补益营卫方对衰老表皮ULK1、BECN1、LC3Ⅰ、LC3Ⅱ、ATG3、ATG5、ATG7、ATG9表达的影响  

Effect of Recipe of Supplementing Ying-wei on the expressions of ULK1,BECN1,LC3Ⅰ,LC3Ⅱ,ATG3,ATG5,ATG7 and ATG9 in senescent epidermis

作　　者：李婷 袁浩棓 冯仰磊 周小平[1] LI Ting;YUAN Hao-Bei;FENG Yang-Lei(Traditional Chinese Medicine School of Ningxia Medical University,Yinchuan 750004,Ningxia,China)

机构地区：[1]宁夏医科大学中医学院,宁夏银川750004

出　　处：《中国老年学杂志》2024年第16期3937-3941,共5页Chinese Journal of Gerontology

基　　金：国家自然科学基金项目(82160910)。

摘　　要：目的观察补益营卫方对衰老小鼠皮肤表皮自噬相关基因(ATG)1在哺乳动物中的同源基因(ULK1)、ATG6在哺乳动物中同源基因(BECN1)、微管相关蛋白轻链(LC)3Ⅰ、LC3Ⅱ、ATG3、ATG5、ATG7、ATG9表达的影响,探讨补益营卫方延缓皮肤衰老的机制。方法选取10只SPF级3月龄SAMR1雄性小鼠作为年轻组,取40只SPF级8月龄SAMP8雄性小鼠随机分为4组,每组10只,分别设为模型组和(补益营卫方)高、中、低剂量组。年轻组和模型组灌服等剂量生理盐水,高、中、低剂量组每天分别按10.0、5.0、2.5 g/kg的剂量灌胃给药,每周禁食称重1次,连续6 w后脱臼处死小鼠,褪毛并取背部皮肤组织,分离表皮细胞裂解后取上清液,采用实时荧光定量聚合酶链反应(PCR)法检测表皮细胞ULK1、BECN1、LC3Ⅰ、LC3Ⅱ、ATG3、ATG5、ATG7、ATG9 mRNA表达量;免疫组织化学法评价表皮ULK1、BECN1、LC3Ⅰ、LC3Ⅱ、ATG3、ATG5、ATG7、ATG9蛋白表达。结果与年轻组相比,模型组皮肤表皮ULK1、ATG7、ATG9、LC3ⅡmRNA及蛋白表达显著升高(P<0.05,P<0.01);与模型组相比,高剂量组ULK1、ATG7、ATG9、LC3ⅡmRNA及蛋白表达、中剂量组LC3ⅡmRNA、ATG7蛋白表达、低剂量组ATG7蛋白表达差异有统计学意义(P<0.05,P<0.01)。结论补益营卫方通过抑制ULK1、ATG7、ATG9、LC3ⅡmRNA及蛋白表达,改善衰老表皮细胞的自噬活性,发挥延缓皮肤衰老的作用。Objective To observe the effect of Recipe of Supplementing Ying-wei on the expressions of homologs of autophagy related genes(ATG)1 in mammals(ULK1),ATG6 homologous genes in mammals(BECN1),microtubule-associated protein light chain(LC)3Ⅰ,LC3Ⅱ,ATG3,ATG5,ATG7 and ATG9 in skin epidermal of aging mice,and to explore the mechanism of Recipe of Supplementing Ying-wei in delaying skin aging.Methods Ten SPF 3-month-old SAMR1 male mice were selected as the young group,and 40 SPF 8-month-old SAMP8 male mice were randomly divided into 4 groups,with 10 mice in each group,mice were divided into model group and(Recipe of Supplementing Ying-wei)high,midum and low dose groups,respectively.Mice in the young group and model group were given equal doses of normal saline,and those in high,medium and low dose groups were given 10.0,5.0 and 2.5 g/kg Recipe of Supplementing Ying-wei,respectively,fasting and weighing once a week for six consecutive weeks.Then the mice were dislocated and sacrificed,hair was removed and the epidermal cells were lysed to obtain the supernatant.The mRNA expressions of ULK1,BECN1,LC3Ⅰ,LC3Ⅱ,ATG3,ATG5,ATG7 and ATG9 in epidermal cells were detected by real-time fluorescence quan-titative polymerase chain reaction(PCR)method.The expressions of epidermal ULK1,BECN1,LC3Ⅰ,LC3Ⅱ,ATG3,ATG5,ATG7 and ATG9 protein were evaluated by immunohistochemistry.Results Compared with the young group,the mRNA and potein expressions of ULK1,ATG7,ATG9 and LC3Ⅱin the skin epidermal of the model group were significantly increased(P<0.05,P<0.01);compared with the model group,there were statistical significance in the mRNA and potein expressions of ULK1,ATG7,ATG9 and LC3Ⅱof high dose group,LC3ⅡmRNA and ATG7 potein expressions of medium dose group,ATG7 potein expression of low dose group(P<0.05,P<0.01).Conclusions Recipe of Supplementing Ying-wei could inhibit the expressions of ULK1,ATG7,ATG9,LC3ⅡmRNA and protein,improve the autophagy activity of aging epidermal cells,and play a role in delaying skin aging.

关 键 词：补益营卫方 皮肤衰老 表皮细胞 自噬 

分 类 号：R285.5[医药卫生—中药学]