Effect Of The Monomers Of Chinese Medicine Of Bushen Yijing Therapy On Endothelial To Mesenchymal Transition In Systemic Scleroderma

Objective1.To summarize the theoretical basis and clinical application of Bushen Yijing herbal compound in the treatment of systemic scleroderma.2.To observe the effects of three monomers,ursolic acid(UA),acteosid e(AC)and hydroxy safflower yellow pigment A(HSYA)of Chinese herbas comp ound in Bushen Yijing Therapy on endothelial to mesenchymal transition(E ndMT)in systemic scleroderma(SSc),In thus to provide a new basis for the clinical application of Bushen Yijing Therapy,as well as to provide new ideas for the treatment of SSc and the development of New drug.MethodsPart 1:Clinical researchBased on the theory of the Correlation of TCM Lung,spleen kidney,We summarize the characteristics of Bushen Yijing Therapy in clinical application in SSc through literature collection and analysis.Two case reports of SSc,which were treated with Bushen Yijing Therapy,were used to demonstrat the effects of Bushen Yijing Therapy in SSc,as well as to provide the theoretical basis for the further experiments.Part 2:Experiment researchHUVEC cells were induced by TGF-β 1 to establish the EndMT model of SSc,and the model was treated with three monomers,UA,AC and HSYA respectively at 24 and 48h,and then detected the mRNA expression of CD31,FSP1,vimentin by QPCR,and the relative protein expression were checked by WB.Results1.Clinical observations found that the prescription of Bushen Yijing Therapy can improve the clinical symptoms and the condition in SSc.2.The cells in EndMT model were changed after induced by TGF-β1.The QPCR showed that the CD31mRNA decreased and the FSP1,Vimentin mRN A increased significantly in EndMT model,the EndMT models were successfully established.3.CD31mRNA in UA24h groups and UA48h groups were both significantly higher(P<0.05,P<0.05),and FSP1 mRNA decreased significantly(P<0.05,P<0.05),Vimentin mRNA decreased significantly as well(P<0.05,P<0.05)than the model groups.The decrease of FSP1,Vimentin mRNA in UA48h groups were more obvious than that in UA48h group(P<0.05,P<0.05).There was no significant difference in CD31mRNA between UA24h group and UA48h groups.WB showed that the expression of CD31 protein were decreased,and the expression of N-cadherin、Vimentin protein were significantly increased in the model groups.In the two UA groups,the expression of CD31 protein were both increased,the expression of N-cadherin and Vimentin protein were decreased.The expression CD31 protein of UA 24h groups were as same as that in UA48h groups,and the expression of N-cadherin and Vimentin protein in UA48h groups were both lower than that in UA24h groups.4.The expression of CD31 mRNA in AC24h groups and AC48h groups were both significantly higher(P<0.05,P<0.05),and FSP1 mRNA decreased significantly(P<0.05,P<0.05),Vimentin mRNA decreased significantly as well(P<0.05,P<0.05)than the model groups.There were no significant difference in CD31,Vimentin mRNA between AC24h groups and AC24h groups(P>0.05,P>0.05).The expression of FSP1 mRNA in AC 24h groups were lower than that in AC48h groups(P<0.05).CD31 protein in AC two groups were slightly higher.The N-cadherin protein decreased in different degree in AC groups,as the AC48h groups decreased more significantly.Vimentin protein in AC24h groups was lower than that in model groups,while AC48h groups were slightly higher.5.The expression of CD31 mRNA in HSYA24h groups and HSYA48h groups w ere both significantly higher(P<0.05,P<0.05),and FSP1 mRNA decreased significantly(P<0.05,P<0.05),Vimentin mRNA decreased significantly as wel 1(P<0.05,P<0.05)than the model groups.CD31 mRNA increased more significan tly(P<0.05)and Vimentin mRNA decreased more significantly in HSYA 48h gr oups(P<0.05)in HSYA48h groups.The CD31,E-cdherin protein decreased and Ncadherin protein increased in the model groups.Compared with model group s,CD31 protein in HSYA24h groups were lower,HSYA 48h group were slightly higher.The E-cadherin protein of HSYA groups were higher than model grou ps,HSYA 48h groups were higher obviously.There was no difference of N-cad herin protein between HSYA groups and model groups.Conclusion1.Applying the prescription of Bushen Yijing Therapy can improve the clinical symptoms in SSc patients.2.The three monomers of UA,AC and HYSA can increase the expression of endothelial interstitial markers and decrease the expression of interstitial markers,pointing out that they may inhibite the EndMT of scleroderma and can be used for further study.