Effect Of Tripterygium Glucoside And Its Compatibility With Radices Paeoniae Alba On MC3T3-E1 Cell’s Proliferation、 Cycle、Apoptosis And Related Proteins

Objective:This study explores the influence of Triptolide、Tripterygium glucosides and its compatibility with Radices paeoniae alba on MC3T3-E1 cell’s proliferation、cycle、apoptosis and related proteins by using the cell biological methods, it is aim to provide experimental basis for the treatment of Tripterygium Preparation on ankylosing spondylitis(AS) and effect on its osteogenesis, it also provides some theoretical support for the clinical treatment of the compatibility of Tripterygium wilfordii.Methods:1.Culturing MC3T3-E1 cells in vitro, using BrdU、flow cytometry and hochest method to detect cell’s proliferation、cycle and apoptosis after the cells are handled by different doses of Triptolide; Besides, we also use western blot to detect the expression of the proliferation、cycle and apoptosis proteins (PCNA; Cyclin D1, Cyclin E; Bcl-2, Bax, cleaved-caspase-3).2.We set up Triptolide、Tripterygium glucosides、Tripterygium glucosides add Radices paeoniae alba、Radices paeoniae alba and blank control groups, MTT was performed to detect proliferation of each group; western blot was performed to detect the expression of cycle and apoptosis proteins(Cyclin Dl, Cyclin E; Bcl-2, Bax and cleaved-caspase-3). Results:1.Effect of different doses of Triptolide on MC3T3-E1 cell’s proliferation、cycle、 apoptosis and related proteins:(1)BrdU shows that compared with the OnM group, the absorbance value in 7nM and 14nM groups decrease obviously;(2)The result of flow cytometry indicates that the percentage of MC3T3-E1 cell is increased along with the doses of Triptolide;(3)After hochest dyeing, we find apoptosis cells, flow cytometry shows that the late apoptosis rate are increased along with the dose of Triptolide;(4)Western blot shows that compared with the OnM group, the CyclinDl and Cyclin E in 7nM and 14nM groups are significantly reduced, and the Bax and cleaved-caspase-3 are significantly increased; the PCNA、and Bcl-2 are also significantly reduced in the 14nM group;2. Effect of Tripterygium glucosides and its compatibility with Radices paeoniae alba on MC3T3-E1 cell’s proliferation and cycle、apoptosis related proteins:(1)MTT indicates that the absorbance value of Tripterygium glucosides add Radices paeoniae alba, Tripterygium glucosides and Radices paeoniae alba groups decrease not obviously, while Triptolide can significantly reduce the absorbance value;(2)Western blot shows that:①Compare with the blank group, Bcl-2、Bax and cleaved-caspase-3 in Tripterygium glucosides add Radices paeoniae alba group are increased; compare with the Tripterygium glucosides group, Bcl-2 is increased while cleaved-caspase-3 are significantly decreased, Bax has no obvious change; compare with the Triptolide group, Bcl-2 is increased while Bax and cleaved-caspase-3 are significantly decreased;② Compare with the blank group, Cyclin D1、Cyclin E and Bcl-2 are decreased, while Bax and cleaved-caspase-3 are significantly increased in Triptolide and Tripterygium glucosides groups; compare with the Triptolide group, Bcl-2 in Tripterygium glucosides group is increased while Bax is decreased, cleaved-caspase-3 changes not obviously;③Bcl-2 is increased lightly in Radices paeoniae alba group, while Bax and cleaved-caspase-3 change not obviously.Conclution:1.MC3T3-E1 can be blocked at G0-G1 period by Triptolide;2.Triptolide can inhibit cell’s proliferation and apoptosis effectively, with a dose related effect;3. Tripterygium glucosides and its compatibility with Radices paeoniae alba have some effect on cell’s apoptosis, but not obvious on cell’s proliferation and cycle;4. Triptolide has the best effect on cell’s proliferation、cycle and apoptosis, Tripterygium glucosides is the second, effect of Tripterygium glucosides add Radices paeoniae alba is not obvious, while Radices paeoniae alba has almost no effect on MC3T3-E1 cell.