The Expression Of TNF-α,NF-κB Induced By Norepinepherine In Cultured Neonatal Rat Cardiomyocytes And The Intervention Effects Of Morphine And Tramadol

Objective The aim of the study was to investigate the effects of morphine and tramadol preconditioning on TNF-α,NF-κB expression induced by norepinepherine in cultured neonatal rat cardiomyocytes to explore the mechanism of myocardial protective effects of morphine and tramadol.Methods (1)The model of original cultured cardiomyocytes of SD rat was established and to observe and identify neonatal rat cardiomyocytes with morphology and immunocytochemical assay method under a reverse microscope. ( 2 ) 40 holes rat cardiomyocytes were randomly divided into 10 groups: the control group; 3h group; 6h group; 12h group; 24h group; NE1 group(10-6 mol·L^-1); NE2 group(10-7mol·L^-1); NE3 group(10-8 mol·L^-1) ; MP group; TMD group. Cells were pre-administrated 10-5mol·L^-1 MOR,TMD before NE 30 minutes, respectively. The cells was cultured as scheduled to process for TNF-αenzyme immunometric assay (EIA) ,for TNF-αmRNA Semi-quantitative examination using RT-PCR technique,for TNF-αimmunocyto- chemistry in order to observe the change of TNF-αinduced by norepinepherine. (3)32 holes rat cardiomyocytes were randomly divided into 8 groups: the control group; NE1 group(10-6 mol·L^-1); NE2 group(10-7mol·L^-1); NE3 group(10-8 mol·L^-1) ; MP group; TMD group;Ph group;Es group. Cells were pre-administrated 10-5mol·L^-1 MOR,TMD,Ph,Es before NE 30 minutes, respectively.The cells was cultured as scheduled to process for NF-κB using Flow Cytometry (FCM) technique,for NF-κB immunocyto- chemistry in order to observe the change ofNF-κB induced by norepinepherine.Results (1)The results showed the differential changes in the levels of TNF-αand TNF-αmRNA in the NE group as compared with the baseline in the control group (p<0.05), the levels of TNF-αand TNF-αmRNA in cultured cardiomyocytes were significantly elevated after NE, and peaked in NE3 group,with 10-8 mol·L^-1NE for 24 hours. (2)The expression and activation of NF-κB increased significantly in NE group as compared with the baseline in the control group, the levels of NF-κB in cultured cardiomyocytes were significantly elevated after NE, and peaked in NE3 group,with 10-8 mol·L^-1NE for 24 hours.(3) The expression of TNF-αand TNF-αmRNA decreased significantly in MP group,TMD group as compared with NE3 group , but there was no significant differences among them. (4)The expression and activation of NF-κB decreased significantly in MP group,TMD group,Ph group,Es group as compared with NE3 group , but there was no significant differences among them.Conclusion (1) The NE could cause increase on TNF-αexpression,NF-κB activation in...