[Bioactive consistency evaluation of cardiotonic pill using marker genes]

Area of Science:

• Pharmacology
• Molecular Biology
• Traditional Chinese Medicine

Background:

• Ensuring the bioactive consistency of traditional Chinese medicines (TCMs) like cardiotonic pills (CP) is crucial for therapeutic efficacy.
• Existing methods may not fully capture the complex bioactivity of TCMs.
• HepG2 cell line offers a sensitive biological system for assessing cellular responses to pharmaceutical compounds.

Purpose of the Study:

• To develop and validate a novel method for evaluating the bioactive consistency of cardiotonic pills (CP).
• To identify reliable mRNA markers that reflect the bioactivity of CP.
• To establish a quantitative measure for comparing the bioactivity across different batches of CP.

Main Methods:

• Utilized HepG2 cells as a biological detector to assess CP effects.
• Employed gene chip and quantitative reverse transcription polymerase chain reaction (qRT-PCR) to identify 10 key mRNA markers.
• Selected marker genes based on fold change, relevance to curative effects, cellular function involvement, and dose-dependent effects.
• Calculated similarity between samples using angular cosine based on marker gene expression levels.

Main Results:

• Identified 10 specific mRNA markers representing the bioactivity of CP.
• Angular cosine similarity demonstrated high intra-day (0.4%) and inter-day (0.6%) precision.
• Six batches of CP showed high similarity (0.992–0.999), while one batch of Compound Danshen Tablet had lower similarity (0.534).

Conclusions:

• The developed method provides a specific, accurate, comprehensive, and objective evaluation of CP bioactive quality.
• This approach is valuable for ensuring the bioactive consistency of cardiotonic pills.
• The methodology can be extended to assess the bioactive consistency of other TCM compounds.