黑色素代谢途径在香叶木素改善紫外诱导的ARPE-19细胞损伤中的保护作用  

The protective role of melanin metabolism pathway in improvement of UV-induced ARPE-19 cell damage by diosmetin

作　　者：马绍壮 冀欣业 张晏源 张波 MA Shao-zhuang;JI Xin-ye;ZHANG Yan-yuan;ZHANG Bo(School of Pharmacy,Shihezi University,Key Laboratory of Xinjiang Phytomedicine Resources and Utilization,Ministry of Education,Shihezi,Xinjiang 832000,China;School of Pharmacy,Chengdu University,Sichuan Institute of Antibiotic Industry,Chengdu 610106,China)

机构地区：[1]石河子大学药学院,新疆植物药资源利用教育部重点实验室,新疆石河子832000 [2]成都大学药学院,四川抗菌素工业研究所,四川成都610106

出　　处：《中国药理学通报》2024年第11期2061-2068,共8页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No U1603122);四川省天府峨眉创新领军人才项目(No 1811)。

摘　　要：目的探讨香叶木素对年龄相关性黄斑变性(age-related macular degeneration,AMD)保护作用及机制。方法基于GEO数据库分析AMD与正常病例样本视网膜色素上皮(retinal pigment epithelial,RPE)细胞差异表达基因。建立302 nm紫外线B(ultraviolet B,UVB)照射人视网膜上皮细胞(human retinal pigment epithelial cells,ARPE-19)细胞损伤模型,评估0.6~40μmol·L^(-1)的香叶木素、10~160μmol·L^(-1)的左旋多巴和1~100μg·L^(-1)的黑色素的药效,并探讨其抗AMD相关通路。检测黑色素含量、酪氨酸酶活性及相关蛋白表达水平。结果经GSE91087筛选发现AMD病例样本组RPE细胞中存在显著差异表达基因,如MCHR1、TYR、TYRP1等,确定黑色素合成及酪氨酸酶代谢通路为AMD的关键通路;100μmol·L^(-1)的黑色素和60μmol·L^(-1)的左旋多巴明显提高紫外照射ARPE-19细胞下的细胞活力,并降低细胞坏死/凋亡的比例;10μmol·L^(-1)的香叶木素提高细胞活力;逆转UVB降低的ARPE-19细胞黑色素含量及酪氨酸酶活性;并提高TYR、TRP-1、TRP-2的蛋白表达,证实了香叶木素在促进黑色素合成方面的作用。结论香叶木素通过酪氨酸代谢途径诱导黑色素的合成,从而改善紫外诱导的ARPE-19细胞损伤。s:Aim To explore the protective effect of diosmetin on age-related macular degeneration(AMD)and its mechanism.Methods Based on the GEO database,the differentially expressed genes between AMD and normal samples of retinal pigment epithelial(RPE)cells were analyzed.The cell damage model of human retinal pigment epithelial cells(ARPE-19)irradiated was established with 302 nm ultraviolet B(UVB).Then the pharmacological effects of 0.6-40μmol·L^(-1) of diosmetin,10-160μmol·L^(-1) of L-DOPA,and 1-100μmol·L^(-1) of melanin were assessed,and their anti-AMD-related pathways were explored.The melanin content,tyrosinase activity,and related protein expression levels were assayed.Results Screened by GSE91087,significant differentially expressed genes such as MCHR1,TYR,and TYRP1 were found in the RPE cells of the AMD case sample group,which determined that melanin synthesis and tyrosinase metabolism pathways were the key pathways in AMD;100μmol·L^(-1) of melanin and 60μmol·L^(-1) of L-DOPA significantly improved cell viability under UVB irradiation of ARPE-19 cells and decreased the necrosis/apoptosis ratio;10μmol·L^(-1) of diosmetin increased cell viability,reversed the reduced melanin content and tyrosinase activity of ARPE-19 cells in the UVB group,and increased the protein expression of TYR,TRP-1,and TRP-2,which confirmed the role of diosmetin in the promotion of melanin synthesis.Conclusion Diometin induces melanin synthesis via tyrosine metabolism pathway and thereby improves UV-induced ARPE-19 cell damage.

关 键 词：年龄相关性黄斑变性 紫外光损伤 香叶木素 黑色素 酪氨酸酶 生物信息学分析 

分 类 号：R285[医药卫生—中药学]