Effects Of Pingchuan Granule On Regulation Of Dendritic Cells Function In Asthmatic Rats Via NOTCH Singnaling

Objective:To observe the effects of Pingchuan Granule on regulation of dendritic cells function in asthmatic rats,and elucidate the mechanism of Pingchuan Particles on chronic asthma.Methods:1.40 Wistar rats were randomly divided into four groups:control group,model group,dexamethasone group and Pingchuan Granule group.Bronchial asthma rats model was established using the ovalbumin excitation method,except the control group.Rats in Pingchuan Granule group were treated with Pingchuan Granule(5.4g/kg).Rats in the dexamethasone group were treated with dexamethasone(0.45mg/kg),Rats in control group and model group were treated with 0.9%NaC1(5ml/kg).Lung tissues,bronchoalveolar lavage fluid were collected,lung pathological changes and number of DCs in lung tissues were observed after hematoxylin and eosin staining.Lung DCs were isolated from lung tissues.Expression of CD80,CD86 and MHCIIwere analyzed using flow cytometry.Expression of Notch and Jag-1 mRNA were determined by Realtime-PCR.Expression of EGR1 and PKC protien were determined by Western blot.2.Rats bone marrow cells were cultuered with rmGM-CSF(10ng/ml)?rmIL-4(10ng/ml))for 7 days,and then got DCS?To study the effect of LPS on DCs function.DCs were treated with increasing concentrations of LPS(0.1,0.5,1,1.5,2?g/ml)for 3,6,12,24hours respectively.To study the effect of DAPT on DCs.DCs were treated with increasing concentrations of DAPT(0,1,5,10,20,50 ?mol/L)for 48 hours respectively.DCs were divided into five groups:control group,LPS group,DAPT+LPS group,Pingchuan Granule+LPS group,and rat serum +LPS group.DCs in LPS group were cultuered with LPS(1.5?g/ml).DCs in DAPT+LPS group were cultuered with DAPT(20?mol/L)and LPS(1.5?g/ml).DCs in Pingchuan Granule+LPS group were cultuered with Pingchuan Granule and LPS(1.5?g/ml).DCs in rat serum +LPS group were cultuered with rat serum and LPS(1.5?g/ml).Expression of CD80,CD86 and MHCIIwere analyzed using flow cytometry.Expression of Notch and Jag-1 mRNA were determined by Realtime-PCR.Expression of EGR1 and PKC protien were determined by Western blot.Results:1.After the treatment,the inflammatory in lung tissue was improved,and the number of DCs in lung tissue was decreased in the Pingchuan Granule group.(P<0.05 compared with the model group),and there was no significant difference compared with the dexamethasone group(P>0.05).2.The expression of CD80,CD86 and MHCII in lung DCs of Pingchuan Granule group,were 62.04±4.57%,54.66±4.40%and 57.64±4.26%,respectively.And there was significant difference compared with the model group(all P values<0.05).3.The expression of Notchl and Jag-1 mRNA were decreased in the Pingchuan Granule group.There was significant difference compared with the model group(all P values<0.05).4.The expression of PKC protein was decreased in the Pingchuan Granule group,and the expression of EGR1 was increased.There was significant difference compared with the model group(all P values<0.05).5.These results indicated that LPS could improve the development of DCs.The improvement was in a concentration-dependent and time-depent manner.The greatest effect observed at a concentration of 1.5?g/ml for 12 hours,which had no difference with 2.5?g/ml group,in stimulating DCs maturation and cell inhibition(P>0.05).6.DAPT could inhibit NICD expression,and the inhibition was in a concentration-dependent manner with the greatest effect observed at a concentration of 20 p,mol/L(P values<0.05,compared with other groups).7.The expression of CD80,CD86 and MHCII induced by LPS,were decreased Pingchuan Granule group(all P values<0.05).The expression rates were 62.26±4.44%.67.69±5.33%and 63.77±4.58%,respectively.8.The expression of Notchl and Jag-1 mRNA induced by LPS,were decreased in Pingchuan Granule group(all P values<0.05).9.The expression of PKC protein induced by LPS,was decreased in the Pingchuan Granule group,and the expression of EGR1 was increased.There was significant difference compared with LPS group(all P values<0.05).Conclusions:1.Pingchuan Granule can improve lung pathological changes in asthmatic rats.2.Pingchuan Granule can inhibite DCs maturation in asthmatic rats.3.Pingchuan Granule can reduce the expression of Notchl and Jag-1 mRNA in DCs.4.Pingchuan Granule may regulate the immune function of DCs,by inhibiting the expression of DCs Notchl mRNA.