Bovine Trabecular Meshwork Cells In Vitro High-pressure Culture Model Of The Effective Parts Of The Establishment And Traditional Chinese Medicine Drops Out Drops Ratio

Purpose: (1) To establish high pressure cell culture model of bovine trabecular meshwork(BTM) cells in vitro and observe effects of pressures on cultured BTM cells. (2)To approach the possibility of matching bolting on effective part's composition of Chinese herbs by cell line in vitro. (3) To study effects of the traditional Chinese medicine Di Ming humor ocular GD for BTM cells cultured in vitro.Methods: (1) Establish normal pressure cell culture model of BTM cells in vitro. (2)Set different pressure (20mmHg, 40mmHg, 60mmHg, 80mmHg) to BTM cells in tightly closed culture-bottle for 24 hours, then set the pressure of 80mmHg to BTM cells for 12, 24, or 48 hours, and collect BTM cells and measure the expression of actin and tubulin by immunohistochemical method. (3) Set BTM cells in the pressure of 80mmHg for 24 hours and observe the ultramicrostructure. (4) Put GD with different concentration and PBS into BTM cells cultured for 24 hour in vitro, then test the OD value of living cells by MTT after another 24 hours. (5) Add G4D6 into BTM cells, with one group in 80mmHg pressure, another in normal pressure, then measure the expression of actin and tubulin by immunohistochemical method after 24 hours. (6)Add G4D6 and latex beads into BTM cells, one group in 80mmHg pressure, another in normal pressure, then measure their phagotrophy ability after 24 hours.Results: (1)The pressure of 40mmHg, 60mmHg and 80mmHg for 24 hours can decrease the expression of actin and tubulin in BTM cells, and the pressure of 80mmHg for 24 or 48 hours can decrease the expression of actin, so does tubulin after 12,24,48 hours. (2)BTM cells' ultramicrostructure is damaged after 24 hours in 80mmHg. (3)With G8D2 0.00016mg/ml, 0.000032mg/ml etc, it can improve BTM cells proliferation. (4)With G4D6's concentration of 0.5mg/ml, living 24h in 80mmHg pressure, BTM cells' expression of actin and tubulin is improved. (5)G4D6 of 0.5mg/ml make no influence on BTM cells phagotrophy function.Conclusions: (1) BTM cells can survive under some pressure, and their expression of actin and tubulin can be decreased ,with direct correlation to increasement of pressure and duration time. (2) BTM cells obviously damaged after...