生姜提取物6-姜酚对人皮肤成纤维细胞增殖、迁移的影响及其机制研究  

Effect of 6-gingerol on proliferation and migration of human skin fibroblasts and underlying mechanism

作　　者：侯春[1] 王肃生[1] 梁刚[1] 张志华[1] 冀航[1] 王平平 施然 李伯辉 Hou Chun;Wang Susheng;Liang Gang;Zhang Zhihua;Ji Hang;Wang Pingping;Shi Ran;Li Bohui(Department of Plastic and Cosmetic Surgery,the First Affiliated Hospital of Guangzhou Medical University,Guangzhou 510120,China)

机构地区：[1]广州医科大学附属第一医院整形外科,广州510120

出　　处：《中华临床医师杂志（电子版）》2021年第9期691-698,共8页Chinese Journal of Clinicians(Electronic Edition)

基　　金：广东省中医药局科研项目(20201209)。

摘　　要：目的研究不同浓度生姜提取物6-姜酚对人皮肤成纤维细胞增殖、迁移能力的影响,并探讨相关机制。方法采用MTT活力试验检测不同浓度6-姜酚(0、25、50、75、100、150、200 mg/L)对皮肤成纤维细胞活性的影响,设空白对照组(人成纤维细胞)、阳性药物组(重组表皮生长因子20 mg/L)、不同浓度(100、50、25 mg/L)6-姜酚处理组。采用流式细胞术检测细胞增殖情况,Transwell实验检测细胞迁移能力,实时荧光定量及蛋白免疫印迹实验检测细胞血小板源性生长因子(PDGF)、血管内皮生长因子(VEGF)、转化生长因子β_(1)(TGF-β_(1))、基质金属蛋白酶2(MMP2)、基质金属蛋白酶9(MMP9)mRNA和蛋白的表达情况。采用方差分析进行多组样本的差异性比较。结果MTT及流式细胞实验结果显示,与空白对照组相比,在50~200 mg/L的范围内随着6-姜酚浓度的增大,皮肤成纤维细胞活性及增殖能力逐渐增加,差异有统计学意义(P<0.05)。Transwell结果显示,与空白对照组相比,100、50 mg/L 6-姜酚处理组的穿膜细胞数明显增多(P均<0.01),而25 mg/L 6-姜酚处理组的迁移细胞数量与空白对照组比较差异无统计学意义(P>0.05)。与空白对照组相比,100、50 mg/L 6-姜酚处理组的细胞羟脯氨酸分泌表达量明显升高(P均<0.01),而25 mg/L 6-姜酚处理组的细胞羟脯氨酸分泌表达量与空白对照组比较,差异无统计学意义(P>0.05)。实时定量聚合酶链反应结果显示,不同浓度6-姜酚作用于皮肤成纤维细胞24 h后,100、50 mg/L 6-姜酚处理组PDGF、VEGF、MMP2、MMP9 mRNA表达水平较空白对照组显著升高,差异均有统计学意义(P<0.05);100 mg/L 6-姜酚处理组TGF-β_(1) mRNA表达水平较空白对照组显著升高,差异均有统计学意义(P<0.05)。蛋白免疫印迹结果显示,各浓度6-姜酚处理组PDGF、VEGF、MMP9蛋白表达水平均较空白对照组显著升高,差异均有统计学意义(P<0.05);100、50 mg/L 6-姜酚处理组TGF-Objective To investigate the effect of different concentrations of ginger extract 6-gingerol on the proliferation and migration of human skin fibroblasts and explore the possible mechanisms involved.Methods MTT assay was used to detect the effect of different concentrations of 6-gingerol(0,25,50,75,100,150 and 200 mg/L)on the viability of skin fibroblasts.The cells were then divided into a blank control group(untreated human fibroblasts),a positive drug group(recombinant epidermal growth factor 20 mg/L),and 6-gingerol treatment groups(100,50,and 25 mg/L).Flow cytometry was used to detect cell proliferation.Transwell assay was used to detect cell migration ability.Real-time PCR and Western blot were used to detect the mRNA and protein expression of platelet-derived growth factor(PDGF),vascular endothelial growth factor(VEGF),transforming growth factorβ_(1)(TGF-β_(1)),matrix metalloproteinase 2(MMP2)and matrix metalloproteinase 9(MMP9),respectively.Analysis of variance was used to compare the differences among multiple groups of samples.Results MTT assay and flow cytometry showed that compared with the blank control group,as the concentration of 6-gingerol increased in the range of 50-200 mg/L,the activity and proliferation ability of skin fibroblasts increased gradually,with statistically significant differences(P<0.05).Transwell assay showed that compared with the blank control group,the numbers of migrated cells in the 100 and 50 mg/L 6-gingerol treatment groups were significantly increased(P<0.01 for both),while that in the 25 mg/L 6-gingerol treatment group had no statistically significant change(P>0.05).Compared with the blank control group,the secretion of cellular hydroxyproline in the 100 and 50 mg/L 6-gingerol treatment groups was significantly increased(P<0.01 for both),while in the 25 mg/L 6-gingerol treatment group there was no statistically significant difference(P>0.05).Real-time PCR results showed that after treatment of skin fibroblasts with different concentrations of 6-gingerol for 24 h,the exp

关 键 词：6-姜酚 人皮肤成纤维细胞 生长因子 基质金属蛋白酶 

分 类 号：R285[医药卫生—中药学]