橙皮素对小鼠皮肤急性光损伤的保护作用  

Protective Effect of Hesperetin on Skin Photodamage Induced by Ultraviolet Radiation B in Mice

作　　者：卢倩[1] 卢志禹 程雨欣 夏青月 骆丹[1] Lu Qian;Lu Zhiyu;Cheng Yuxin;Xia Qingyue;Luo Dan(The First Affiliated Hospital of Nanjing Medical University,Nanjing 210000,China)

机构地区：[1]南京医科大学第一附属医院,江苏南京210000

出　　处：《中国中西医结合皮肤性病学杂志》2022年第3期199-205,共7页Chinese Journal of Dermatovenereology of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金项目(编号:81972961)。

摘　　要：目的探讨橙皮素对中波紫外线(UVB)导致的小鼠皮肤光损伤的抑制作用及可能机制。方法不同剂量UVB照射小鼠背部皮肤,构建UVB致小鼠皮肤急性光损伤的动物模型。将40只小鼠随机分为4组,即空白组、UVB组、基质+UVB组、橙皮素+UVB组,每组各10只。除空白组外,其余每组小鼠每天以相同剂量照射UVB,连续3 d,末次照射后24 h取照射区皮肤组织。肉眼观察皮肤外观变化,光学显微镜观察组织病理学改变,免疫荧光检测皮肤组织活性氧(ROS)水平,水溶性四唑盐(WST-1)法测定总超氧化物歧化酶(SOD)活力,硫代巴比妥酸(TBA)法测定丙二醛(MDA)蛋白表达量。实时荧光聚合酶链反应(RT-PCR)检测核因子E2相关因子2(Nrf2)、Kelch样环氧氯丙烷相关蛋白-1(Keap1)、血红素氧合酶-1(HO-1)的信使核糖核酸(mRNA)表达量,免疫组化检测Nrf2蛋白表达水平,蛋白质印迹法(WB)检测Keap1、HO-1蛋白表达水平。结果选择辐照度3.6 mJ/(cm^(2)·s)照射UVB 200 s,连续3 d建立小鼠皮肤急性光损伤模型。预先外用橙皮素乳膏后,受照射小鼠皮肤组织ROS活力、MDA水平低于UVB组及基质组,SOD水平、Nrf2、Keap1及HO-1的mRNA水平和蛋白表达均高于其余组,差异有统计学意义(P<0.05)。结论外用橙皮素能够减轻UVB照射导致的小鼠皮肤氧化应激反应,从而减轻急性光损伤,其机制可能与Nrf2信号通路有关。Objective In order to investigate the inhibitory effects and possible mechanisms of hesperetin on skin photodamage induced by ultraviolet radiation b(UVB)in mice.Methods Different doses of UVB irradiated mouse back skin,and an animal model of acute mouse skin light injury caused by UVB was constructed.Forty mice were randomly divided into four groups,including blank group,UVB group,matrix+UVB group and hesperetin+UVB group with 10 mice in each group.Except for the blank group,mice in each group were irradiated with UVB at the same dose for three days,and skin tissue from the irradiated area was taken 24 h after the last irradiation.Skin appearance changes were observed visually,histopathological changes were observed by light microscopy.The levels of reactive oxygen species(ROS)were measured by immunofluorescence.Superoxide dismutase(SOD)vitality was measured by 2-(4-iodophenyl)-3-(4-nitriphenyl)-5(2,4-disulfophenyl)-2H-tetrazolium(WST-1).The protein expression level of malondialdehyde(MDA)was determined by thiobarbituric acid(TBA).The mRNA expression level of nuclear factor erythroid-2 related factor 2(Nrf2),Kelch-like epichlorohydrin-associated protein 1(Keap1),and Heme oxygenase-1(HO-1)was determined by reverse transcriptionpolymerase chain reaction(RT-PCR).The Nrf2 protein expression levels were determined by immunohistochemistry.The protein expression levels of Keap1 and HO-1 were determined by Western blotting.Results The skin of the mouse was exposed to UVB with 200 s for three consecutive days.The irradiance of UVB was 3.6 mJ/(cm^(2)·s).After external application of hesperetin cream,ROS viability and MDA levels were lower than UVB and matrix+UVB groups,while SOD levels,mRNA levels and protein expression of Nrf2,Keap1 and HO-1 and were higher than the remaining groups,the differences were statistically significant(P<0.05).Conclusion Hesperetin is able to reduce the skin oxidative stress response in mice caused by UVB irradiation,thus reducing the acute light injury.The mechanism may be related to the Nrf2

关 键 词：橙皮素 光损伤 氧化应激 核因子E2相关因子2 活性氧 

分 类 号：R751[医药卫生—皮肤病学与性病学]