Xiaoqinglong Decoction Based On TSLP-stimulated DCs Was Used To Study The Mechanism Of Th2 Treatment Of AR Cells

ObjectiveStudies have suggest that(TSLP)can activate and change the characteristics of dendritic cells(DCS)to form a microenvironment which is conducive to the differentiation of Th2 induing the occurrence of allergic rhinitis.Minor Green-Blue Dragon Decoction,an traditional Chinese medicine,has a good effect in the treatment of allergic rhinitis whereas the specific mechanism of anti-allergic effect is not clear.This experiment was carried out on the TSLP-activated DCs system and intervened by Minor Green-Blue Dragon Decoction to observe whether the Minor Green-Blue Dragon Decoction can prevent allergic rhinitis in TSLP activated DCs systemMethods1.To establish the method about culture of DCs by GM-CSF and IL-4 in vitro.2.To preparae the water extract from Minor Green-Blue Dragon Decoction and to detect the effect of Minor Green-Blue Dragon Decoction on DCs by CCK-8.3.To select the optimal dose of TSLP on DCs by detecting the expression of CD83 and IL-13.4.To detectthe purity of DCs and the expression of CD83 after intervention of Minor Green-Blue Dragon Decoction on TSLP activated DCs in vitro by FCM.5.To detect the expression of p-JAK1?p-JAK3?p-STAT5?p-STAT6 and OX40L after intervention of Minor Green-Blue Dragon Decoction on TSLP activated DCs in vitro by Western blot.6.To detect the expression of CCL17mRNA?CCL22Mrna after intervention of MinorGreen-Blue Dragon Decoction on TSLP activated DCs in vitro by RT-PCR.7.To establish the method of co-culture about DCs and T cell in vitro.8.To detect the expression of GATA-3,C-Maf,and T-bet after intervention of Minor Green-Blue Dragon Decoction on TSLP-activated DCs-ThO in vitro by RT-PCR.Results1.Bone marrow cells and spleen cells can be cultured in vitro to produce DCs with better performance.2.The better concentration range of Minor Green-Blue Dragon Decoction on DCs was screened.3.The expression of CD86 and IL-13 was the highest in lOng/ml TSLP group and there was significant difference compared with the control group.4.TSLPR could significantly decrease the expression of OX40L,CCL17,CCL22,and so on compared with TSLP group(P<0.05).5.4ug/ml Minor Green-Blue Dragon Decoction can reduce significantly the expression of OX40L at the protein and gene level compared with TSLP group(all P<0.05).6.2ug/ml,4ug/ml,and 8ug/ml Minor Green-Blue Dragon Decoction can reduce significantly the expression of TARC/CCL17 and MDC/CCL22 compared with TSLP group(all P<0.01).7.TSLP has no obvious effect on the expression of IP-10 compared with the control group.8.Minor Green-Blue Dragon Decoction has no obvious effect on the expression of IP-10 compared with TSLP group.9.2ug/ml,4ug/ml,and 8ug/ml Minor Green-Blue Dragon Decoction can reduce significantly the expression of CCL17mRNA compared with TSLP group(all P<0.01).10.4ug/ml and 8ug/ml Minor Green-Blue Dragon Decoction can reduce significantly theexpression of CCL22mRNA compared with TSLP group(all P<0.01).11.2ug/ml,4ug/ml,and 8ug/ml Minor Green-Blue Dragon Decoction can reduce significantly the expression of GATA-3 compared with TSLP group(all P<0.01).12.4ug/ml and 8ug/ml Minor Green-Blue Dragon Decoction can reduce significantly the expression of C-Maf compared with TSLP group(P<0.01,P<0.05).13.TSLP has no obvious effect about the expression of T-bet.14.2ug/ml and 8ug/ml Minor Green-Blue Dragon Decoction piay a certain role in promoting the expression of T-bet compared with TSLP group(P<0.01).Conclusions1.DCs with better purity and performance can be obtained from Bone marrow cells and spleen cells in vitro.2.TSLP can activate DCs to form a microenvironment which is conducive to the differentiation of Th2 induing the occurrence of allergic rhinitis.3.TSLPR plays a certain role in blocking the activation of DCs caused by TSLP.4.Minor Green-Blue Dragon Decoction can prevent allergic rhinitis by affecting the expression of CCL17,CCL22,OX40L,and JAK/STAT in TSLP-activated DCs system.5.Minor Green-Blue Dragon Decoction can prevent allergic rhinitis by reducing the expression of GATA-3 and C-Maf in TSLP-activated DCs-ThO system.6.Minor Green-Blue Dragon Decoction play an anti-allergic effect by increasing the expression of T-bet in TSLP-activated DCs-ThO system.7.Minor Green-Blue Dragon Decoction has no obvious effect on the expression of IP-10.