVB-induced oxidated damage in PIG1 cell. Methods Models of oxidative stress damage were established with the treatment of UVB in vitro. PIG1 cell was treated with different concentrations of PR for 24 h; PIG1 cell was divided into normal group,model group and three PR groups of low,middle and high concentration respectively（20,60,100μmol/L）. Model group and PR group pretreated with UVB,and PR groups were treated with PR for 48 h. The cell viability was detected by MTT assay; melanin and eumelanin were detected by NaOH splitting and ELISA method respectively. Gene and protein expresssion of Nuclear factor-E2-related factor（Nrf2）,Tyrosinase（TYR）,Tyrosinase-related protein1（TRP1）,Tyrosinase-related protein2（TRP2）, Microphthalmia-associaated transcription factor（MITF）,NADP（H） quinone oxidoreductase1（NQO1） and Heme oxygenase-1（HO-1） in PIG1 cells were detected by real time fluorescence quantitative PCR（RT-PCR） and western blot. Results The cell survival rate was not affected when concentration of PR was less than 100μmol/L. Compared with normal group,melanin and eumelanin increased significantly in model group,while they decreased in PR groups compared to model group. In addtion,PR can significantly down-regulate the expression level of Nrf2,TRP1,MITF,NQO1（P〉0. 05） and has no effect on the expression level of TYR,TRP2 and HO-1. Conclusion PR can protect the PIG1 cell against UVB-induced oxidative damage in a certain range. The mechanism may be related to antioxidant,inhibiting tyrosinase and reducing melanogenesis.

关 键 词：苯乙基间苯二酚 PIG1细胞 黑素 优黑素 

分 类 号：R751[医药卫生—皮肤病学与性病学]