The Research On Radix Glycyrrhizae Reducing The Toxicity Of Radix Aconiti Carmichaeli

The combinatiing of Radix Glycyrrhizae and Radix Aconite Lateralis Preparata has the effect of increasing efficacy and decreasing toxicity. This study started from the phenomena of ingredients self-assembly in the mixed fried decoction of Radix Glycyrrhizae with Radix Aconite Lateralis Preparata. The research objects were the protein of Radix Glycyrrhizae and aconitine (AC) which was the major toxic component of Radix Aconite Lateralis Preparata. Explored the possibility of the establishment of the hypotheisi "the toxicity reduced because of hazardous substances were presented in micelles". This study included the experiments of Radix Glycyrrhizae decreasing the toxicicty of Radix Aconite Lateralis Preparata, optimization the conditions of purification and self-assembled of natural protein in Radix Glycyrrhizae, drug embedded, the studing on toxicity and stability of the composite nanoparticles.HPLC analysis showed that the content of AC is 16.14 μg/mL in Radix Glycyrrhiza decoction, the content of AC was 3.11 μg/mL in the mixed fried decoction of Radix Glycyrrhizae with Radix Aconite Lateralis Preparata, however the content of AC was 96.66 μg/g in Radix Glycyrrhiza. The result of intraperitoneal injection experiment showed that the mice were poisoned by Radix Glycyrrhiza decoction e, but not by the mixed fried decoction of Radix Glycyrrhizae with Radix Aconite Lateralis Preparata. Combineing with toxicity test and the content changes of toxic substances verified that Radix Glycyrrhiza can reduce the toxic of Radix Aconite Lateralis Preparata.A new protein GP has been separated and isolated from licorice by High Q anionic exchange and POROS@R1 hydrophobic interaction chromatography, buffer extraction and ethanol sedimentation before. This protein showed a single band in SDS-PAGE, which molecular weight was 31.0 kDa, and the concentration of it was 0.4564 mg/mL.The concentration 0.4564 mg/mL of GP could form stable particles only at pH5.7 in normal temperature, particle mean diameter is 206.20±2.02 nm, it illustrated that GP has the characteristics of self-assembling. The 0.4564 mg/mL of GP with 10 μg AC (HPLC grade methanol solution) could form the GP-AC particles which particle mean diameter was 238.20±1.23 nm, and Zeta potential was 10.50±0.62 mV,and light scattered count value is 302 kcps at pH 5.0 in normal temperature. Under the same conditions, just joining same volume of methanol,the particle mean diameter of GP-AC particles was 183.50.20±123 nm.Characterization of GP-AC particles, found that its particle mean diameter increases by 30.0%.than it under the same conditions (uncontain AC) formed by the particle size GP-AC particles, and the mass rate of the AC and GP was about 9.2:1, GP for AC embedding rate reached 28.22%.Intraperitoneal injection of 50.0 ug/mL AC standard solution and GP-AC particle suspension, found that the AC standard solution can poisone mice, GP-AC particles suspension do not. It showed that GP is one of the attenuated composition, and it can reduce toxicity by embedding AC.Analyzing different influence to the stability of the GP-AC particles, and the results showed that it can be stored in normal temperature for short-term.Combined with animal toxicity experiments and content changes of the toxic substances, and then built the GP-AC particles model by self-assembly methods, proved Radix Glycyrrhiza can reduce the toxic of Radix Aconite Lateralis Preparata., and try to explain the mechanism of toxicity reduction.