The Effect Of Hypertonic Glucose And Intermittent High Glucose Of MMP-9 Producted By Mononuclear Macrophages

ObjectiveTo observe the effect of glucose of different concentrations and its fluctuation on MMP-9 expression in mononuclear macrophages and to study whether or not osmotic pressure changes participate in the process in which intermittenthighglucose affects MMP-9 expressed by mononuclear macrophages, mononuclear macrophages cultured in vitro induced from THP-1 cell. The aim is to study the importance of intermittent high glucose in the mechanism of diabetic marcovascular disease, so as to provide the oretical supports and exp-erimen tal bases for the treatment of. diabetic marcovascular disease.Methods1. THP-1 cell culture:taken out from the fridge of-80℃, the THP-1 cell strains were thawed on a superclean bench and then grafted to a culture flask which was put in an incubator with 5% CO2 (37℃).the cultured monocytes would be stained by Trypan blue to calculate the survival percentage.2. mononuclear macrophages obtaining:incubated with PMA for 48 hours, the THP-1 cells induced into its mononuclear macrophages.3. mononuclear macrophages identifying:incubated with the RBC of chicken to identify the phagocytosis of mononuclear macrophages.4. Grouping:according to glucose concentration and its fluctuation, mononuclear macrophages were divided into 4 groups:contrast(5 mmol/L), hypertonic glucose(15 mmol/L), intermittent high glucose (5-15 mmol/L),and osmotic pressure controlled (mannitol 5-15 mmol/L)5. Test of MMP-9:RT-PCR was used to test MMP-9 expression in mononuclear macrophages,comparing the differences for statistical significance.Results 1. MMP-9 expression in hypertonic glucose,intermittent high glucose and osmotic pressure controlled are much higher than those in the contrast(P<0.01).2. MMP-9 expression in intermittent high glucose are much higher than those in hypertonic glucose (P<0.01)3. MMP-9 expression in osmotic pressure controlled are between those in hypertonic glucose and intermittent high glucose, but compared with them, it is statistically more significant (P<0.01)Conclusion1. Glucose of higher concentration can lead to a marked increasement of mononuclear macrophages to express more MMP-9.2. Glucose of intermittent high glucose enhances mononuclear macrophages express more MMP-9 than glucose of high concentration.3. The concentration fluctuation of glucose in promoting mononuclear macrophages to express MMP-9 is partly related with osmotic pressure change.