Bioactive consistency evaluation of cardiotonic pill using marker genes / 药学学报

ABSTRACT

The study is aimed to develop a method in evaluation of the bioactive consistency of cardiotonic pill (CP). HepG2 cell line was employed as a biological detector. After treated with CP for 24 h, gene chip and qRT-PCR were used to select mRNAs that can represent the bioactivity of CP. Then similarity between different batches of CP were calculated based on expression levels of marker genes to evaluate the bioactive consistency of CP. Marker genes were selected according to the criteria as follows:① fold change 1.5; ② potential relevance to curative effects; ③ extensive involvement in the cellular functions and clustering analysis categories; ④ dose-dependent effect. A total of 10 genes were selected as bioactive markers of CP. Angular cosine was calculated to evaluate the similarity between two samples. The method was validated using intra-day precision and inter-day precision. Using angular cosine similarity, the intra-day and inter-day precision were 0.4% and 0.6%, respectively. The similarities of 6 batches of CDPs ranged from 0.992 to 0.999, and 1 batch of Compound Danshen Tablet was 0.534. The established method is specific and accurate, and provides comprehensive and objective evaluation of bioactive quality of CDPs. It can also benefit the bioactive consistency evaluation of other compounds in traditional Chinese medicines.