目的:确定对鹿茸样品活性有保护作用的蛋白酶抑制剂种类及作用浓度。方法:选择金属蛋白酶抑制剂EDTA-2Na、胰蛋白酶抑制剂(Approtinin)和胃蛋白酶抑制剂(Pepstatin),以MTT法检测其对NRK-49F细胞增殖活性和对鹿茸促细胞增殖活性的影响以及对鹿茸样品33℃放置不同时间的保护作用。用Bradford法测鹿茸样品蛋白含量并调整到作用浓度400μg/mL。结果:当EDTA-2Na、Approtinin和Pepstatin浓度分别小于200、50、400μg/mL时对细胞增殖活性没有影响。EDTA-2Na浓度为6.25和12.5μg/mL时对鹿茸促增殖活性有加强作用,但对于鹿茸样品高温下放置没有明显的保护作用。12.5μg/mL的Approtinin对鹿茸促增殖活性有促进作用,并且对鹿茸样品高温放置有明显的保护作用,6.25μg/mL时效果最佳(P<0.05)。12.5μg/mL的Pepstatin表现出对鹿茸促增殖活性的加强作用,并表现出对鹿茸样品高温放置的保护作用,但数据无统计学差异。结论:3种蛋白酶抑制剂中胰蛋白酶抑制剂对鹿茸促细胞增殖活性有较好的保护作用。 Objective: To determine the type and concentration of protease inhibitors that have protective effects on the activity of antler samples. Methods: The metalloproteinase inhibitors EDTA-2Na, trypsin inhibitor (Approtinin) and pepsin inhibitor (Pepstatin) were selected and their effects on the proliferation of NRK-49F cells and the proliferation of antler cells were detected by MTT assay. Antler samples were placed 33 ℃ for different time protection. Determine the antler protein content with Bradford method and adjust to the concentration of 400μg / mL. Results: The cell proliferation activity was not affected when the concentrations of EDTA-2Na, Approtinin and Pepstatin were less than 200, 50 and 400μg / mL, respectively. EDTA-2Na concentration of 6.25 and 12.5μg / mL of velvet antler proliferation activity to enhance the role, but for the antler samples placed under high temperature no significant protective effect. Approtinin at 12.5μg / mL could promote the proliferation of pilose antlers and protect the pilose antler at high temperature, with the best effect at 6.25μg / mL (P <0.05). Pepstatin at 12.5 μg / mL showed enhanced antler proliferative activity and showed a protective effect on high-temperature placement of antler samples, but the data were not statistically different. Conclusion: The trypsin inhibitors of the three protease inhibitors have a good protective effect on the proliferation of antler cells.