Effects of neuroglia cultured supernatants with LPS-priming on neurons

Objective To study the effects of glia cultured supernatants with LPS-priming on neurons' viability and secretion activity.Methods Neurons and glia(mainly referring to astrocytes and microglia)were cultured in vitro and identified by immunofluorescence.Thereafter,these glial cells were divided into 4 groups:LPS-primed group,0.01μg/ml of LPS treated group,1μg/ml of LPS treated group and non-treated group.The LPS-primed group was preconditioned with 0.01μg/ml of LPS for 18 hours and re-retreated with a higher dose of LPS(1μg/ml LPS)for 24 hours.Then all cultured supernatants of 4 groups were collected and were used to treat the neurons for 24 hours in vitro.The cytotoxicity and secretion ability of proinflammatory cytokines were determined by CCK-8 kit and ELISA respectively.Results The cytotocixity of neurons was decreased by the treatment with the astrocyte supernatant of LPS-primed group,there was significant difference statistically if compared with 0.01μg/ml and 1ug/ml of LPS groups(P0.05;P0.05).The cytotocixity showed no profound variation after the treatment with the microglia supernatants of 4 groups(P0.05).As for TNF-α levels,it was increased by the treatment with the astrocyte supernatant of LPS-primed group,0.01μg/ml of LPS treated group,1μg/ml of LPS treated group,particularly in 1ug/ml of LPS treated group,statistical difference was found when compared with 0.01μg/ml of LPS treated group and non-treated group(P0.05;P0.05);IL-6 levels were increased after the treatment with the astrocyte supernatants of LPS-primed group,0.01μg/ml of LPS treated group,1ug/ml of LPS treated group(P0.05).Furthermore,after the treatment with the microglia supernatants of LPS-primed group,TNF-α level was increased profoundly when compared with non-treated group(P0.01).Though IL-6 levels exhibited an increasing trend,no significant difference was found in statistically between groups(P0.05).Conclusion There were diverse effects(ncluding protection or injury) of cultured supernatants from astrocytes and microglia on the cytotoxicity and secretion ability in the neurons,because of the readjustment of bioacitve molecules with LPS preconditioning in target cells.