Study On The Pharmacological Activity Of Famous Prescriptions‘The Huanglian Ointment’ In Treating Eczema

Objective:In order to improve the stability and repeatability of the process,digital research on the extraction process of the Huanglian Ointment was carried out on the basis of the ancient recipe process;exploring the target of the Huanglian Ointment in the treatment of eczema based on the method of network pharmacology;through the establishment of a pharmacological model of guinea pig eczema to prove the effectiveness of the Huanglian Ointment and to carry out research on the mechanism of the Huanglian Ointment in the treatment of eczema.Methods:(1)On the basis of methodological investigations,single-factor experiments of medicinal material particle size,frying time,frying temperature,and soaking time were carried out to determine the four-factor level of the orthogonal test,and optimize the process according to the orthogonal test table.(2)On the basis of network pharmacology research methods,the collection of chemical components and the screening of active ingredients,the screening of disease genes,the prediction and analysis of the docking of the target of Huanglian plaster medicinal material and the target of eczema are carried out.(3)The pharmacological model of guinea pig eczema was established,and the skin pathological changes after the model were observed.At the same time,enzyme-linked immunosorbent assay was used to detect the changes of inflammatory factors in guinea pig serum.Immunohistochemical method was used to investigate the changes of C-Jun and FLG protein expression in the skin of guinea pigs.(4)Establish an in vitro eczema cell model,use the MTT method to detect the survival rate of Hacat cells,determine the administration concentration of Huanglian Ointment,and use the ELISA method to detect the changes in the levels of inflammatory chemokines after modeling and administration.RT-PCR method was used to detect the changes in the expression levels of JNK,ERK and ICAM-1 genes.Western Blot western blotting was used to investigate the changes in the expression levels of JNK,C-Jun,Jun-B,and FLG proteins.Result:(1)The best extraction process: directly put the medicinal materials into 5 times the amount of sesame oil,fry at 200℃for 4 hours.(2)Based on network pharmacology research methods,a total of 28 compounds were screened and 78 targets were selected;in PPI network analysis,there were 8 nodes with Degree ≥ 10,accounting for 10.3% of the total,including JUN,IL-6,MAPK1,MAPK14,STAT1,etc.Core target;79 items obtained through GO function enrichment analysis show that the Huanglian plaster treats eczema by regulating cytokine receptor binding(cytokine receptor binding),cytokine activity(cytokine activity)and other functions;obtained through pathway enrichment analysis 116 signaling pathways,pathways closely related to eczema:TNF signaling pathway,MAPK signaling pathway,IL-17 signaling pathway,Toll-like receptor signaling pathway,etc.There are complex interrelationships among these signaling pathways.(3)The Huanglian Ointment can improve dermatitis skin damage,inflammatory cell infiltration and redness induced by Biostir-AD ointment,repair the damaged skin barrier,and is positively correlated with the dosage.Compared with the model that only applied Biostir-AD ointment,the Huanglian ointment reduced the secretion of inflammatory factors TNF-a,IFN-r,IL-6,Ig E induced by dust mites,and inhibited the development of the body’s inflammatory response;The results of immunohistochemistry showed that the Huanglian Ointment increased the expression levels of C-Jun and FLG proteins and promoted skin cell proliferation,which proved that Huanglian Ointment has a good effect on the treatment of eczema.(4)The Huanglian Ointment oil can reduce the secretion of inflammatory chemokines in Hacat cells and significantly inhibit the release of inflammatory mediators;by regulating the JNK and ERK pathways,reducing the expression of ICAM-1 and reducing the inflammatory response of epidermal cells;promoting the proliferation of skin cells And differentiation,restore the balance of C-Jun and Jun-B,increase the secretion of Flg protein,and play a role in repairing the skin barrier.Conclusion:(1)The optimal extraction process is highly repeatable,stable and feasible,and has practical value.It standardizes the operation of the ancient recipe technology to "pour the medicine with dryness",and lays a solid foundation for the mass production of Huanglian Ointment.(2)The Huanglian Ointment can significantly improve the skin damage of guinea pig eczema induced by dust mites.(3)The Huanglian Ointment has a good anti-inflammatory effect on Hacat cell inflammatory response stimulated by TNF-a and IFN-r,and it has a positive correlation with the dosage.(4)The Huanglian Ointment oil inhibits the JNK signaling pathway,reduces the secretion of inflammatory chemokines,increases the expression of C-Jun,promotes cell proliferation,and repairs the damaged skin barrier.