蜕皮甾酮对皮肤光老化和黑色素生成的影响  

Effects of ecdysterone on skin photoaging and melanogenesis

作　　者：龙剑文 张晨 李杨玺宇 LONG Jianwen;ZHANG Chen;LI Yangxiyu(The First Clinical School of Hubei University of Chinese Medicine,Wuhan 430061,Hubei,China;Clinical College of Traditional Chinese Medicine,Hubei University of Chinese Medicine,Wuhan 430061,Hubei,China)

机构地区：[1]湖北中医药大学第一临床学院,湖北武汉430061 [2]湖北中医药大学中医临床学院,湖北武汉430061

出　　处：《现代中西医结合杂志》2023年第16期2197-2202,2215,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：第五批全国中医临床优秀人才研修项目(国中医药人教函[2022]1号)。

摘　　要：目的观察蜕皮甾酮对皮肤光老化和黑色素生成的影响。方法①实验分为正常对照组、1.0μmol/L蜕皮甾酮组、1.5μmol/L蜕皮甾酮组、2.0μmol/L蜕皮甾酮组,各组分别处理角质形成细胞(HaCaT细胞)、黑素细胞(B16F10细胞)和人真皮成纤维细胞(HDF细胞)后,MTT法检测细胞活力。②实验分为正常对照组、UVB组、1.0μmol/L蜕皮甾酮组、1.5μmol/L蜕皮甾酮组、2.0μmol/L蜕皮甾酮组,Annexin V-FITC/PI双标法检测HaCaT细胞凋亡情况,Hoechst 33258染色法检测凋亡细胞的形态,Western blot法检测HaCaT细胞中基质金属蛋白酶-1(MMP-1)、基质金属蛋白酶-9(MMP-9)、环氧化酶-2(COX-2)、沉默调节蛋白1(Sirt-1)表达情况。③实验分为正常对照组、1.0μmol/L蜕皮甾酮组、1.5μmol/L蜕皮甾酮组、2.0μmol/L蜕皮甾酮组,Western blot法检测HaCaT细胞中透明质酸合成酶2(HAS-2)、转谷氨酰胺酶1(TGM1)和HDF细胞中Ⅰ型胶原蛋白Iα1肽链(COL1A1)表达情况。④实验分为正常对照组、黑色细胞刺激素组、1.0μmol/L蜕皮甾酮组、1.5μmol/L蜕皮甾酮组、2.0μmol/L蜕皮甾酮组、熊果苷组,采用分光光度仪检测B16F10细胞黑色素生成及分泌情况。⑤实验分为正常对照组、左旋多巴组、1.0μmol/L蜕皮甾酮组、1.5μmol/L蜕皮甾酮组、2.0μmol/L蜕皮甾酮组,采用分光光度仪检测各组蘑菇酪氨酸酶活性。结果不同浓度的蜕皮甾酮对HaCaT细胞、B16F10细胞、HDF细胞活力均无明显影响。蜕皮甾酮各组HaCaT细胞凋亡率及HaCaT细胞中MMP-1、MMP-9、COX-2蛋白相对表达量均明显低于UVB组(P均<0.05),HaCaT细胞中Sirt-1蛋白相对表达量均明显高于UVB组(P均<0.05)。蜕皮甾酮各组HaCaT细胞中HAS-2、TGM1蛋白相对表达量和HDF细胞中COL1A1蛋白相对表达量均明显高于正常对照组(P均<0.05)。蜕皮甾酮各组和熊果苷组B16F10细胞黑色素生成和分泌吸光度值均明显低于黑色细胞刺激素组(P均<0.05)。蜕皮甾酮各组蘑菇Objective It is to observe the effects of ecdysterone on skin photoaging and melanogenesis.Methods①The experiment was divided into normal control group,1.0μmol/L ecdysterone group,1.5μmol/L ecdysterone group,2.0μmol/L ecdysterone group,after the keratinocytes(HaCaT cells),melanocytes(B16F10 cells),and human dermal fibroblasts(HDF cells)were treated in each group,the cell viability in each group was detected by MTT method.②The experiment was divided into normal control group,UVB group,1.0μmol/L ecdysterone group,1.5μmol/L ecdysterone group,and 2.0μmol/L ecdysterone group,the apoptosis of HaCaT cells was detected by Annexin V-FITC/PI double-labeling assay,the morphology of apoptotic cells was detected by Hoechst 33258 staining assay,and the expressions of matrix metalloproteinase-1(MMP-1),matrix metalloproteinase-9(MMP-9),cyclo-oxygenase-2(COX-2),and silencing regulator protein 1(Sirt-1)in HaCaT cells were detected by Western blot method.③The experiments was divided into normal control group,1.0μmol/L ecdysterone group,1.5μmol/L ecdysterone group and 2.0μmol/L ecdysterone group,the expressions of hyaluronan synthase 2(HAS-2),transglutaminase 1(TGM1)in HaCaT cells,and peptide chain of collagen type I Iα1 in HDF cells were detected by Western blot method.④The experiment was divided into normal control group,melanocyte stimulating hormone group,1.0μmol/L ecdysterone group,1.5μmol/L ecdysterone group,2.0μmol/L ecdysterone group and arbutin group,and the production and secretion of melanin in B16F10 cells were detected by spectrophotometer.⑤The experiment was divided into normal control group,levodopa group,1.0μmol/L ecdysterone group,1.5μmol/L ecdysterone group and 2.0μmol/L ecdysterone group,the mushroom tyrosinase activity of each group was detected by spectrophotometer.Results Different concentrations of ecdysterone had no significant effect on the viability of HaCaT cells,B16F10 cells and HDF cells.The apoptosis rate of HaCaT cells and the relative expressions of MMP-1,MMP-9 and COX-2 prot

关 键 词：蜕皮甾酮 皮肤衰老 黑色素生成 

分 类 号：R-33[医药卫生]