参苓白术散对皮肤机械屏障功能障碍小鼠神经酰胺再利用途径相关酶β-GCase及ASM表达的影响  

Effect of Shenling Baizhusan on Expressions of β-GCase and ASM of Ceramide Salvage Pathway in Mice with Skin Mechanical Barrier Dysfunction

作　　者：刘媛 王葳[1] 王妍[1] 何丹[1] 黄金红 黄丽佳 孙晓菲[1] LIU Yuan;WANG Wei;WANG Yan;HE Dan;HUANG Jin-hong;HUANG Li-jia;SUN Xiao-fei(Yunnan University of Traditional Chinese Medicine,Kunming 650500,China)

机构地区：[1]云南中医学院

出　　处：《中国实验方剂学杂志》2018年第22期112-119,共8页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：2016年云南省科学技术厅-云南中医学院应用基础研究联合专项资金项目[2017FF116(-027)]

摘　　要：目的：探讨参苓白术散对小鼠皮肤机械屏障功能修复作用及对神经酰胺再利用途径相关酶β-葡糖脑苷脂酶（β-glucocerebrosidase,β-GCase）和酸性鞘磷脂酶（acid sphingomyelinase,ASM）表达的影响。方法：50只SPF级ICR小鼠随机分为正常组、模型组、参苓白术散低、中、高（10,20,40 g·kg-1）组。采用胶带剥离法制备皮肤机械屏障功能障碍小鼠模型。参苓白术散低、中、高剂量组于造模前8 d给予相应剂量参苓白术散水煎液灌胃;其余各组灌胃给予蒸馏水,每天1次,连续13 d。皮肤检测仪测量皮肤皮肤经皮水分丢失（TEWL）值、皮肤水合度及皮肤pH;通过苏木素-伊红（HE）染色观察小鼠皮肤病理组织形态学变化;免疫组化法检测β-GCase,ASM的蛋白表达;实时荧光定量聚合酶链式反应（Real-time PCR）分析检测β-GCase,ASM mRNA的表达。结果：造模5 d后,与正常组比较,模型组小鼠皮肤TEWL及pH显著升高,皮肤水合度显著下降（P〈0.01）;β-GCase蛋白及mRNA的表达明显降低（P〈0.05）;与模型组比较,参苓白术散各剂量组均能降低皮肤TEWL值及pH（P〈0.01）,升高皮肤水合度（P〈0.01）,对皮肤病理组织形态学改变有不同程度的改善;同时参苓白术散中剂量组可上调β-GCase蛋白及mRNA的表达（P〈0.05,P〈0.01）。结论：参苓白术散具有修复皮肤机械屏障功能的作用,其机制可能与上调β-GCase的表达从而增加神经酰胺的含量有关。Objective： To investigate the effect of Shenling Baizhusan（SLBZS） in repairing the skin mechanical barrier function in mice and the expression of β-glucocerebrosidase（β-GCase） and acid sphingomyelinase（ASM） related to ceramide salvage pathway. Method： A total of 50 SPF ICR mice were randomly divided into the blank control group,the model group,the low-dose SLBZS group（10 g·kg-1）,the middle-dose SLBZS group（20 g·kg-1） and the high-dose SLBZS group（40 g·kg-1）. The tape-stripping method was used to replicate the mouse model of the dysfunction of the skin mechanical barrier. The low-dose group,the middle-dose group and the high-dose group were given the corresponding doses of SLBZS by gavage 8 d before modeling. The other groups were given distilled water once a day for 13 days. The transepidermal water loss（TEWL）,skin hydration and skin pH were measured. The pathological changes of skin tissue were observed by hematoxylin-eosin（HE） staining. The protein expressions of β-GCase, ASM were detected by immunohistochemistry. β-GCase and ASM mRNA expressions in skin tissues were detected by Real-time PCR.Result： Compared with the blank control group,the level of TEWL and pH were significantly increased in the model group（P〈0. 05）,and the skin hydration was decreased in the model group（P〈0. 01）. The expression ofβ-GCase was decreased in the model group（P〈0. 05）. Compared with the model group,each dose of SLBZS could decrease the levels of TEWL and pH（P〈0. 01）,and increase the skin hydration（P〈0. 01）. The histopathological changes of the skin were improved to varying degrees. The middle-dose SLBZS group could upregulate the expression of β-GCase（P〈0. 05,P〈0. 01）. Conclusion： SLBZS can repair the mechanical barrier function of skin,which may be related to the up-regulation of β-GCase expression and increase of ceramide content.

关 键 词：参苓白术散 皮肤机械屏障功能 β-葡糖脑苷脂酶 酸性鞘磷脂酶 

分 类 号：R2-0[医药卫生—中医学] R22