Objective: Through observing the active influence of tongluojiunao(TLJN) injection on the normal and ischemic rat cerebral microvascular endothelial cells(CMECs) and primary discussion on total protein secretive time-effect feature in cellular conditioned medium,to establish a technical basis for further study of solubility protein.Methods: After TLJN injection having reacted upon normal and ischemic cerebral microvascular endothelial cells(CMECs),MTS/PMS colorimetric method,Bradford assay and colorimetric analysis antigenic were used respectively and isochronously to test cellular activity,total protein quantity of conditioned medium,and LDH releasing value of conditioned medium,observing them and analyzing the time-effect feature of cellular conditioned medium.Results: TLJN injection can not only promote the activity of ischemic cells,but also reduce LDH releasing value.In the normal group,the peak time of total protein quantity is at 3rdh,at this time,cell activity is highest and LDH releasing value is least.In the ischemia group,the peak time of total protein quantity is at 6thh,simultaneously, LDH releasing value is least,but cell activity is a little lower than that of 3rdh.Conclusion: TLJN injection has protective action on ischemic cells.In this study,it is suggested that regarding conditioned medium from 3rdh to 6thh as collecting aim is the best period to study biological effects of solubility protein.