没食子酸调控TLR4/NF-κB通路对UVB诱导人永生化角质形成细胞光损伤的保护作用  

The Protective Effect of Gallic Acid on UVB Induced Photodamage in Human Immortalized Keratinocytes by Regulating the TLR4/NF-κB Pathway

作　　者：陈丽美 都日娜 张敏 崔博 高耀星[4] CHEN Limei;DU Rina;ZHANG Min;CUI Bo;GAO Yaoxing(Inner Mongolia Medical University,Hohhot O10l10,China;Department of Dermatology,Inner Mongolia Autonomous Region China Mongolia Medical Research Institute,Hohhot 010010,China;Department of Dermatology,Mengzhong Hospital,Hohhot 010000,China;Department of Pain,Inner Mongolia Medical University Affiliated Hospital,Hohhot 010050,China)

机构地区：[1]内蒙古医科大学,呼和浩特010110 [2]内蒙古自治区中蒙医药研究院皮肤科,呼和浩特010010 [3]内蒙古呼和浩特市蒙中医院皮肤科,呼和浩特010000 [4]内蒙古医科大学附属医院疼痛科,呼和浩特010050

出　　处：《中国细胞生物学学报》2025年第2期195-202,共8页Chinese Journal of Cell Biology

摘　　要：该文探究没食子酸调控Toll样受体4(Toll-like receptor 4,TLR4)/核因子κB(nuclear factor kappa-B,NF-κB)通路对紫外线B(UVB)诱导人永生化角质形成细胞光损伤的保护作用。将HaCaT细胞分为空白组(未经任何处理)、UVB组(仅暴露于UVB)、没食子酸不同浓度(5、10和20μg/mL)组、没食子酸(20μg/mL)+脂多糖(10μg/mL)组,除空白组外,其余细胞均于相应药物处理后暴露于UVB。MTT法检测细胞活性;试剂盒检测超氧化物歧化酶(superoxide dismutase,SOD)活性和丙二醛(malondialdehyde,MDA)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、白细胞介素-6(interleukin-6,IL-6)水平;TUNEL染色检测细胞凋亡;DCFH-DA探针和显色底物分别检测活性氧(reactive oxygen species,ROS)、半胱天冬蛋白酶(cysteinyl aspartate specific proteinase,caspase)-3和caspase-9活性;蛋白质免疫印迹法检测TLR4、NF-κB、诱导型一氧化氮合酶(inducible nitric oxide synthase,iNOS)和前列腺素内过氧化物合酶2(cyclooxygenase-2,COX-2)蛋白表达情况。将小鼠分为空白组、UVB组、50 mg/kg没食子酸组和没食子酸+脂多糖组,使用UVB诱导UVB组、50 mg/kg没食子酸组和没食子酸+脂多糖组小鼠皮肤光损伤,用50 mg/kg没食子酸治疗。与空白组比较,UVB组HaCaT细胞存活率和SOD活性降低(P<0.05),ROS荧光强度、MDA、炎性因子(TNF-α和IL-6)水平、细胞凋亡指标(TUNEL阳性细胞率、caspase-3和caspase-9活性)、TLR4、p-NF-κB、iNOS和COX-2蛋白水平升高(P<0.05);与UVB组比较,没食子酸不同浓度组HaCaT细胞存活率和SOD活性升高(P<0.05),ROS荧光强度、MDA、炎性因子水平、细胞凋亡指标、TLR4、p-NF-κB、iNOS和COX-2蛋白水平降低(P<0.05);与20μg/mL没食子酸组比较,没食子酸+脂多糖组HaCaT细胞存活率和SOD活性降低(P<0.05),ROS荧光强度、MDA、炎性因子水平、细胞凋亡指标、TLR4、p-NF-κB、iNOS和COX-2蛋白水平升高(P<0.05)。50 mg/kg没食子酸组小鼠皮肤状态优于UVB组;�This paper was to investigate the protective effect of gallic acid on UVB(ultraviolet B)induced photodamage in human immortalized keratinocytes by regulating the TLR4(Toll-like receptor 4)/NF-κB(nuclear factor kappa B)pathway.HaCaT cells were grouped into blank group(untreated),UVB group(exposed only to UVB),gallic acid in different concentrations(5,10 and 20μg/mL)groups,and gallic acid(20μg/mL)+lipopolysaccharide(10μg/mL)groups.Except for the blank group,all other cells were exposed to UVB after corresponding drug treatment.MTT method was used to detect cell viability.Kit method was used to detect the activity of SOD(superoxide dismutase)and the levels of MDA(malondialdehyde),TNF-α(tumor necrosis factor-α),and IL-6(interleukin-6),respectively.TUNEL staining method was used to detect cell apoptosis.DCFH-DA probe and chromogenic substrate were used to detect ROS(reactive oxygen species),caspase(cysteinyl aspartate specific proteinase)-3,and caspase-9 activities,respectively.Immunoblotting method was used to detect the protein expression of TLR4,NF-κB,iNOS(inducible nitric oxide synthase),and COX-2(cyclooxygenase-2).The mice were divided into blank,UVB,50 mg/kg gallic acid and gallic acid+lipopolysaccharide groups,and UVB induced photodamage in the skin of mice in the UVB,50 mg/kg gallic acid and gallic acid+lipopolysaccharide groups,which were treated with 50 mg/kg gallic acid.Compared with the blank group,the HaCaT cell survival rate and SOD activity in the UVB group were lower(P<0.05),and the ROS fluorescence intensity,MDA,inflammatory factor(TNF-αand IL-6)levels,apoptosis index(TUNEL positive cell rate,caspase-3 and caspase-9 activities),TLR4,p-NF-κB,iNOS and COX-2 protein levels were higher(P<0.05).Compared with the UVB group,the HaCaT cell survival rate and SOD activity were higher in the gallic acid different concentration groups(P<0.05),and the ROS fluorescence intensity,MDA,inflammatory factor levels,apoptosis index,TLR4,p-NF-κB,iNOS and COX-2 protein levels were lower(P<0.05).Compared with the 2

关 键 词：没食子酸 Toll样受体4 核因子ΚB 角质形成细胞 光损伤 

分 类 号：R758.1[医药卫生—皮肤病学与性病学]