Abstract:

BACKGROUND: Total soluble protein of Zaocys Dhumnades should be extracted and analyzed for searching active component. However, methods for extracting total protein from Zaocys Dhumnades are few.  
OBJECTIVE: To establish the method for extracting total soluble protein from Zaocys Dhumnades and observing its in vitro cytotoxicity.
METHODS: Schizolysis enzyme combined with salting-out method were used to extract the total soluble protein of Zaocys Dhumnades, and then the protein level was measured by Bradford and separated by SDS-PAGE. The band sizes and their differences were analyzed with quantity one 4.4. The liquid for extracting protein and different concentrations (0.5, 5, 50, 150, 450 and 900 mg/L) of total soluble protein of Zaocys Dhumnades purified by (NH4)2SO4, were used to culture fibroblast-like synoviocytes. The cell proliferation was analyzed by MTT.
RESULTS AND CONCLUSION: There was no significant differences between extraction amount of total soluble protein without and with being purified by (NH4)2SO4 (P > 0.05). The liquid for extracting protein and total soluble protein of Zaocys Dhumnades in concentrations of 150, 450 mg/L significantly inhibited fibroblast-like synoviocytes proliferation; while total soluble protein of Zaocys Dhumnades in concentrations of 0.5, 5 and 50 mg/L had no effect on the fibroblast-like synoviocytes proliferation in the control group. The findings demonstrated that, proteins of Zaocys Dhumnade can be extracted effectively with low cytotoxicity using schizolysis enzyme combined with salting-out method, which suitable for in vitro cell study.