| Objective To investigate the intracellular calcium on the effect of pathogenesis of actue lung injury induced by paraquat intoxication and the protective and therapeutic effects of the inhibitor of Na~+/H~+exchanger-1 and Na~+ channels on the acute lung injury induced by paraquat poisoning ;Methods A549 cells were incubated with different concentration of paraquat and Amiloride Hydrochloride to establish experimental models .Half of cell survival rate detected with MTT assy to selected the median lethal concentration and median lethal time . 3 groups were divided according different drug application: PQ group exposed to paraquat with the concentration of 600μmol/L, Amiloride Hydrochloride group additionally exposed to Amiloride Hydrochloride with concentration of 20μmol/L,and blank control group without any drug intervention. Real time PCR was performed to analyze the expression level of NHE-1 mRNA in A549 cells of respectively groups.A549 cells were cultured in vitro and dyed with Fluo-3/AM, utilizing the advantage of real time monitoring the intracellular dynamic change of calcium by LCSM, the cells response and intracellular dynamic change of calcium level in A549 cells were detected after administration of PQ and Amiloride Hydrochloride. Data were expressed as mean±standard error of the mean((X|-)±S)statistical analysis was carried out with the soft SPSS 16.0Results The level of intracellular calcium increased after absorption of PQ(FI:1802.55±144.15 Vs 1470.90±225.71,P<0.01), then markedly decreased when given Amiloride Hydrochloride(FI:471.86±232.83 Vs 1802.55±144.15); Real time PCR showed that the NHE-1 gene expression level was significantly lower in Amiloride group than that in PQ group ( P<0.05).Conclusion Na~+/H~+exchanger-1 plays an important role in calcium overloaded induced by paraquat intoxication; Amiloride Hydrochloride can inhibit the increase of the level of intracellular calcium and protect against the PQ induced lung injury. |
PDF Full Text Request