The Impact Of Opioids On The Pmn Of Nf-{kappa} B Gene And Apoptosis Of Pmn

Objective 1. To determine the effect of morphine and fentanyl on expressions of subunit of nuclear factor-KB(NF-κKB)—p65 mRNA and content of p65 protein in human polymorphoneclear neutrophils(PMN) in vitro with or without LPS; 2. To observe the influence of exogenous opioids and fentanyl on apoptosis of human polymorphoneclear neutrophils(PMN) in vitro; In order to evaluate the immunomodulatory effect of opioid to patients who received analgesia after operation , especially to critically ill patients . Methods PMN were isolated from fresh whole blood healthy human volunteers by using Annmarie L method.and were resuspended in RPMI-1640 media with concentration of 1×10¹⁰L^-1 and seeded in plates (1×10⁹L^-1 per well). Subsequently, cells were incubated in media containing buffer(control, groupC), LPS(100ng/ml, groupLPS), morphine(50nmol/L, groupM), fentanyl(100nmol/L, groupF), morphine50 nmol/L+ LPS 100ng/ml (groupM+LPS) , fentanyl 100nmol/L+LPS100ng/ml (groupF+LPS) respectively for 3h. Reverse transcription-polymerase chain reaction(RT-PCR) was used to detect mRNA expressions of p65 , a specific antibody against the p65 subunit of NF-κB was used and the content of p65 protein was detected by Western blotting.Neutrophils from other 22 healthy volunteers were isolated, divided into 6 groups, given LPS morphine and fentanyl, and incubated for 12h and 24h respectively, the methods of isolation and incubation of PMN were carried out by the same way as that described above. The percentage of apoptotic neutrophils was assessed by immunofluorescence flow cytometry of Annexin V and propidoum iodide(PI) staining.All data were present as the mean±SEMo Deferences between the various treatments were assessed for statistical significance by measures ANOVA,followed by comparison test. A probability value less than .05 was considered significant. SPSS11.0 was used for all statistical analyses. Results1. The results of RT-PCR indicated that LPS significantly increased the expression of p65mRNA (group LPS 2.166±0.105 vs group C 1.643±0.086 , p<0.01) ; While...