Paeonol Interferes With The Regulation Of SOCS1-STAT3 By MicroRNA-155 In HaCaT Cells Of Psoriasis

This dissertation is composed of two parts:literature review and experimental research.The part of literature review:The first literature review:a review of recent advances in research on the treatment of blood-hot psoriasis has found that many physicians have used hot and cold blood prescriptions as the core of treatment,both in clinical research and experimental research.In recent years,the main research direction has been greatly studied from the traditional Chinese medicine prescription to the traditional Chinese medicine monomer,and the research on the monomer of the traditional Chinese medicine trying to clarify the mechanism of Chinese medicine treatment is still a hot research direction in recent years,so this is used as one of the basis for this experimental study.The second literature review:SOCS1(Suppressors of cytokine signaling-1)as a cytokine inhibitor has the effect of negative feedback regulating signal pathway activation.There have been studies showing that microRNA-155 is highly expressed in psoriasis and can directly act on the SOCS1 gene,while SOCS1,STAT3 and other factors are highly specific in psoriasis.Expression,Therefore,we speculate that microRNA-155 may regulate the development of silver chip disease by regulating the expression of SOCS1 and related signal molecules,reducing the secretion of pre-inflammatory cytokines,inhibiting local inflammation and excessive proliferation of keratinoblasts.This is also the foundation of this experimental study.The part of experimental research:Objective:To investigate whether paeonol regulates the expression of SOCS1-STAT3 by intervening miR155in the psoriasis model of HaCat cell.Mehtods:Hcact cells were molded by cytokine stimulation after transfection of Pre-/anti-miR-155,and the drug serum was used for intervention.Divided into the following 6 groups:Group A paeonol intervention group(HaCat cells were stimulated by cytokines and then replaced with culture medium),Group B anit-miR-155 group(HaCat cells were stimulated by cytokines after transfection of anti-miR-155),Group C pre-miR-155 group(HaCat cells were stimulated by cytokines after transfection of Pre-miR-155),Group D transfection control group(HaCat cells were stimulated by cy-tokines after transfection of control RNAi),Group E blank control group(HaCat cells were stimulated by cytokines and then replaced with normal medium),Group F pre-miR-155 + paeonol interrvention group(HaCat cells were stimulated by cytokines after transfection of Pre-miR-155 and then replaced with culture medium).The expression of SOCS1 and STAT 3 between different groups was measured and compared by PCR and Western Blot.Results:(1)The PCR method detected SOCS1 expression:Group E compared with Group D(P>0.05),Compared with Group E,the expression level of SCOS1 increased(P<0.05),group C and group D compare the low level of SOCS1 gene expression(P<0.05),Group B compared with Group C,showed that transfection intervention and drug intervention were m eaningful,and that over-expression and inhibition of expression miR-155 caused the expression of SOCS1 gene to decrease and increase accordingly.Group F compared with group C to increase SOCS1 expression level(P<0.05),indicating that tannin may play a role similar to inhibiting miR-155 expression;(2)PCR method to detect STAT3 expression:There was no statistical difference between E group and D group(P>0.05)The average water expression of Group B and Group C was higher than that of Group D STAT3,and there was a statistical difference(P<0.05)Compared with Group A,Group C,Group D,and Group F,the expression level of STAT3 decreased and there was a statistical difference(P<0.05),indicating that cells that have been transfected or interfered with by tannin have no special significance for the expression of STAT3;(3)The Western blot method tested the protein expression of SOCS1,STAT3,and p-STAT3 in each group and detected specific positive bands,indicating that the experimental operation was normal and an ideal band distribution was obtained;With the Group E as a reference,the SOCS1 expression level in Group C was low,while the SOCS1 expression levels in Groups A and F were high,and Group F expressed more than the SOCS1 protein in Group C,which was consistent with our PCR results,while each group STAT3 had expression.There is little difference in the expression of each group,but the expression of p-STAT3is not the same.Group E is the reference.Groups A,B,C,and F all have different levels of P-STAT3 expression.Group B has the highest expression.Group F was higher than group C’s p-STAT3,indicating that STAT3 and p-STAT3 may have a state of dynamic equilibrium,and Danpiphol has no significant interference with it.Conclusion:(1)Paeonol acts as the equivalent of anti-miR-155 expression,which may act as a negative adjustment to SOCS1 by inhibiting miR-155 expression.(2)Paeonol has no significant effect on STAT3.