Influence Of Methylprednisolone In Combination With Mycophenolic Acid On The Proliferation Of Human Mesangial Cells

Objective1. To observe the biological characteristics of culcured human mesangial cells (HMCs),and master the basic process of cell culture.2. To investigate the effect of mycophenolic acid, methylprednisolone, and both in combination on proliferation, apoptosis and cell cycle of human mesangial cells.Clarify the relationships among drug dose, action time and effect.MethodsRecover the frozen mesangial cells in 37℃water bath and move it into 50 ml culture bottles,then it shuld be placed in 37℃,5% CO2 incubator. When adherent cells up to 80%, passage the cells and continue to culture, spread 3-4 generations, the cell viability was enough strong for the experiments.Mesangial cells were inoculated on culture plates. The first step is:Human mesangial cells were added different concentrations of mycophenolic acid and methylprednisolone,measured proliferation inhibition of HMC with MTT at 24h,48h and 72h at the 490nm wavelength.Found the proper dose of both,then detected the combined effects of proliferation inhibition after 48 h.The second step is:Human mesangial cells were added different concentrations of mycophenolic acid and methylprednisolone,measured cell cycle of mesangial cells with FCM after 48 h,then pick the groups of MPA 2.5μmol·L-1 and MP 5 mg·L-1, to detect the combined effects of both. The third step is:Human mesangial cells were added different concentrations of mycophenolic acid and methylprednisolone, measured apoptosis rate of mesangial cells with FCM after 48 h.Results1. Mycophenolic acid, methylprednisolone, and the combined effects of both concentration-dependently inhibited the proliferation of mesangial cells.When the concentrations of Mycophenolic acid more than 1μmol·L-1, the inhibition reached the maximum.The inhibition of Mycophenolic acid and Methylprednisolone reach the strongest role after 48h.These two drugs have a synergistic effect,the combined anti-proliferative effect of two drugs is superior to the effect of any drugs alone.2. Mycophenolic acid can arrest mesangial cell cycle in S phase, methylprednisolone may arrest mesangial cell cycle in G0/G1 phase, the combined effects of both can arrest in G0/G1 phase and S phase. Compared with the control group,the difference is statistically significant(p<0.05).3. Mycophenolic acid can promote the apoptosis of mesangial cells, while the low-dose methylprednisolone in mesangial cells have no effect on apoptosis.Conclusions1. Mycophenolic acid and Methylprednisolone significantly inhibited the proliferation of the cultured human mesangial cells, the combined anti-proliferative effect of two drugs is superior to the effect of any drugs alone.2. The inhibitive effect of mycophenolic acid and Methylprednisolone on proliferation of mesangial cells probably is related to the promoting apoptosis of cells and orderly blocking the cell cycle.3. This study from the perspective of inhibiting cell cycle to investigate mycophenolic acid and methylprednisolone on the proliferation inhibition of cultured human mesangial cell, especially provides a theoretical basis for the sequential combination therapy of cell cycle.