摘要
研究沙枣叶(Elaeagnus angustifolia L.)的化学成分和抑制黑色素生成活性。采用硅胶、Sephadex LH-20凝胶、半制备高效液相色谱和薄层色谱等技术对沙枣叶乙醇提取物进行了分离纯化,结合波谱数据和文献报道鉴定化合物的结构,并对化合物体外蘑菇酪氨酸酶(tyrosinase,TYR)抑制活性和细胞内抑制黑色素合成活性进行测定。通过RT-qPCR和Western blot检测化合物对小鼠黑色素瘤细胞B16F10内TYR、TYR相关蛋白1(TYR-related protein 1,TRP-l)、TRP-2和小眼畸形相关转录因子(microphthalmia-associated transcription factor,MITF)mRNA及蛋白表达的影响。采用ELISA法测定化合物处理的B16F10细胞内环磷酸腺苷(cyclic adenosine monophosphate,cAMP)浓度;RT-qPCR、Western blot检测黑色素合成信号通路中蛋白激酶B(protein kinase B,AKT)、磷酸化AKT(p-AKT)、β-连环蛋白(β-catenin)、磷酸化β-catenin(p-β-catenin)的mRNA和蛋白表达水平。从沙枣叶中分离鉴定3个化合物,分别为naphthisoxazol A(1),isorhamnetin-3-O-sophorabioside(2)和isorhamnetin 3-O-β-D-sophoroside-7-O-α-L-rhamnoside(3)。结果发现3个化合物对B16F10细胞内TYR活性具有一定的抑制作用,且降低TYR、TRP-1、TRP-2和MITF基因的转录与蛋白表达。与对照组相比,3个化合物能显著抑制细胞内cAMP产生,降低p-AKT和细胞核中β-catenin含量,最终下调MITF和TYR家族蛋白(TYR、TRP-1和TRP-2)表达,导致细胞内黑色素的合成减少。
This study aims to investigate the chemical constituents and anti-melanogenic activity of Elaeagnus angustifolia L.leaves.The compounds were separated and purified from alcohol extract of E.angustifolia leaves by a series of chromatographic techniques,including silica gel column chromatography,Sephadex LH-20 column chromatography,semi-preparative high performance liquid chromatography and thin-layer chromatography.The structures of the compounds were identified through spectral data and literature reports,their in vitro mushroom tyrosinase(TYR)inhibitory activity and intracellular melanin synthesis inhibitory activity were detected.The regulation effects of the compounds on the mRNA and protein expression of TYR,TYR-related protein 1(TRP-l),TRP-2 and microphthalmia-associated transcription factor(MITF)in mouse melanoma cells B16F10 were detected by RT-qPCR and Western blot.The concentration of cyclic adenosine monophosphate(cAMP)in B16F10 cells treated with the compound was determined by ELISA.RT-qPCR and Western blot were applied to evaluate the mRNA and protein expression levels of melanin synthesis signaling pathways proteins including protein kinase B(AKT),phosphorylated AKT(p-AKT),β-catenin and phosphorylatedβ-catenin(p-β-catenin)in compound-treated B16F10 cells.Three compounds were isolated and identified from the leaves of E.angustifolia,namely naphthisoxazol A(1),isorhamnetin-3-O-sophoroside(2),and isorhamnetin3-O-β-D-sophoroside-7-O-α-L-rhamnoside(3).The results showed that all three compounds could inhibit B16F10 intracellular TYR activity and decrease the transcription and protein expression of TYR,TRP-1,TRP-2,and MITF genes in B16F10 cells.Compared with the control group,the three compounds significantly inhibited intracellular cAMP production and subsequently reduced the protein expression of p-AKT and nuclearβ-catenin,thereby downregulated the expression of MITF and TYR family proteins(TYR,TRP-1 and TRP-2)leading to reduced intracellular melanin synthesis.
作者
徐国燕
付建红
胡泳华
王俊
买迪娜·阿布都外力
崔凤真
阿依卡买尔·艾克拜尔
马畅达
XU Guo-yan;FU Jian-hong;HU Yong-hua;WANG Jun;ABUDUWAILI Maidina;CUI Feng-zhen;AKBAR Aykamar;MA Chang-da(College of Life Science,Xinjiang Normal University,Urumqi 830017,China;School of Biological Science and Technology,Minnan Normal University,Zhangzhou 363000,China)
出处
《天然产物研究与开发》
北大核心
2026年第1期25-36,共12页
Natural Product Research and Development
基金
国家自然科学基金(32260226)
新疆维吾尔自治区“天山创新团队计划”(2022D14004)
新疆维吾尔自治区自然科学基金(2021D01A122)。
