The Dsx And Their Constituents On Cultured Rat Retinal Ganglion Cells In Experimental Research

PURPOSE: 1 To build high pressure cell culture model in vitro of Retinal Ganglion Cells(RGCs) and observe the RGCs effected by pressure . 2 To research the sequence of screening of Chinese medicine component with cell line in vitro. 3 To study the effect and mechanism of DSX and DSX1-7 for RGCs cultured in vitro.METHODS: 1 Using one to three day SD-rats, we build Rat Retinal cells and RGCs cultured in virto.2 Put the different pressure (20mmHg, 40mmHg, 60mmHg, 80mmHg) to RGCs in close tightly culture-bottle for one to four hours, then collect RGCs and check the sum of apoptosis cells by Flow Cytometry(FCM). 3 Put different concentration DSX and DSX1-7, Nerve growth factor(NGF) and Boric acid Buffer in Retinal Nerve Cells (RNCs) and RGCs cultured after 24 hour in virto, then test the OD of living cells by MTT after anther 24 hours. (4) After cultured with the pressure of 80mmHg four hours, RGCs were collected and check the sum of apoptosis cells by Flow Cytometry.RESULTS: 1 RNCs can survive two or three weeks, RGCs can survive two or three days. 2 The pressure of 20mmHg, 40mmHg, 60mmHg and 80mmHg for one or four hours can acceleration the apoptosis of RGCs, but the apoptosis in the pressure of SOmmHg for four hours was the most severe in all. 3 (1) To RNCs , The ODs of living cells in NGF and DSX whose concentration was 5mg/ml , 0.5mg/ml, 0.05mg/ml and 0.005mg/ml are higher than control's (P<0.05). (2) To RNCs, The OD of living cells in DSX3 whose concentration was 10mg/ml is lower than control's (P<0.01). The ODs of living cells in NGF and DSX3 whose concentration was 2mg/ml, Img/ml, 0.2mg/ml and 0.1mg/ml, 0.02mg/ml and 0.01mg/ml is higher than control's (P<0.05~0.01). (3). To RNCs, The ODs ofliving cells in NGF and DSX7 whose concentration was 2mg/ml, Img/ml, 0.2mg/ml and O.lmg/ml are higher than contrors(P<0.05~0.01) .(4) To RGCs, The OD of living cells in DSX3 whose concentration was 2mg/ml and Img/ml are higher than control's(P<0.05) . The ODs of living cells in DSX7 which concentration was 2mg/ml, Img/ml and 0.2mg/ml are higher than control's (P<0.05~0.01).4 To RGCs with 80mg/ml for 4 hours , the percent of apoptosis of RGCs in DSX7 which concentration was Img/ml is lower than control's and model's (P<0.01).CONCLUSIONS: 1 The surviving time of RNCs was longer than that of RGCs.2 Pressure can accelerate the apoptosis of RGCs. The more high pressure and the more long time, the sum of apoptosis of RGCs is more.3 DSX7 was the most effictive factor in Protecting RGCs in vitro.4 It was a effective and cheap method using cells cultured in vitro to search effective factor in Chinese medicine.