| Objective To explore the therapeutic effect and mechanism of Psori-CM02 on psoriasis by vivo and vitro experiments.Methods Four classic psoriasis animal models were conducted to evaluate the therapeutic effect In vivo experiment.For example :(1)To investigate the therapeutic effect of drugs on Psoriasis like model in Guinea pigs by propranolol hydrochloride.(2)To investigate the therapeutic effect of drugs on Psoriasis like model in mice by imiquimod.(3)To investigate the therapeutic effect of pruritus on Itching models in mice by Dextran 40.(4)Tail skin scales granular layer model in mice investigate the therapeutic effect of drugs on psoriasis skin parakeratosis.(5)Mouse vaginal epithelial mitosis inhibition experiment to investigate inhibition of drug on cell proliferation.(6)Granuloma model in rats by cotton ball to investigate the therapeutic effect of drug on chronic inflammation.(7)Blood stasis rats model by Epinephrine combined with ice bath to investigate drug promoting blood circulation to remove blood stasis.Four aspects in vitro experiment around the protection of drugs for cell proliferation,apoptosis accelerant,effectiveness for MAPK signaling pathways.For example:(8)MTT method to investigate the proliferation inhibitory of Psori-CM02 on Ha Cat,HUVEC,KB cell in vitro.(9)Flow cytometry method to investigate effects of Psori-CM02 on Ha Cat cell apoptosis.(10)MTT method to explore the protection of Psori-CM02 on RAW264.7 cell by LPS from inflammation.(11)NO Elisa kits to investigate effect of Psori-CM02 on NO releasing of RAW264.7 induced by LPS.(12)Western blot method to investigate effect of Psori-CM02 on the MAPK pathway of RAW264.7 cells induced by LPS.Results(1)The results of psoriasis model in guinea pig by Propranolol and pathologic suggest that Psori-CM02 all the treatment groups can improve the psoriasis,no dose difference.(2)The results of psoriasis mice model by imiquimod suggest that Psori-CM02 can reduce the PSAI,ki67 index in epidermis cell and IL-17 level in serum contrast with model,p<0.05.The pathological results suggest three dose of the reagent is better than the model group.(3)In mouse vaginal epithelial mitosis inhibition experiment,the results show that the treatment groups can effectively inhibit mitosis.Inhibition rate of Yu Jin,MTX group and the low dose group,middle dose group and high dose group respectively for 18.3%,21.9%,15.5%,18.3% and 30.6%.(4)In mouse tail test,the results showed that all groups can increase the ratio of granular layer in the scales,contrast with the model group,differences have statistical significance,p < 0.05.(5)The results of body itch model in mice by Dextran 40 indicate that all the treatment groups can effectively restrain the mice body itching,frequency of itchy in 30 minutes less than model group,difference has statistical significance,p < 0.05.(6)The results of granuloma model in rats by cotton ball indicate that dry weight of granuloma in Model group is 77.6±15.4 mg more than indomethacin group: 46.4±12.2 mg,middle dose group:61.8±11.4 mg and high dose group:60.8±2.5 mg,difference have statistical significance,p < 0.05.(7)The results of acute blood stasis model in rat indicate that whole blood viscosity of 3 different dose groups(in addition to low doses of 30 s-1)of Psori-CM02 are lower than the model group,the difference had statistical significance,P < 0.05.Erythrocyte aggregation index in each treatment group is less than the model group,the difference had statistical significance,p<0.05.(8)MTT results indicate that Psori-CM02 have obvious inhibitory effect on Ha Ca T cells and HUVEC cells,no significant inhibitory effect on the KB cells.(9)Flow cytometry results showed that the concentration of 0 ?g/m L,1 ?g/m L,10?g/m L of Psori-CM02 in Ha Ca T,24 h,the apoptosis percentage were 2.63%,4.48% and2.63% respectively;48 hours,the apoptotic percentage were 2.80%,3.85%,13.63%,respectively,of which 10 ?g/m L concentration was more effective on Ha Ca T cell apoptosis.(10)Psori-CM02 can protect RAW264.7 from the inflammation induced by LPS.There are concentration-response relationship apparently.(11)NO has not been measured in the control group.LPS stimulation group released the most NO,while the Psori-CM02 treatment group reduce in turn with the concentration gradient increase.(12)Psori-CM02 can inhibit the phosphorylation of p38,Erk1/2 and JNK in the concentration of 1-200 ?g/m L,which suggest Psori-CM02 may participate in blocking the MAPK signaling pathways on RAW246.7 induced by LPS.Conclusion In vivo experiment,results indicate Psori-CM02 can improve Symptom of psoriasis on the animal model.Psori-CM02 can inhibit mitosis of epithelial basal cell,increase granular layer of the mouse tail squama,reduce itching and chronic inflammation,promote blood circulation to remove blood stasis.Vitro experiments showed that drugs can inhibit Ha Ca T cell proliferation,increase the rate of apoptosis and differentiation,also can block Raw264.7 cells MAPK signaling pathways,reduce the NO product to improve inflammation.In conclusion,Psori-CM02 treat psoriasis through multiple ways and targets.The treatment reflects the characteristics of traditional Chinese medicine fully. |
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