圣草酚对TGF-β1诱导的人皮肤成纤维细胞增殖、氧化应激反应及TGF-β1/Smad信号通路活化的影响  

Effect of eriodictyol on proliferation,oxidative stress response and TGF⁃β1/Smad signaling pathway activation of human skin fibroblasts induced by TGF⁃β1

作　　者：张泆琳 武鹏 尹建新 方经纬 张钰琴[2] 吕军影[2] ZHANG Yilin;WU Peng;YIN Jianxin;FANG Jingwei;ZHANG Yuqin;LYU Junying(The First School of Clinical Medicine,Guangxi Medical University,Nanning 530021,Guangxi,China;Department of Traditional Chinese Medicine,the First Affiliated Hospital of Guangxi Medical University,Nanning 530021,Guangxi,China)

机构地区：[1]广西医科大学第一临床医学院,广西南宁市530021 [2]广西医科大学第一附属医院中医科,广西南宁市530021

出　　处：《广西医学》2024年第6期873-881,共9页Guangxi Medical Journal

基　　金：广西自然科学基金重点项目(2018GXNSFDA281047);广西中医药重点学科建设项目(GZXK‑Z‑20‑52);广西研究生教育创新计划项目(YCSW2023213)。

摘　　要：目的 探讨圣草酚对转化生长因子β1(TGF-β1)诱导的人皮肤成纤维细胞(HSF)的增殖、氧化应激反应及TGF-β1/Smad信号通路活化的影响。方法 将HSF分为对照组、模型组、圣草酚低剂量组、圣草酚中剂量组、圣草酚高剂量组、SB-431542组。对照组不做任何处理,模型组用5.0 ng/mL TGF-β1干预24 h,圣草酚低剂量组、圣草酚中剂量组、圣草酚高剂量组分别用40μmol/L、80μmol/L、160μmol/L圣草酚和5.0 ng/mL TGF-β1同时干预24 h,SB-431542组用10μmol/L SB-431542和5.0 ng/mL TGF-β1同时干预24 h。检测各组HSF细胞活力、α平滑肌肌动蛋白(α-SMA)蛋白表达水平、活性氧簇(ROS)含量、超氧化物歧化酶(SOD)活性,TGF-β1、α-SMA、Ⅰ型胶原蛋白(ColⅠ)、Ⅲ型胶原蛋白(Co lⅢ)、基质金属蛋白酶2(MMP2)的mRNA表达量及TGF-β1的蛋白表达水平。检测对照组、模型组、圣草酚中剂量组HSF的Smad2、Smad3、磷酸化Smad2 (p-Smad2)、磷酸化Smad3 (p-Smad3)蛋白表达水平。结果 与对照组相比,模型组的HSF细胞活力及α-SMA蛋白表达水平增加,ROS含量增加而SOD活性降低,TGF-β1、α-SMA、ColⅠ、Co lⅢ和MMP2 mRNA表达量上调,TGF-β1、Smad2、Smad3、p-Smad2、p-Smad3的蛋白表达量亦上调(P<0.05)。与模型组相比,圣草酚各剂量组和SB-431542组HSF细胞活力降低、ROS含量降低、SOD活性增强,且α-SMA、ColⅠ、Co lⅢ、MMP2的mRNA表达量和TGF-β1蛋白表达量下调,圣草酚中剂量组、圣草酚高剂量组和SB-431542组HSF细胞中α-SMA蛋白表达水平降低,圣草酚中剂量组及SB-431542组Smad2、Smad3、p-Smad2、p-Smad3蛋白表达量下调(P<0.05)。圣草酚低剂量组、圣草酚中剂量组、圣草酚高剂量组HSF细胞活力、α-SMA蛋白表达水平、ROS含量依次降低,SOD活性依次增强,α-SMA、ColⅠ、Co lⅢmRNA表达量依次降低(P<0.05)。结论 圣草酚可能通过抑制氧化应激反应及TGF-β1/Smad通路的活化,从而剂量依赖性地抑制TGF-β1诱�Objective To explore the effect of eriodictyol on proliferation,oxidative stress response and TGF‑β1/Smad signaling pathway activation of human skin fibroblasts(HSF)induced by transforming growth factorβ1(TGF‑β1).Methods HSF was assigned to control group,model group,eriodictyol low‑dose group,eriodictyol medium‑dose group,eriodictyol high‑dose group,or SB‑431542 group.The control group did not received any treatment,whereas the model group received intervention for 24 hours with 5 ng/mL TGF‑β1,and the eriodictyol low‑,medium,and high‑dose groups received intervention for 24 hours with 40μmol/L,80μmol/L,and 160μmol/L eriodictyol and 5.0 ng/mL TGF-β1 simultaneously,respectively;in addition,the SB-431542 group received intervention for 24 hours with 10μmol/L SB-431542 and 5.0 ng/mL TGF-β1 simultaneously.The HSF cell activity,α-smooth muscle actin(α-SMA)protein expression,reactive oxygen species(ROS)content,superoxide dismutase(SOD)activity,and mRNA expressions of TGF⁃β1,α⁃SMA,collagen typeⅠ(ColⅠ),collagen typeⅢ(ColⅢ),matrix metalloproteinase 2(MMP2),as well as TGF-β1 protein expression were detected in various groups.The protein expressions of Smad2,Smad3,phosphorylated Smad2(p-Smad2),and phosphorylated Smad3(p-Smad3)of HSF were detected in the control group,the model group,and the eriodictyol medium-dose group.Results Compared with the control group,the model group exhibited increased HSF cell activity and elevatedα-SMA protein expression,increased ROS content while decreased SOD activity,up-regulated mRNA expressions of TGF⁃β1,α⁃SMA,ColⅠ,ColⅢ,and MMP2,and up-regulated protein expressions of TGF-β1,Smad2,Smad3,p-Smad2,and p-Smad3(P<0.05).Compared with the model group,the eriodictyol various-dose groups and the SB-431542 group yielded decreased HSF cell activity,ROS content,while increased SOD activity,and down-regulated mRNA expressions ofα⁃SMA,ColⅠ,ColⅢ,and MMP2,as well as down-regulated protein expression of TGF-β1;in addition,eriodictyol medium-and

关 键 词：皮肤纤维化 圣草酚 人皮肤成纤维细胞 转化生长因子β1 氧化应激 转化生长因子β1/Smad信号通路 

分 类 号：R285.5[医药卫生—中药学]