ABSTRACT
In order to research the mechanism of guiding action of borneol in Suxiaojiuxin pills, the model of in vitro intestinal absorption, in vivo drug metabolism of mice and cell in vitro absorption model of Caco-2 were established firstly. All animal experiments were in accordance with the regulations of the Animal Ethics Committee of Nankai University. The results showed that the cumulative absorption quantity and absorption permeability of ferulic acid and ligustilide in the intestinal juice of Suxiaojiuxin pills group were significantly increased comparing with fake Suxiaojiuxin pills group, which don't contain borneol. By using borneol, the content of ferulic acid and ligustilide in the blood and tissues, such as heart, were added. The transepithelial resistance value and the content of horseradish peroxidase (HRP) in Caco-2 were rapidly decreased and increased, respectively. Due to further explore mechanism of promoting intestinal absorption of borneol for drugs, in this study, photosensitive probes of borneol were synthesized to capture its targets, and dual luciferase reporter system was used to evaluate its activity of calcium. It was found that it could make calcium overload by regulating transient receptor potential cation channel, subfamily M, member 8 (TrpM8). Then, the results of mass spectrometry imaging showed that the accumulation of ferulic acid in the heart was significantly increased by borneol, and the relaxation rate of rat thoracic aorta was enhanced obviously. In summary, the borneol in Suxiaojiuxin pills can expand cell space and increase intestinal permeability by acting on TrpM8, thus promoting the intestinal absorption, tissue distribution and target organ enrichment of drugs.
ABSTRACT
Objective: By screening the quality markers of Ca2+ antagonistic ingredients, a rapid evaluation system about the vasodilatory effect of Angelica sinensis (AS) by near infrared spectroscopy (NIR) was established. Methods: The Ca2+ antagonists in AS were screened by UPLC/Q-TOF combined with Ca2+ double luciferase reporter gene system. To establish the quality markers, the antagonistic effects were further evaluated in cells and in vitro. The quality markers in multi batches of AS were quantitatively analyzed, and the corresponding NIR was obtained, and then the NIR fitting algorithm was established. Meanwhile, the relationship between Ca2+ antagonistic holistic activity of AS extract and the content of quality markers was investigated, and the prediction model for vasodilation efficacy of AS was constructed based on quality markers check analysis via NIR technique. Results: The screening result showed that ligustilide (X1) and levistilide A (X2) in AS had significant Ca2+ antagonistic effects, and the change of the content was consistent with the capacity of Ca2+ antagonistic action of AS extracts, so they were confirmed as quality markers. According to nonlinear regression analysis, the relationship between Ca2+ antagonistic effect (Y) of AS and the content of two quality markers satisfied the following functions: Y = 31.257 9 X1 + 381.352 0 X2 - 248.979 0 X1X2 + 18.482 2. In addition, the measured values from NIR simulation method for ligustilide and levistilide A detection showed a good correlation with the predicted values. Conclusion: In this study, it was found that ligustilide and levistilide A were quality markers responsible for vasodilation efficacy in AS, and the quantitative correlation between quality markers and the function of vasodilation effect was established. With the help of NIR technology, a novel solution for the rapid monitoring of the quality of traditional Chinese medicine was demonstrated.