Study On The Expression And Function Of Progesterone Receptor In Peripheral Blood Polymorphonuclear Leukocyte Of Cattle

Progesterone(P)is one of the key hormones that induce the immunosuppression of bovine perinatal cells.The dynamic changes of the concentration of Progesterone play an important role in the distribution,quantity,status and function of polymorphonuclear neutrophil(PMN).But P-induced rabbit peripheral PMN signaling mechanism,especially receptor-mediated signal transduction mechanism remains to be explained.In this study,we taken the peripheral blood PMN of castrated cattle as the research object and used q PCR and western blot to investigate the effects of different concentrations of P on the gene transcription and expression of P receptor,membrane receptor,estrogen receptor and Immunologically active molecules TLR2,TLR4,NF-?B and so on in vitro and in vitro.That provide new evidence to determine the expression of P receptor in bovine PMN,reveal the characteristics of genomic and non-genomic signaling pathways that regulate PMN immune function in bovine peripheral blood,and elucidate the intrinsic relationship between the peripheral blood P concentration and the PMN immune activity.The test results show that:1.Receptor expression:(1)nPRs expression: bovine PMN expresses nPRs at gene transcription and protein level;low concentration P stimulates PRB expression,and increases the expression of Rb486 and LPS after addition of nuclear receptor inhibitor;high concentration P upregulates PRB expression RU486 had no significant effect on PRB expression,and PRB expression was down-regulated in LPS-induced inflammatory response.(2)mPRs(mPR?,mPR?,PGRMC1)expression: bovine PMN expressed mPRs at gene transcription and protein level;low concentration P down-regulated mPR? and PGRMC1 expression,up-regulated mPR? expression reached peak at 2h.The expression of mPR? and PGRMC1 were inhibited by high concentration P,and the inhibition of mPR? was inhibited and the inhibition was inhibited by RU486 and LPS.(3)ERs expression: low concentration of P upregulated ER? and GPR30,after adding RU486 showed a significant downward trend.2.Signaling of cellular active signaling pathways:(1)MyD88: high and low concentrations of P were raised MyD88,and reached a peaklevel after 30 min,low concentration of P in the role of RU486 inhibition of MyD88 expression,high concentrations of P no effect.(2)TLR2,TLR4: high and low concentrations of P were down-regulated TLR2,up-regulated TLR4 expression,and low concentrations of RU486 and LPS on the expression of both significant effect,all showed inhibitory effect.(3)NF-?B: high concentration of P inhibited NF-?B expression,RU486 and LPS inhibition of the role of the release;low concentration P after 30 min to lift the inhibition of NF-?B expression,RU486 and LPS on its expression Significantly affected.This study confirmed the presence of nPRs,mPRs and E receptors at the gene and protein levels of bovine peripheral PMN and acted as cross-mediated non-genomic effects of P.Progesterone exerts its antiinflammatory effect through PR-mediated genomic pathways and upregulates or downregulates membrane receptor-induced rapid non-genomic signaling;and some immunomodulatory effects may be mediated by upregulation of ERa and GPR30 rather than by PR signaling But still need further experiments to prove.