尿多酸肽注射液体外生物学活性测定方法的建立

Development and Verification of a Novel in vitro Assay for Determination of Biological Activity of Uroacitides Injection

建立尿多酸肽注射液体外生物学活性检测方法,并对方法进行验证。采用磺酰罗丹明B(SRB)染色法,检测不同时间与不同浓度尿多酸肽注射液处理前后,人肝癌细胞HepG2增殖及凋亡情况。结果显示,尿多酸肽注射液对人肝癌细胞HepG2有明显的抑制作用,稀释度在1∶320～1∶2. 5范围内有很好的剂量反应关系,作用24,48,72,96 h后,随着药物浓度的增加及作用时间的延长,人肝癌细胞HepG2的存活率逐渐降低。辅料对人肝癌细胞HepG2无刺激作用,方法专属性符合要求,测定10批尿多酸肽注射液,结果显示该方法有很好的耐用性。尿多酸肽注射液在体外可抑制人肝癌细胞HepG2的增殖,可用于尿多酸肽注射液的体外生物学活性的评价和质量控制。

To establish a method for determination of the biological activity of uroacitides injection in vitro and to verify the method,uroacitides injection in vitro induced apoptosis of human liver tumor cell line HepG2,SRB staining and many methods were used to determine the effects of uroacitides injection on human liver tumor cell line HepG2,which could inhibit the proliferation and induce the apoptosis.Different concentrations of uroacitides injection had obvious inhibitory effect on the human liver tumor HepG2 cells,inhibitory effect was dose-and time-dependant within a certain range and the diluent had a good dose-response relationship in the range 1∶320~1∶2.5.After being inhibited for 24,48,72 and 96 h,with the increase of drug concentration and the extension of the action time,the cell survival rate was gradually decreased.Accumulation has no stimulative effect on human liver cancer cell line HepG2.The method specifically reviewed the requirements and determined 10 batches of uroacitides injection.The results show that the method has good durability.Uroacitides injection can inhibit the proliferation of human liver cancer cell HepG2 in vitro and can be used for the evaluation and quality control of in vitro biological activity of uroacitides injection.The new in vitro assay for uroacitides injection activity is developed and verified,which can be used for the biological activity of uroacitides injection.