Discrimination of proliferation inhibiting ingredients in Corydalis soxicola on rat hepatic stellate cell-T6 based on composition-activity relationship / 中草药

ABSTRACT

Objective: To establish the composition-activity relationship (CAR) model based on study of chemical composition and relating proliferation inhibitory rate of Corydalis soxicola and recognize the anti-hepatic fibrosis active compounds of C. soxicola. Methods: Nine orthogonal C. soxicola extract were analyzed by HPLC, and 21 characteristic peaks were profiled. Anti-hepatic fibrosis activity was investigated by MTT assays on HSC-T6, and the potential active components were identified by scores plot and variable importance in projection (VIP) values by means of orthogonal partial least squares (OPLS) analysis, and the activities of identified components were verified by MTT and flow cytometry. LDH kit was used to detect the effect of dehydrocavidine, palmatine, and berberine on LDH activity. Results: The results showed that six peaks including peaks 13, 14, 16, 18, 19, and 20 were significantly related to anti-hepatic fibrosis activity among nine orthogonal extract from C. soxicola. Peaks 18, 19, and 20 were characterized as dehydrocavidine, palmatine, and berberine, respectively. MTT assay showed that dehydrocavidine, palmatine, and berberine with various concentration significantly inhibited the proliferation of HSC-T6 cells. It was also found in flow cytometry that the apoptotic rates of dehydrocavidine, palmatine, and berberine (0.10 mg/mL) on HSC-T6 were 42.12%, 42.22%, and 36.73%, respectively, which were obviously higher than that of control (1.69%, P < 0.01). No obvious cytotoxic effect was found when the final concentration of dehydrocavidine, palmatine, and berberine were less than 0.15, 0.10, and 0.10 mg/mL, respectively. Conclusion: In this study, it was for the first time found that dehydrocavidine, palmatine, and berberine in C. soxicola extract can effectively inhibit the proliferation and induce apoptosis of HSC-T6 based on composition-activity relationship, and there was no obvious cytotoxic effect of the effective concentration, which showed that they may be the active components with potential anti-hepatic fibrosis effect and have a certain security in the application in C. soxicola. At the same time, it also suggests that the research ideas based on CAR can provide an effective method for the identification of active components in natural plants.