竹红菌素A通过哺乳动物雷帕霉素靶蛋白依赖途径促进瘢痕疙瘩成纤维细胞自噬的研究  

Hypocrellin A promotess autophagy of keloid fibroblasts through mTOR-dependent pathway

作　　者：陈雷刚[1] 任慧敏[2] 吴远慧[1] 安国芝[1] 景晓蕾 赵同心 CHEN Leigang;REN Huimin;WU Yuanhui;AN Guozhi;JING Xiaolei;ZHAO Tongxin(Department of Dermatology,The First Affliated Hospital of Hebei North University,Zhangjiakou 075000,China;Department of Emergency,The First Affiliated Hospital of Hebei North University,Zhangjiakou 075000,China;Graduate School,Hebei North University,Zhangjiakou 075000,China)

机构地区：[1]河北北方学院附属第一医院皮肤科,河北张家口075000 [2]河北北方学院附属第一医院急诊科,河北张家口075000 [3]河北北方学院研究生学院,河北张家口075000

出　　处：《临床皮肤科杂志》2024年第10期577-581,共5页Journal of Clinical Dermatology

基　　金：河北省中医药管理局科研计划项目(2023105)资助项目。

摘　　要：目的:探究竹红菌素A(HA)通过哺乳动物雷帕霉素靶蛋白(mTOR)依赖途径促进瘢痕疙瘩成纤维细胞(KFs)自噬的作用机制。方法:原代培养KFs并进行传代,将第3代KFs分为对照组、不同剂量HA组(分别予0.125、0.25、0.5、1.0μmol/L HA处理)、si-阴性对照(NC)组、si-NC+HA组(转染NC si RNA+1.0μmol/L HA)及si-磷酸酯酶与张力蛋白同源物(PTEN)+HA组(转染PTEN siRNA+1.0μmol/L HA)。采用CCK8法检测细胞增殖情况;采用酶联免疫吸附试验检测胶原代谢指标Ⅰ型胶原(Col-Ⅰ)、α-平滑肌肌动蛋白(α-SMA)、纤维连接蛋白(FN)的含量;采用western blot检测自噬标志物微管相关蛋白1的轻链3(LC3)-Ⅱ/LC3-Ⅰ、Beclin-1及m TOR途径中PTEN、磷酸化磷脂酰肌醇3-激酶(p-PI3K)、磷酸化蛋白激酶B(p-AKT)、p-mTOR的表达水平。结果:不同剂量HA组KFs的A_(450)值,Col-Ⅰ、α-SMA、FN含量以及p-PI3K、p-AKT、p-mTOR的表达水平均低于对照组;而PTEN、LC3-Ⅱ/LC3-Ⅰ、Beclin-1表达水平均高于对照组(P<0.05),且呈剂量依赖性。si-PTEN+HA组KFs的A_(450)值,Col-Ⅰ、α-SMA、FN含量以及p-PI3K、p-AKT、p-mTOR的表达水平均高于si-NC+HA组(P<0.05);而PTEN、LC3-Ⅱ/LC3-Ⅰ、Beclin-1表达水平均低于si-NC+HA组(P<0.05)。结论:HA通过调控m TOR途径介导的细胞自噬可抑制KFs增殖。Objective:To investigate the role and mechanism of hypocrellin A(HA)in promoting autophagy of keloid fibrob-lasts(KFs)through mammalian rapamycin target protein(mTOR)dependent pathways.Methods:KFs were primarily cultured and passaged.The 3rd generation of KFs were divided into control group,four dose HA groups(0.125,0.25,0.5,1.0μmol/L HA treatment),si-NC group transfected with negative control siRNA,si-NC+HA group transfected with NC siRNA and treat-ed with 1.0μmol/L HA,si-PTEN+HA group transfected with PTEN siRNA and treated with 1.0μmol/L HA.The optical den-sity A45o of cell proliferation was detected by CCK8 assay;the contents of collagen metabolism index type I collagen(Col-I),α-smooth muscle actin(α-SMA)and fibronectin(FN)were detected by ELISA;the expression levels of the autophagy markers light chain 3(LC3)-II/LC3-I,Beclin-1 as well as PTEN,p-PI3K,p-AKT and p-mTOR in mTOR pathways were detected by western blot.Results:The A4so levels,the contents of Col-I,α-SMA,FN,the expression levels of p-PI3K,p-AKT and p-mTOR of dfferent dose HA groups were lower than those in the control group,while the expression levels of PTEN,LC3-II/LC3-I and Beclin-1 were higher than those in the control group(P<0.05),and the level of difference was dose-dependent.The A45o levels,the contents of Col-I,α-SMA,FN,the expression levels of p-PI3K,p-AKT and p-mTOR of si-PTEN+HA group were higher than those in the si-NC+HA group,while the expression levels of PTEN,LC3-II/LC3-I and Beclin-1 were lower than those in the si-NC+HA group(P<0.05).Conclusion:HA inhibits the proliferation of KFs by regulating cell au-tophagy via the mTOR pathway.

关 键 词：瘢痕疙瘩 成纤维细胞 竹红菌素A 哺乳动物雷帕霉素靶蛋白 自噬 

分 类 号：R758.23[医药卫生—皮肤病学与性病学]