豨莶消白饮调控沉默信息调节因子2同源蛋白1/过氧化物酶体增殖物激活受体γ共激活剂-1α通路对白癜风模型小鼠治疗作用及机制研究

Therapeutic effect and mechanism of Xixian Xiaobai Beverage on vitiligo model mice through regulating silent information regulator 1/peroxisome proliferator-activated receptor γ coactivator-1α pathway

目的探讨豨莶消白饮对白癜风模型小鼠皮肤毛皮脱色情况、炎症程度、沉默信息调节因子2同源蛋白1(SIRT1)/过氧化物酶体增殖物激活受体γ共激活剂-1α(PGC-1α)通路蛋白表达的影响。方法将40只SPF级6周龄C57BL/6健康雄性小鼠随机分为对照组、模型组、阳性药物组和豨莶消白饮组,模型组、阳性药物组和豨莶消白饮组以5%的过氧化氢(H2O2)溶液涂擦50 d诱导小鼠白癜风模型,豨莶消白饮组于造模21 d以147 mg/(kg·d)的豨莶消白饮灌胃干预30 d,阳性药物组灌胃464 mg/(kg·d)白癜风胶囊,观察小鼠皮毛脱色情况,Masson-Fontana染色观察并计算表皮黑色素表达面积百分率和黑色素分布评分,统计含黑色素毛囊率,采用酶联免疫吸附试验(ELISA)检测血清酪氨酸酶(TYR)、丙二醛(MDA)、超氧化物歧化酶(SOD)和8-羟基脱氧鸟苷(8-OHDG)表达水平,采用蛋白质免疫印迹(Western blot)检测皮肤组织SIRT1、PGC-1α和锰超氧化物歧化酶(MnSOD)蛋白表达水平。结果H2O2溶液诱导的模型组小鼠皮肤表皮内黑色素表达面积百分率和表皮黑色素分布评分、黑色素毛囊率、血清TYR和SOD的含量、皮肤组织MnSOD、SIRT1、PGC-1α蛋白表达低于对照组,MDA、8-OHdG表达高于对照组,差异有统计学意义(P<0.05)。豨莶消白饮组黑色素表达面积百分率和表皮黑色素分布评分、黑色素毛囊率、血清TYR和SOD的含量、皮肤组织MnSOD、SIRT1、PGC-1α蛋白表达高于对照组,MDA、8-OHdG表达低于模型组,差异有统计学意义(P<0.05)。豨莶消白饮组和模型组皮肤组织MnSOD蛋白表达比较,差异无统计学意义(P>0.05)。结论H2O2溶液诱导的白癜风小鼠氧化应激增强可能与SIRT1/PGC-1α信号转导通路的改变有关;豨莶消白饮能通过调节SIRT1/PGC-1α信号通路,增强皮肤组织抗氧化能力,防止白癜风的发生发展。

Objective To explore the effect of Xixian Xiaobai Beverage on skin and fur decolorization,inflammation,and expression of silent information regulator 1(SIRT1)/peroxisome proliferator-activated receptor γ coactivator-1α(PGC-1α)pathway proteins on vitiligo model mice and the underlying mechanism.Methods A total of 40 SPF only 6 weeks of health male C57BL/6 mice were randomly divided into control group,model group,positive drug group and Xixian Xiaobai Beverage group.Model group,positive drug group and Xixian Xiaobai Beverage group were established by embrocating 5% hydrogen peroxide(H2O2)for 50 consecutive days.The Xixian Xiaobai Beverage group were given Xixian Xiaobai Beverage 147 mg/(kg·d)by gavage on the 21st day of modeling.The positive drug group were given Baidianfeng Capsules 464 mg/(kg·d)by gavage.The skin and hair decolorization were observed and recorded,Masson-Fontana staining was used to observe and calculate the percentage of melanin expression area and melanin distribution score of the epidermis.The rate of melanin-containing hair follicles was statistically calculated.Enzyme-linked immunosorbent assay(ELISA)was used to detect the expression levels of serum tyrosinase(TYR),malondialdehyde(MDA),superoxide dismutase(SOD)and 8-hydroxydeoxyguanosine(8-OHDG).Western blot was used to detect the protein expression levels of SIRT1,PGC-1αand manganese SOD(MnSOD)in skin tissues.Results The percentage of epidermal melanin expression area,epidermal melanin distribution score,melanin hair follicle rate,contents of TYR and SOD in serum,expressions of MnSOD,SIRT1 and PGC-1α in model group induced by H2O2 solution were lower than those in control group,while the expressions of MDA and 8-OHdG were higher than those in control group,the differences were statistically significant(P<0.05).The percentage of melanin expression area,epidermal melanin distribution score,melanin hair follicle rate,contents of TYR and SOD in serum,expressions of MnSOD,SIRT1 and PGC-1α in skin tissue in Xixian Xiaobai Beverage group were higher than those in control group,while the expressions of MDA and 8-OHdG were lower than those in model group,with statistical significance(P<0.05).There was no significant difference in MnSOD protein expression between Xixian Xiaobai Beverage group and model group(P>0.05).Conclusion Enhancement of oxidative stress induced by H2O2 solution in vitiligo mice may be related to the change of SIRT1/PGC-1α pathway.Xixian Xiaobai Beverage can enhance the antioxidant capacity of skin tissues and prevent the occurrence and development of vitiligo by regulating the SIRT1/PGC-1αsignaling pathway.