Abstract
The herbal medicine Ipomoea stolonifera (I. stolonifera) has previously been shown to have considerable anti-inflammatory potential in vivo and in vitro. To establish a method for exploring the synergistic effects of multiple compounds, we study the compatibility and dose optimization of compounds isolated from n-butanol extract of I. stolonifera (BE-IS). Raw264.7 cell was treated with lipopolysaccharide (LPS) in the presence of compounds from BE-IS, namely scopoletin, umbelliferone, esculetin, hesperetin and curcumin, using the orthogonal design, uniform design and median-effect method. To verify the best efficacy of principal constituents in vivo, the uniform design was used in the croton oil-induced mouse ear edema model. The results from LPS-induced the production of prostaglandin E2 (PGE2) show that, esculetin, curcumin and hesperetin were the principal constituents that had synergistic effects when used at the optimal ratio. Additionally, the principal constituents were found to work synergistically in the croton oil-induced mouse ear edema model at low doses. It turned out that the three experimental optimization and analysis methods (orthogonal design, uniform design and median-effect method) can be effectively used to solve both compatibility and dose optimization for combined use of multiple compounds.
Highlights
To enhance therapeutic efficacy and reduce adverse effects, practitioners of traditional Chinese medicine (TCM) prescribe a combination of plant species/minerals, called formulae, based on clinical experience
Agents that are effective at controlling these pro-inflammatory mediators are considered to be promising therapeutics for these diseases[14], such as steroidal anti-inflammatory drugs (SAIDs) and nonsteroidal anti-inflammatory drugs (NSAIDs)
We first identified the optimal concentrations of principal constituents by uniform design, and using the median-effect method, we found that synergistic effects clearly exist between the principal constituents when they are used at the optimal concentrations
Summary
To enhance therapeutic efficacy and reduce adverse effects, practitioners of traditional Chinese medicine (TCM) prescribe a combination of plant species/minerals, called formulae, based on clinical experience. It remains a puzzle as to how to construct and identify a prescription containing the best combination and optimal doses to combat diseases To solve this question, we designed a series of methods to rapidly identify the compatibility and optimized doses of drugs in combination, and validated the methods using compounds isolated from n-butanol extract of Ipomoea stolonifera (BE-IS). Macrophages directly recognize pathogens, ingest them by phagocytosis and produce various pro-inflammatory mediators, including TNF-α,IL-1β,IL-6, NO and PGE2. These mediators play an essential role in strengthening the innate immune response, and in activating other immune cells to promote a secondary immune response[10]. The optimized combination of the bioactive compounds with fewer adverse effects could be developed for long-term administration
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