The Mechanism Of Worenine Suppresses Psoriasis By Targeting Aurora-B

Background: Psoriasis is a common immune-mediated chronic inflammatory skin disease with a global prevalence of about 2%.It can affect the skin,nails and joints of patients.Studies have shown that psoriasis can affect a person’s self-esteem,increase the risk of heart disease,obesity,diabetes,arthritis,depression and other diseases,and thus reduce the quality of life.Psoriasis is prone to relapse and prolonged,and its complete cure is the main problem at present.The solution of this problem is conducive to improving the quality of life for patients and reducing the risk of other diseases,which has great clinical value and social significance.Traditional Chinese medicine rhizome coptidis has anti-inflammatory effect.Worenine is a monomer component extracted from rhizome coptidis,and its effect and mechanism in psoriasis are unknown.Objective: To investigate the mechanism of Worenine suppressing psoriasis skin.Methods: 1.A certain concentration of M5(TNF-α,IL-17 A,IL-22,IL-1α and Oncostatin-M)was added to human keratinocytes Ha Ca T and treated at different times.Cells were collected and total protein was extracted.2.MTT was used to detect the toxic effect of Worenine on cells,and appropriate concentration was selected for subsequent tests.3.Worenine was added to the psoriasis-like cell model,the expression level of psoriasis marker protein S100A7 and the protein level and phosphorylation level of STAT3 were detected by Western Blot.4.Kinase Array was used to screen potential targets of Worenine.5.In vitro kinase experiment was carried out to verify whether Worenine can inhibit Aurora-B in vitro.6.Worenine was added to the psoriasislike cell model,and the phosphorylation level of Histone-H3(Ser10),which can reflect the Aurora-B activity,was detected by Western Blot.7.Established the mouse model of psoriasis,applied carbolic 940,Worenine or Hydrocortisone(HYD)to the backs and ears of mice,and then applied Imiquimod(IMQ)for 5 days,finally killed the mice.8.During animal modeling,morphological changes of mice’s backs and ears were observed and photographed every day,and the psoriasis area and severity index(Psoriasis area and severity index,PASI)was used to evaluate the severity of Psoriasis.9.The skin of mice’s backs and ears was stained with H&E,and the pathological changes were noted.10.S100A7,STAT3,p-STAT3,p-Aurora-B and p-Histone-H3 were detected by Western Blot after homogenizing the skin on the back of mice,and the effect of Worenine on STAT3 and Aurora-B activity was further detected.Results: 1.After addition of M5 to Ha Ca T,the expression level of psoriasis marker protein S100A7,the protein level and phosphorylation level of STAT3 increased,and the psoriasis-like cell model was established.2.Worenine was added to Ha Ca T,and the concentration of cell activity was above 80% and concentration gradients(0,2 μM,5 μM and 12.5 μM)were set for subsequent experiments.3.After Worenine was added to the psoriasis-like cell model,the expression level of psoriasis marker protein S100A7,the protein level and phosphorylation level of STAT3 were decreased,confirming that Worenine could inhibit psoriasis in Ha Ca T cells in a concentration and time-dependent manner.4.Kinase Array assay identified Aurora-B is a potential target of Worenine.5.In vitro kinase experiment confirmed that Worenine could inhibit the kinase activity of Aurora-B in vitro.6.P-Histone-H3(Ser10)was reduced after Worenine was added to the psoriasis-like cell model,confirming that Worenine could inhibit Aurora-B kinase activity at the cellular level.7.Psoriasis-like animal model induced by IMQ was successfully established.8.In the animal model,comparing with the IMQ group,the psoriatic lesions in the back and ear skin of mice in the Worenine group were significantly alleviated,and the PASI index was significantly decreased,indicating that Worenine could inhibit psoriasis in morphological and pathological studies.9.S100A7,STAT3,p-STAT3,p-Aurora-B and p-Histone-H3(Ser10)were decreased in the back skin of mice,further confirming that Worenine could inhibit the development of psoriasis by inhibiting the activity of STAT3 and Aurora-B.Conclusion: 1.Worenine could inhibit the occurrence of psoriasis at the cellular and animal levels.2.Worenine could inhibit the activation of STAT3 caused by psoriasis.3.Aurora-B is a potential target of Worenine.4.As a potential inhibitor of Aurora-B,Worenine could relieve the symptoms of psoriasis,providing a new direction and target for the treatment of psoriasis.