Research Of Shenlingbaizhu Powder On Ceramide Metabolic Pathway In Skin Of Mice With Mechanical Barrier Dysfunction

Objective:This study investigated the effect of Shenlingbaizhu Powder that is the representative prescription of fortifying spleen on skin mechanical barrier dysfunction and explored the mechanism.This may provide evidences for applying Jianpi fortifying spleen method to treat skin diseases with barrier dysfunction.Methods:1.The effect of Shenlingbaizhu Powder on skin repair in mice with mechanical barrier dysfunctionICR mice were divided randomly into blank group,model group,and three Shenlingbaizhu Power groups.The three Shenlingbaizhu Powder groups were given Shenlingbaizhu Powder decoction at 10 g/kg,20 g/kg,40 g/kg respectively once a day from the first day,while other groups were administered intragastrically with distilled water once a day for 13 days.From the 9th day,the mouse model of skin mechanical barrier dysfunction was duplicated by tape stripping method twice a day for 5 days.On the day before modeling,1 day after modeling,and 5 days after modeling,TEWL,skin hydration,skin p H,and skin elasticity values were measured using a skin physiology tester.After modeling for 5 days,all the animals were executed to obtain the skin tissue on back and neck.HE staining was performed to observe the morphological changes of the skin tissue,and the thickness of the epidermis was measured.The effect of Shenlingbaizhu Powder on skin repair of mice with mechanical barrier dysfunction was evaluated.2.Mechanism of Shenlingbaizhu Powder for skin repair of mice with mechanical barrier dysfunctionThe animal grouping,modeling and administration were same as those in the first part.After modeling for 5 days,all animals were executed to obtain the skin tissue on back and neck.The protein and m RNA expression of the relevant enzymes and transporters(ABCA12)of the ceramide metabolic pathway were examined.The effect of Shenlingbaizhu Powder on the metabolic pathway of ceramide was investigated to explore the mechanism.The method was as follows: the protein expression of enzymes related to the ceramide biosynthesis pathway(SPT and SMase)and the catabolic pathways(CDase,CK,SMS,GCS)were detected by Western Blot.The protein expression of ABCA12 and enzymes(?-GCase and SMase)related to the ceramide salvage pathway were detected by immunohistochemistry.The ceramide synthetase(SPT and SMase),decomposition enzyme(CDase,CK,SMS,GCS),ABCA12,and enzymes(?-GCase and SMase)related to ceramide salvage pathways were detected by Real-time PCR method.Results:1.The effect of Shenlingbaizhu Powder on skin repair in mice with mechanical barrier dysfunctionThere was no significant difference in TEWL,hydration,p H,and elasticity(R2)of skin among all the groups on the day before modeling.The skin TEWL and p H of the model group increased,and the degree of hydration and elasticity decreased 1 day after modeling.All the Shenlingbaizhu Powder groups can reduce the skin TEWL value,increase the skin hydration degree,and decrease the skin p H of model mice significantly 5 days after modeling.Shenlingbaizhu Powder dosed at 40 g/kg can increase skin elasticity value of mice.All the Shenlingbaizhu Powder groups can improve the status of dermatopathological morphology in various degrees,such as reducing thickness of the stratum corneum,alleviating the acanthosis,and reducing dermal inflammation cells infiltration significantly.The epidermal thickness of mouse skin was reduced in all the Shenlingbaizhu groups.2.The effect of Shenlingbaizhu Powder on ceramide biosynthesis pathway in skin of mice with mechanical barrier dysfunction5 days after modeling,Shenlingbaizhu Powder in all the doses had no significant effect on the protein and m RNA expression of enzymes(SPT and SMase)related to skin ceramide synthesis pathway in mice with mechanical barrier dysfunction.3.Effect of Shenlingbaizhu Powder on ceramide catabolic pathway in mice with skin mechanical barrier dysfunction5 days after modeling,Shenlingbaizhu Powder dosed at 20 g/kg can up-regulate protein and m RNA expression of CDase,CK,and SMS2 in skin of mice with mechanical barrier dysfunction significantly.Shenlingbaizhu Powder dosed at 40 g/kg can increase the skin machinery GCS protein and m RNA expression in skin of mice with barrier dysfunction significantly.Shenlingbaizhu Powder in all the doses had no significant effect on the protein and m RNA expression of SMS1 in skin of mice with mechanical barrier dysfunction.4.The effect of Shenlingbaizhu Powder on ceramide salvage pathway in skin of mice with mechanical barrier dysfunction5 days after modeling,Shenlingbaizhu Powder dosed at 20 g/kg can up-regulate the protein and m RNA expression of ABCA12 and ?-GCase in the skin of mice with mechanical barrier dysfunction significantly.There was no significant effect on protein and m RNA expression of SMase in mouse skin.Conclusions:Shenlingbaizhu Powder can repair the mechanical barrier function of skin,and the mechanism may be related to increase the content of skin ceramides by regulating the related enzymes CDase,CK,SMS2 and ?-GCase and transporter ABCA12 in the ceramide metabolic pathway.