Experimental Study On The Inflammatory Effect Of Mongolian Medicine Sendeng Four-flavor Soup On Mice With Psoriasis

Objective: To explore the therapeutic effect and mechanism of Mongolian medicine Sendeng four-flavor soup on mouse model of psoriasis induced by imiquimod cream(IMQ),and to provide theoretical basis for further promotion and application of Mongolian medicine Sendeng four-flavor soup in the treatment of psoriasis.Methods:The Mongolian medicine Sendeng four-flavor soup was extracted with 95% ethanol and made into freeze-dried powder by freeze-drying technology.Select 8 weeks BABL/c male mice,randomly divided into 6 groups,each group of 10,the blank control group(CK),MODEL group(MODEL group),methotrexate(MTX)group,Sendeng’s low dose group(Sendeng’s L group),Sendeng’s Medium dose group(Sendeng’s M group),Sendeng’s high dose group(Sendeng’s H group).CK group apply 62.5mg of Vaseline cream daily on the back,The other five groups were daubed with 62.5mg5%IMQ cream on the back daily,for 10 days in a row.CK group,MTX group,Sendeng’s L group,Sendeng’s M group and Sendeng’s H group were given normal saline,MTX(2mg/kg),low-dose Mongolian medicine Sendeng four-flavor soup(0.315g/kg),medium-dose Mongolian medicine Sendeng four-flavor soup(0.63g/kg)and high-dose Mongolian medicine Sendeng four-flavor soup(1.26g/kg)0.2ml/d by intragastric administration from the 4th day for consecutive 7 days.Psoriasis lesion area and severity index score(PASI score)were calculated to evaluate the severity of lesions.Histopathological changes were detected by HE staining of tissue sections,and proliferation was detected by Ki67 immunohistochemistry.Spleen index was calculated,spleen index = spleen weight(mg)/body weight(g);Flow cytometry was used to detect the ratio of Th17/Treg cells in inguinal lymph nodes.The expressions of p-p38、p38、p-STAT3、STAT3、p-JAK1 and JAK1 pathway proteins in skin lesions were detected by western blot.Results:(1).Compared with CK group,mice in MODEL group showed significant skin hypertrophy,erythema and scales;Compared with the MODEL group,at the same time,Sendeng’s L,M,H and MTX groups showed reduced erythema,scarier scales,reduced skin thickness,and decreased PASI scores.MTX group and Sendeng’s M group had the most obvious effect,and Sendeng’s L group and Sendeng’s H group followed.(2).HE staining of tissue sections and Ki67 immunohistochemistry: Compared with CK group,mice in MODEL group showed thickening of epidermal cuticle,parakeratosis,Munro microabscess,dermal capillary hyperplasia and lymphocyte infiltration,with high expression of Ki67;Compared with the MODEL group,the corneous layer thickness and parakeratosis in the Sendeng’s M group and the MTX group were reduced,Munro microabscess was reduced or even disappeared,dermal capillary hyperplasia and lymphocyte infiltration were reduced,and Ki67 expression was lower.Sendeng’s L group and Sendeng’s H group can also improve the pathological structure of mice,but the degree is not as good as Sendeng’s M group.(3).The spleen index of the MODEL group was about three times higher than that of the CK group.After treatment,the spleen index of the MTX group and the Sendeng’s M group was reduced by half compared with that of the MODEL group;Compared with MODEL group,spleen index of Sendeng’s L group had no significant change,while spleen index of Sendeng’s H group decreased,but the decline degree was less than that of Sendeng’s M group.(4).Flow Cytometry: Compared with CK,the proportion of Treg cells was decreased and the proportion of Th17 cells was increased in MODEL group;Compared with the MODEL group,the proportion of Treg cells increased in MTX group,Sendeng’s M group,Sendeng’s L group and Sendeng’s H group,while the proportion of Th17 cells decreased,and the effect of MTX group and Sendeng’s M group was the most obvious,followed by Sendeng’s L group and Sendeng’s H group.(5).western blot: Compared with CK group,the expressions of p-p38,p38,p-STAT3 and pJAK1 were increased in MODEL group;Compared with the MODEL group,the expressions of p-p38,p38,p-STAT3 and p-JAK1 were significantly decreased in the MTX group and the Sendeng’s M group,and the expressions of p-p38,p38,p-STAT3 and pJAK1 were also decreased in the Sendeng’s L group and the Sendeng’s H group,but to a lesser extent than that in the Sendeng’s M group,and there was no significant difference in the expressions of STAT3 and JAK1 among all groups.Conclusion:(1).Mongolian medicine Sendeng four-flavor soup can effectively relieve the symptoms of psoriasis mouse dermatitis induced by IMQ,reduce PASI score,improve pathological structure characteristics,reduce the expression of Ki67,and lower spleen index.The optimal concentration is medium dose(0.63g/kg);(2).Mongolian medicine Sendeng four-flavor soup has an inhibitory effect on imq-induced psoriatic-like mice,which is related to the regulation of Th17/Treg cells(up-regulation of Treg cells and down-regulation of Th17 cells),and the reduction of p38,p38,Stat3 and JAK1 phosphorylation levels,and may play a role through JAK-STAT and p38 MAPK protein pathways.