ABSTRACT
Objective: To research the process of extraction and purification of water-soluble alkaloids of Aconiti Lateralis Radix Praeparata (ALRP). Methods: Orthogonal design L9(34) and single factor experiments were employed to optimize the extraction conditions using UPLC-MS/MS. The total extraction yield of 14 alkaloids (karacoline, songorine, fuziline, neoline, talatisamine, mesaconine, aconine, hypaconine, benzoylmeaconine, benzoylaconine, benzoylhypaconine, mesaconitine, aconitine, and hypaconitine) was used as an index. The absorption-desorption characteristics of five macroporous resins were evaluated to optimize purification conditions using contents of eight water-soluble alkaloids (aconine, mesaconine, hypaconine, fuziline, neoline, talatisamine, songorine, and karacoline) as indexes. Results: The optimized extraction and purification conditions were as follows: steamed tablets were decocted thrice with 10-fold pH 3.5 acidic water for 2 h each time (maintaining pH 3.5). The alkalinity of water decoction was adjusted to pH 10.0 with 20% NaOH and then heated for 2 h. The supernatant was adjusted to pH 11.0 by 20% NaOH and then was subjected to macroporous resin HPD300 whose relative adsorption amount was 2.5 g dried medicinal herb/mL resin, and then the resin was eluted with 6 BV of water and 4 BV of 80% ethanol respectively. The 80% ethanol fraction was evaporated under vacuum to give the water-soluble alkaloids extract. The extract yield was 1.69%. The total contents of mesaconine, aconine, hypaconine, fuziline, neoline, talatisamine, karacoline and songorine were above 15%. Conclusion: The optimized extraction and purification process is stable and feasible, and this present study provides the reference for the research and application of water-soluble alkaloids in ALRP.
ABSTRACT
Objective To observe the anti-inflammatory effects of three kinds of aconine from Radix Aconiti Lateralis Preparata ( Fuzi) on macrophages, as well as the effects of their combination with sinomenine. Methods The effect of three monoester alkaloids from Fuzi, benzoylaconine ( BAC) , benzoylmesaconine ( BMA) and benzoylhypaconine ( BHA) , on RAW264.7 cells proliferation was detected by cell counting kit 8 (CCK8) in vitro. RAW 264.7 macrophage cells were stimulated by lipopolysaccharides (LPS) and then treated with different concentrations of BAC, BMA, and BHA. Tumor necrosis factor alpha ( TNF-α) and interleukin 6 (IL-6) secreted by RAW264.7 cells were measured by enzyme-linked immunosorbent assay (ELISA) . The anti-inflammatory effects of three monoester alkaloids combined with sinomenine were evaluated by Zhengjun Jin Q method. Results The concentrations of TNF-α and IL-6 secreted by LPS-stimulated RAW264.7 cells were increased significantly ( P<0.01) , and then were inhibited by BAC ( 20, 30 μmol/L) , BMA ( 40, 80, 160μmol/L) and BHA ( 40, 80, 160 μmol/L) significantly ( P<0.01) . Antagonistic effects were present when 30 μmol/L of BAC or 160 μmol/L of BMA or 160 μmol/L of BHA was respectively used together with 100 or 300 μmol/L of sinomenine (Q<0.85). Conclusion The three kinds of monoester alkaloids from Fuzi exert anti-inflammatory effect on macrophages, and the effective dose of BAC is lower than that of BMA and BHA. The combination of BAC, BMA or BHA at the analyzed dosage with sinomenine has antagonistic effect.
ABSTRACT
Objective: To investigate the regulation of in vitro myocardial cell L type calcium channel of rats with active component of Shenfu Decoction and its combination. Methods: With Langendorff cardiac infusion, the single ventricular myocardial cell with acute enzymylosis approach was acquired and the voltage current of L type calcium channel was recorded with whole cell patch technique. Results: The active ginsenosides Re (3, 10, 30, and 100 μmol/L), and Rg1 (10 and 100 μmol/L) can reversibly reduce L type calcium current in a dose dependant manner, Rb1 (10 and 100 μmol/L) has no effect on L type calcium current; the active component aconine (10 and 100 μmol/L) of aconite could irreversibly enhance L type calcium current in a dose dependant manner; The combination of ginsenosides Re (20 μmol/L) and Rg1 (80 μmol/L) also reduced the L type calcium current, which is more significant than either ginsenosides Rg1 and Re; The inhibitory rate of the combination of ginsenosides Rg1, Re and aconine (100 μmol/L) has no difference from the combination of ginsenosides Rg1 and Re. Conclusion: The active component ginsenosides Rg1 and Re of Shenfu Decoction can reduce the L type calcium current, while aconine can irreversibly enhance the L type calcium current; The combination of ginsenosides Rg1 and Re can obviously increase the reduction of L type calcium current, while the combination of ginsenosides Rg1, Re and anonine can not change the reduction of L type calcium current.