基于PKC/CREB/MITF通路探讨白藜芦醇对紫外线辐照后人皮黑素细胞增殖及黑素合成的影响  

Effects of resveratrol on proliferation and melanin synthesis of human skin melanocytes after ultraviolet irradiationbased on PKC/CREB/MITF pathway

作　　者：李婷婷 周燕 尹明丹 刘秀琴 冉春涛 LI Ting-ting;ZHOU Yan;YIN Ming-dan;LIU Xiu-qin;RAN Chun-tao(Department of Dermatology,Banan Hospital Affiliated to Chongqing Medical University,Chongqing,401320,China;Department of Dermatology,People's Hospital of Kaizhou District,Chongqing,405400,China)

机构地区：[1]重庆医科大学附属巴南医院皮肤科,重庆401320 [2]重庆市开州区人民医院皮肤科,重庆405400

出　　处：《职业与健康》2022年第22期3054-3060,共7页Occupation and Health

基　　金：重庆市开州区科委开科发号(20182)。

摘　　要：目的基于蛋白激酶C(the cAMP signaling pathway activates protein kinase C,PKC)/cAMP反应元件结合蛋白(cAMP response element binding protein,CREB)/小眼相关转录因子(ommatidium-associated transcription factor,MITF)通路探讨白藜芦醇对紫外线辐照后人皮黑素细胞增殖及黑素合成的影响。方法设人表皮黑素细胞组、紫外线A段(uv A segment,UVA)暴露组、白藜芦醇低、高剂量组,除人表皮黑素细胞组外,其余各组暴露于320~400 nm的日光紫外线辐照,UVA辐照结束后白藜芦醇低和高剂量组加入不同浓度溶解于胎牛血清(fetal bovine serum,FBS)的白藜芦醇(300、600μg/mL),UVA暴露组加入等体积的FBS。以上各组每孔设6个平行样,培养72 h。用Cell Counting Kit-8及甲基紫染色测定法测量细胞增殖及单克隆形成数目,流式细胞仪测定细胞凋亡水平,酶标仪测定细胞黑素和酪氨酸酶水平,实时荧光逆转录法(real-time fluorescent reverse transcription,RT-PCR)及蛋白印迹法测定细胞PKC、CREB和MITF水平。结果与人表皮黑素细胞组比较,UVA暴露组、白藜芦醇低、高剂量组OD值、存活率、单细胞克隆形成数目、黑素合成能力、酪氨酸酶水平、PKC、CREB、MITF mRNA蛋白表达水平升高,凋亡率降低,差异均有统计学意义(均P<0.05);与UVA暴露组比较,白藜芦醇低、高剂量组OD值、存活率、单细胞克隆形成数目、黑素合成能力、酪氨酸酶水平、PKC、CREB、MITF mRNA蛋白表达水平降低,凋亡率升高,差异均有统计学意义(均P<0.05);与白藜芦醇低剂量组比较,白藜芦醇高剂量组OD值、存活率、单细胞克隆形成数目、黑素合成能力、酪氨酸酶水平、PKC、CREB、MITF mRNA蛋白表达水平降低,凋亡率升高,差异均有统计学意义(均P<0.05)。结论白藜芦醇对紫外线辐照后人皮黑素细胞增殖及黑素合成具有明显抑制作用,其机制可能与白藜芦醇阻断PKC/CREB/MITF信号通路的激活有关。Objective To investigate the effect of resveratrol on the proliferation and melanin synthesis of human skin melanocytes after ultraviolet irradiation based on the cAMP signaling pathway activates protein kinase C(PKC)/cAMP response element binding protein(CREB)/ommatidium-associated transcription factor(MITF)pathway.Methods Human epidermal melanocyte group,uv A segment(UVA)exposure group,low and high dose resveratrol groups were established.Except for the human epidermal melanocyte group,the rest of the groups were exposed to solar ultraviolet radiation of 320-400 nm.After UVA irradiation,different concentrations of resveratrol(300,600μg/mL)dissolved in FBS were added to the low-dose and high-dose groups of resveratrol,and the same volume of FBS was added to the UVA exposure group.Six parallel samples were set for each well of the above groups and cultured for 72 hours.The Cell Counting Kit-8 and methyl violet staining assay were used to measure cell proliferation and the number of monoclonal formation.The flow cytometry was used to determine the level of cell apoptosis.The microplate reader was used to determine the levels of melanin and tyrosinase.The real-time fluorescent reverse transcription(RT-PCR)and Western blotting were used to determine the levels of PKC,CREB and MITF in cells.Results Compared with human epidermal melanocyte group,the OD value,surv ival rate,number of single cell clones,melanin synthesis ability,tyrosinase level,and expression levels of PKC,CREB and MITF mRNA in UVA exposure group,low-dose and high-dose resveratrolgroups increased,while the survival rate decreased,and the differences were statistically significant(all P<0.05).Compared with UVA exposure group,the OD value,survival rate,number of single cell clones,melanin synthesis ability,tyrosinase level,and expression levels of PKC,CREB and MITF mRNA in low-dose and high-dose resveratrol groups decreased,and the differences were statistically significant(all P<0.05).Compared with low-dose resveratrol group,the OD value,survival rate,

关 键 词：基于蛋白激酶C CAMP反应元件结合蛋白 小眼相关转录因子 白藜芦醇 紫外线 人皮黑素细胞 

分 类 号：R758.1[医药卫生—皮肤病学与性病学]