Spleen And Skin Mechanical Barrier Function Of The Relationship Between Some Experimental Studies

PurposeThe splenic asthenia mouse models and pruritus mouse model induced by impairment of cutaneous permeability barrier function are copied in company with, The scratching behavior are observed,the content of ceramides is determined and the thickness of epidermis is determined.Observe the effect of traditional Chinese drug Huanglianjiedu Decoction,Senling Baishu Powder to the mouse models,Because of traditional Chinese medicine clinical regularity,we'd like to study the relationship between traditional Chinese drug for invigorating the spleen and atopic dermatitis, splenic asthenia and cutaneous permeability barrier function by the experimental study,and provide new research ideas and methods for the study about the hypostasis of splenic asthenia.Methods1.Epidermal lipids extraction of mouse and evaluating the method for determining the content of epidermal ceramidesEpidermal lipids of mouse are extracted by routine method,separated by thin-layer chromatography(TLC),shot by BIO-RAD GelDoc system.Epidermal ceramides is determined by Quantity One.And then evaluate this methodology.2.The study of pruritus mouse model induced by impairment of cutaneous permeability barrier function.To select the pruritus model for using,the rostral skin of mouse is defatted by organic solvent to make the pruritus model,the scratching behavior are observed and the content of ceramides is determined after making the pruritus model for 0day, 3days,5 days and 7 days.3.The effect of Huanglianjiedu Decoction to splenic asthenia models and pruritus mouse model induced by impairment of cutaneous permeability barrier function.The two splenic asthenia mouse models are induced by Rhubarb,Reserpine.And then pruritus model is copied on the two splenic asthenia mouse models. Huanglianjiedu Decoction is administrated.The scratching frequency is recorded, the content of ceramides is determined and the thickness of epidermis is determined.4.The effect of Huanglianjiedu Decoction to the mouse model of cutaneous permeability barrier function and the interference effect of ShenLing BaiZhu Decoction.The splenic asthenia mouse models are induced by Huanglianjiedu Decoction, And then pruritus model is copied on the splenic asthenia mouse models. ShenLingBaiZhu Decoction is administrated as treatment.The scratching frequency is recorded,the content of ceramides is determined and the thickness of epidermis is determined.Results1.Epidermal lipids extraction of mouse and evaluating the method for determining the content of epidermal ceramidesThe ceramides content and density score have good linear relationship when the ceramides content is between 0.416μg and 2.08μg,R=0.9953;The precision of the same plate is good,RSD=1.09%;The precision of different plates is good,RSD= 2.10%;The stability of control article solution and sample solution is good within 2h. The reproducibility is good,RSD=3.03%;The average recovery is 99.58%, RSD=1.46%.The RSD of simple determination is 3.10%.2.The study of pruritus mouse model induced by impairment of cutaneous permeability barrier functionThe scratching behavior is most significant after making the pruritus model for 5 days,P＜0.001,vs control group.The average contents of ceramides are lower after making the pruritus model for 5 days,P＜0.001,vs model for 0day.3.The effect of Hnanglianjiedu Decoction to splenic asthenia models and pruritus mouse model induced by impairment of cutaneous permeability barrier function. Scratching behavior Compare to the control group,The average scratching frequency of pruritus model mice increase in evidence(P＜0.01).The average scratching frequency of two splenic asthenia pruritus model mice is less than the scratching frequency of pruritus model mice.Reserpine group P＜0.05.The average scratching frequency of medication administration team is less than the average scratching frequency of two splenic asthenia pruritus model mice.Ceramide content①Splenic asthenia induced by Rhubarb,average:control group＞pruritus group＞Rhubarb group＞medication administration team.P＜0.001, pruritus group vs control group.P＜0.05,Rhubarb group vs pruritus group, medication administration team vs Rhubarb.②Splenic asthenia induced by Reserpine average:controlgroup＞pruritus group＞Reserpine group＞medication administration team.P＜0.001,pruritus group vs control group,Reserpine group vs pruritus.Ceramide content of medication administration team comparing with it of pruritus group is not improve.Thickness of epidermis After copying the pruritus mouse model,the epidermis thickens.P＜0.001,vs control group.Splenic asthenia induced by Rhubarb and Reserpine can aggravate the thickening of epidermal.P＜0.05,vs control group.The average epidermal thickness of Rhubarb model mouse is less than the one of pruritus model mouse.The average epidermal thickness of medication administration team mouse is less than the one of corresponding splenic asthenia model mouse, comparing with two group is not statistically significant.4.The effect of Huanglianjiedu Decoction to the mouse model of cutaneous permeability barrier function and the interference effect of ShenLing BaiZhu Decoction.Scratching behavior Compare to the control group,The average scratching frequency of pruritus model mice increase in evidence(P＜0.01).Splenic asthenia induced by Huanglianjiedu Decoction is copied,The average scratching frequency of the splenic asthenia pruritus model mice is less than the scratching frequency of pruritus model mice(P＜0.05).The average scratching frequency of medication administration team is less than the average scratching frequency of the splenic asthenia pruritus model(P＜0.05)Ceramide content average:control group＞pruritus group＞medication administration team＞Huanglianjiedu Decoction group.P＜0.001,pruritus group vs control group.P=0.052,Huanglianjiedu Decoction group vs pruritus group,P＜0.05,medication administration team vs Huanglianjiedu Decoction group.Thickness of epidermis After copying the pruritus mouse model,the epidermis thickens.P＜0.001,vs control group.Splenic asthenia induced by Huanglianjiedu Decoction can aggravate the thickening of epidermal.P＜0.05,vs pruritus group. ShenLingBaiZhu decoction can inhibit the thickening of epidermis significantly.The average epidermal thickness of medication administration team mouse is less than the one of Huanglianjiedu Decoction model mouse,P＜0.001.ConclusionsThe experimental use of epidermal ceramides content,scratching behavior, Thickness of epidermis as a testing target,The pruritus mouse model induced by impairment of cutaneous permeability barrier function is copied,compared with the control group,The scratching behavior is most significant,the content of epidermal ceramides is reducible,the epidermis thickens.According to the results,The epidermal ceramides content will decrease when the cutaneous permeability barrier function is destroyed.Splenic asthenia will aggravate the decrease of ceramides and the cutaneous permeability barrier disorders.The epidermal ceramides content of the three splenic asthenia models mice induced by Rhubarb,Reserpine and Huanglianjiedu Decoction is decreased.The three splenic asthenia models can make the poor cutaneous permeability barrier function poorer.ShenLingBaiZhu Decoction can improve it.