Study Of The Constituents And Toxicity Changes With The Combination Of Veratrum Nigrum And Ginseng In Vitro And In Vivo

Objective: Shi Ba Fan is an important part of compatibility theory of traditionalChinese medicines (TCM), and plays important roles in safe and reasonable use ofTCM. The compatibility of ginseng and veratrum nigrum is one of the typical pairs inTCM. Formerly studies were mainly focused on identification the phenomenon ofincreased toxicity in the condition of combination, but the mechanism is still unclear,especially the toxic constitutes change and its relation with toxicity in vivo and in vitro.Therefore, the aim of the current study was to explore what constitutes changed, andwhether the toxicity was also changed together. The results showed some evidence ofconstitutes change with toxicity change. Above evidences contributed to understandthe regular of the eighteen incompatible pairs more deeply and provided moreinformation on scientific and resonable use of TCM.Method: With the platform of UPLC/TOF-MS and method of acute toxicity of mice,the current study initially observed chemical fingerprint o f veratrum and inentified thepeaks of toxic constitutes. The changes of the chemical component of differentmatchedbetween ginseng and veratrum nigrum in vivo and in votro were alsocompared.The toxicity evaluating systerm of veratrum and ginseng was founded. TheLD50and MTD of veratrum and ginseng were assayed respectively, and the differencebetween co-decoction and combination of veratrum and ginseng. The evaluation ofacute toxicity by two defferent model-homogeneous design and different percentage of ginseng were accomplised, compared the merits and demerits of each method. In theresearch of homogeneous design of acute toxicity, combination of ginseng andveratrum caued increaed toxicity at some specified proportionality, more researchwere done according this clue. In vivo studies, the sub-enzyme of CYP450whichcontribute for the metabolism of alkloids of veratrum nigrum were chiefly cleared.Subsequently the levels of main alkloids of vetrum nigrum in blood of betweenginseng treated group and blank group were determinded, some evidences of arise oftoxicity of ginseng and veratrum nigrum in the enzyme of CYP450level were found.Result: According to the acute toxicity of each herb in mice, the LD50of veratrumnigrum in mice of oral drug administration was2.566g/kg. And the MTD of ginsengwas over80g/kg. when matched veratrum nigrum and ginseng with1:1, the death rateof mixed and co-decoction was35%and50%respective, and it has no principle ofstatistics obviously. The result of homogeneous design indicated that the use ofveratrum nigrum and ginseng with1:2.63, the death rate of mixed and co-decoctionwas10%and70%respective, ewhich has principle of statistics. It was showed thatwhen veratrum nigrum and ginseng was matched with1:2.63, Toxicity alkaloid seriessuch as jervine, veratrosine,3-angeloylzygadenine was changed in different ways.From different dosage (the use of veratrum nigrum and ginseng was10:1to1:10), itdisplayed that the death rate of mice was decreased as the rise of compatibility ofginseng. From different dosage experiment, we found that keep the veratrum nigrumunchanded, as the rise of compatibility of ginseng, toxic alkaloid series such asveratrosine, Veratrum alkamine,3-angeloylzygadenine, germidine, veratramine,Rubijervine were decreasrded regularly. Derivative of jervine was also decreaseded butveratrosine was up rise when extended the time of decoction. Through the research of influence of inhibitor of CYP450on the metabolism ofalkaloids of veratrum nigrum.Extracting alkaloids from veratrum nigrum firstly, then,assay the content of jervine and veratramine was21.82%and24.60%respectively.Research on5sub-enzyme of CYP450, it showed that the addition of inhibitor ofCYP1A2can inhibit metabolism of diverse alkaloids significant, while the inhibitor ofCYP2B1, CYP2D1, CYP2E1, CYP3A4cannot inhibitor the metabolism of alkaloidsof veratrum nigrum notable.After oral drug administration ginseng7days9multiple maximum dose level of clinicalcontinuously, then peritoneal injection alkaloids once on the eighth day. Observe thedifferent between blank and ginseng group. It showed that the AUC of jervine andveratramine of ginseng group was higher than blank..Conclusion: As the change tendency of veratramine, jervine, veratrosine, Veratrumalkamine, germidine were same to the change tendency of death rate of mice, thesealkaloids maybe contribute to the toxic effect of ginseng and veratrum nigrum.CYP1A2was the main enzyme responsible for the metabolism of diverse alkaloids.Ginseng may change the activity of sub-enzyme of CYP450of rats, resulted slowingdown the metabolism of alkaloids of veratrum nigrum.