摘要
目的研究大成散对体外培养人皮肤成纤维细胞(HFF-1)增殖及鞘氨醇激酶/1-磷酸鞘氨醇(SPHK/S1P)通路的影响,并初步探讨其作用机制。方法制备大成散药液,采用CCK-8法检测不同浓度大成散药液对HFF-1细胞增殖活性的影响,确定最佳干预时间与浓度进行后续实验,Western Blot法检测大成散药液对HFF-1细胞转化生子因子-β1(TGF-β1)、SPHK1、SPHK2、S1P蛋白表达的影响,qRT-PCR法检测大成散药液对HFF-1细胞TGF-β1、SPHK1、SPHK2 mRNA表达的影响。结果5×10^(-2)g/L的大成散干预48h可促进HFF-1细胞的增殖,提高细胞活力;与对照组比较,大成散可以增加HFF-1细胞中TGF-β1、SPHK1、SPHK2、S1P蛋白表达水平(P<0.05),提高HFF-1细胞中TGF-β1、SPHK1、SPHK2 mRNA表达水平(P<0.05)。结论大成散可通过促进成纤维细胞增殖,上调TGF-β1表达,激活SPHK1/S1P通路,促进血管新生,发挥抗炎效应,或通过激活SPHK2,催化产生S1P,启动缺氧预处理机制,从而保护皮肤成纤维细胞,提高成纤维细胞的增殖能力,进而加速创面愈合。
Objective To study the effect of Dacheng Powder on the proliferation of human foreskin fibroblasts-1(HFF-1)and sphingosine kinase/Sphingosine 1-phosphate(SPHK/S1 P)pathway of human skin fibroblasts cultured in vitro,and to preliminarily study its mechanism of action.Methods Dacheng Powder liquid was prepared.The effects of different concentrations of Dacheng Powder liquid on the proliferation activity of HFF-1 cells were detected by Cell Counting Kit-8(CCK-8),and the optimal intervention conditions were determined for subsequent experiments.Western Blot was used to detect the effects of Dacheng Powder liquid on expressions of transforming growth factor beta1(TGF-β1),sphingosine kinase(SPHK1),SPHK2,sphingosine 1-phosphate(S1 P)protein.Quantitative Real-time polymerase chain reaction(qRT-PCR)method was used to detect Dacheng Powder on mRNA expressions of TGF-β1,SPHK1,and SPHK2.Results Dacheng Powder at 5×10^(-2)g/L promoted the proliferation of HFF-1 cells and improved cell viability.Compared with the control group,Dacheng Powder increased the protein expressions of TGF-β1,SPHK1,SPHK2,and S1P in HFF-1 cells(P<0.05),and elevated mRNA expressions of TGF-β1,SPHK1,SPHK2 in HFF-1 cells(P<0.05).Conclusion Dacheng Powder exerted anti-inflammatory effects possibly through promoting the proliferation of fibroblasts,up-regulating the expression of TGF-β1,activating the SPHK1/S1P pathway,promoting angiogenesis,or protecting skin fibroblasts and improving the proliferation of fibroblasts,thereby accelerating wound healing possibly through activating SPHK2,catalyzing the production of S1P,and initiating the mechanism of hypoxia preconditioning.
作者
陈慧玲
江莉
官美龄
陈艺玲
翁慧兰
黄宁
CHEN Hui-ling;JIANG Li;GUAN Mei-ling;CHEN Yi-ling;WENG Hui-lan;HUANG Ning(Department of Medical Cosmetology,Xiamen Hospital,Bejing University of Chinese Medicine,Fujian 361006;Department of Dermatology,Second People's Hospital Affiliated to Fujian University of Traditional Chinese Medicine,Fujian Provincial Key Laboratory of Integrated Traditional Chinese and Western Medicine Dermatology,Fujian 350001)
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2024年第6期733-737,共5页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家中医药管理局第二届国医大师传承工作室建设项目(No.国中医药办人教发[2015]24号)
福建省自然科学基金项目(No.2021 J01882)
福建省科技计划项目(引导性项目)(No.2020 Y0044)
福建省中医学术流派传承工作室建设项目(No.闽卫中医函[2019]129号]。
关键词
创面愈合
大成散
成纤维细胞
鞘氨醇激酶/1-磷酸鞘氨醇信号通路
中药复方
wound healing
Dacheng Powder
fibroblast
sphingosine kinase/sphingosine 1-phosphate signaling pathway
Chinese herbal compound
