艾灸“关元”穴对皮肤光老化模型大鼠皮肤组织氧化应激及自噬相关基因表达的影响  

Effects of Moxibustion at"Guanyuan(CV 4)"on Oxidative Stress and Autophagy-Related Gene Expression of Skin Tissue in Photoaging Model Rats

作　　者：惠倩倩 靖媛 欧阳思洁 游世晶[1] 童伯瑛[1] HUI Qianqian;JING Yuan;OUYANG Sijie;YOU Shijing;TONG Boying(School of Acupuncture,Moxibustion and Tuina,Fujian University of Traditional Chinese Medicine,Fuzhou,350122)

机构地区：[1]福建中医药大学针灸推拿学院,福建省福州市350122

出　　处：《中医杂志》2025年第6期621-628,共8页Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金(82004438);福建省自然科学基金(2023J01869)。

摘　　要：目的探讨艾灸关元穴延缓皮肤光老化衰老的潜在作用机制。方法将32只雄性Wistar大鼠随机分为空白组、模型组、维生素E组和艾灸组,每组8只。除空白组外,其余各组大鼠进行紫外线光照背部皮肤建立皮肤光老化模型。造模结束1周后,艾灸组大鼠艾灸“关元”穴,每天1次,每周灸5天;维生素E组大鼠给予维生素E 25 mg/(kg·d)灌胃,每天1次,每周5天;空白组、模型组和艾灸组大鼠给予等量的生理盐水灌胃;各组干预连续7周。7周后取大鼠背部皮肤组织,检测皮肤组织的含水量;采用HE染色观察皮肤组织学形态;ELISA法检测皮肤组织胶原蛋白Ⅰ(CollagenⅠ)、胶原蛋白Ⅲ(CollagenⅢ)含量;检测皮肤组织中丙二醛(MDA)、谷胱甘肽过氧化物酶(GSH-px)、超氧化物歧化酶(SOD)、过氧化氢(H2O2)和过氧化氢酶(CAT)活性;Western Blot法检测皮肤组织中自噬相关蛋白微管相关蛋白1A/1B轻链3(LC3)、多聚泛素结合蛋白(p62)和自噬特异性基因(Beclin-1)蛋白水平;qRT-PCR法检测皮肤组织中LC3、p62、Beclin-1 mRNA表达;透射电镜观察皮肤自噬小体的形成情况。结果HE染色结果显示,空白组大鼠的皮肤表皮结构规整,厚度均匀;模型组大鼠的皮肤表皮厚薄不一;艾灸组大鼠表皮层规则且平整,厚度较为均匀,真皮层排列紧密;维生素E组大鼠皮肤表皮层较模型组变厚。与空白组比较,模型组大鼠皮肤组织含水量及皮肤组织中CollagenⅠ、CollagenⅢ含量均降低,皮肤组织SOD、CAT和GSH-Px活性均降低,H2O2及MDA活性升高,p62蛋白水平及其mRNA表达升高,LC3和Beclin-1蛋白水平及其mRNA表达降低(P<0.05或P<0.01)。与模型组比较,艾灸组上述各指标均有所改善(P<0.05或P<0.01),维生素E组皮肤组织中CollagenⅠ和CollagenⅢ含量差异无统计学意义(P>0.05)。透射电镜结果显示,与空白组比较,模型组皮肤细胞整体萎缩,线粒体出现大量空泡化,细胞结构降解;艾灸组大鼠皮肤Objective To explore the potential mechanism of moxibustion at Guanyuan(CV 4)in delaying skin photoaging.Methods Thirty-two male Wistar rats were randomly divided into four groups,namely blank group,model group,vitamin E group,and moxibustion group,with 8 rats in each group.Except for the blank group,dorsal skin of rats were exposed to ultraviolet(UV)radiation to establish a skin photoaging model.One week after modeling,the moxibustion group received moxibustion at"Guanyuan(CV 4)"once a day,five days per week;the vitamin E group received vitamin E(25 mg/kg·d)once a day by gavage,five days per week;the blank group,model group,and moxibustion group received an equivalent volume of normal saline via gavage;the intervention lasted for 7 weeks.After 7 weeks,dorsal skin tissues were collected to analyze the following indicators,such as skin tissue moisture content,histomorphological changes using hematoxylin-eosin(HE)staining,CollagenⅠand collagenⅢcontent using ELISA.Malondialdehyde(MDA),glutathione peroxidase(GSH-Px),superoxide dismutase(SOD),hydrogen peroxide(H2O2),and catalase(CAT)activity in skin tissue were dectected.Western Blot was used to determin autophagy-related proteins,including microtubule-associated protein 1A/1B-light chain 3(LC3),polyubiquitin-binding protein(p62),and autophagy-specific gene(Beclin-1);LC3,p62,and Beclin-1 mRNA expression was detected via qRT-PCR,and autophagosome formation was observed using transmission electron microscopy(TEM).Results HE staining showed that the epidermal structure in the blank group was orderly and evenly thick,while the model group exhibited uneven epidermal thickness.In the moxibustion group,the epidermis was well-structured,smooth,and uniform,with densely arranged dermal layers;the epidermis in the vitamin E group was thicker than that in the model group.Compared with the blank group,the model group exhibited decreased skin moisture content and reduced level of CollagenⅠand collagenⅢ,reduced SOD,CAT,and GSH-Px activity in skin tissue,increased H2O2 and M

关 键 词：光老化 皮肤老化 艾灸 关元穴 自噬 氧化应激 胶原蛋白 

分 类 号：R24[医药卫生—中医临床基础]