Studies On Different Compatibilities Of Veratrilla Baillonii Franch And Aconitum Brachypodum Diels

OBJECTIVE:To analyse the synergistic mechanism of co-decoction of Aconitum brachypodum Die1?ABD?and Veratrilla baillonii Franch?VBF?,and mixed decoction of chloroform fration of Aconitum brachypodum Diels?CFA?and water extract of Veratrilla baillonii Franch?WVBF?,thus compare the difference between two extraction processes and discuss the toxicity change before and after compatibility through mice experiment,analgesic and anti-inflammation effects of mixed decoction and co-decoction through rats experiment before and after compatibility.Also,to analyse substance changes of compatibility and find the best compatibility.METHODS:?1?The changes of chemical compositions between mixed decoction and co-decoction were analysed and the mechanism was discussed using HPLC and NMR methods.?2?The heart rate was calculated and morphologic changes were observed in the cardiovascular system of 10 zebra fish selected at random under a stereo microscope.The optimum concentration was determined and 4different compatibility proportions of CFA and WVBF were set to do the same experiment under the concentration to evaluate the cardiac toxicity using zebra fish mode.?3?Up-and-down method and improved Karber test was used to calculate the median lethal dose(LD₅₀)of mixed decoction and co-decoction of different compatibility proportion to compare toxicity before and after compatibility.?4?Acetic acid induced writhing response and hot plate methods were used to observe and compare central analgesia and peripheral analgesic aspects of CFA group,1:2 and 1:5group of mixed decoction;Xylene-induced KM mice ear edema and hot water-induced tail-flick tests were utilized to evaluate the anti-inflammatory and analgesic effect of ABD decoction and 3 different compatibility proportions?1:1,1:2,1:5?of co-decoction.RESULTS:?1?The HPLC analysis suggested the contents of mixed decoction before and after compatibility didn't have obvious changes compared with ABD decoction.However,the peaks of co-decoction were obviously different compared with CFA.There are 4 obvious new chromatographic peaks in compatibility group,and the structures were the compositions of VBF decoction identified by NMR.?2?In the cardiac toxicity evaluation experiment,other concentration groups didn't have effect on heart rate except 200 mg·L^-1group,and didn't induce arrhythmia of zebra fish.The concentration groups of 66.6 and 200mg·L^-1can induce toxicity phenotype.In different compatibility of CFA and WVBF experiment,experimental groups induce obvious cardiac toxicity except 1:5 group.?3?The result of p-and-down test suggested the LD50 of compatibility of CFA and WVBF?5:1,2:1,1:2,1:5?increasing respectively,all are higher than the LD50 of CFA.Improved Karber test result indicated that the LD₅₀ of 5 compatibilities?5:1,2:1,1:1,1:2,1:5?were between that of ABD and VBF,which indicated the acute toxicity was reduced through the compatibility.?4?The result of central analgesia experiment indicated that the 1:2 group and 1:5 group can enhance foot pain threshold,the analgesia effects of 1:2 group(CFA:WVBF,20 mg·kg^-1)as well as CFA(20 mg·kg^-1)group(200 mg·kg^-1)are significant,which were similar to that of aspirin group(200mg·kg^-1),and the analgesia percentages were similar to CFA group and slightly higher than aspirin group after 90 minutes,which indicated 1:2 group could reduce toxicity and keep efficacy.The analgesia percentage of 1:5 group(20 mg·kg^-1)gradually increased after 30,60,90 minutes,and obviously higher than that of CFA(20 mg·kg^-1)group,which indicated 1:5 group could reduce toxicity and enhance efficacy and extend efficacy.?5?The result of writhing experiment suggested that two groups of different proportions of CFA and WVBF groups could inhibit acetic acid induced writhing response.The writhing inhibition rates of 1:2 groups(20 mg·kg^-1)were lower than that of CFA group(20 mg·kg^-1).The writhing inhibition rate of the1:5group(20 mg·kg^-1)was slightly higher than that of CFA group(20 mg·kg^-1),which indicates the 1:2 group(20 mg·kg^-1)could reduce toxicity and decrease efficacy but the 1:5 group(20 mg·kg^-1)could slightly reduce toxicity and enhance analgesic efficacy in analgesic aspects.?6?The Xylene-induced ear edema test revealed that compared to control group,all the 3 different proportion groups of co-decoction between ABD and VBF groups could inhibit Xylene induced ear edema response,The edema inhibition rates of the 1:1(125 mg·kg^-1)as well as 1:2(875 mg·kg^-1)groups were lower than that of ABD decoction(125 mg·kg^-1)and Indomethacin group(20mg·kg^-1).The edema inhibition rate of the 1:5 group(1750 mg·kg^-1)was similar to Indomethacin group but slightly lower than that of ABD decoction group(125mg·kg^-1),which indicates that 1:1 and 1:2 group could reduce toxicity and decrease efficacy but the 1:5 group could reduce toxicity and keep anti-inflammatory efficacy.?7?In the hot water-induced tail-flick test,ABD decoction group(125 mg·kg^-1)and1:1,1:2 as well as 1:5 group of co-decoction all could enhance tail-immersion response threshold compared with control group.The analgesia percentage of ABD decoction group(125 mg·kg^-1)is the highest among all the groups,and the analgesia percentage of 1:1(500 mg·kg^-1)and 1:2 group(875 mg·kg^-1)were similar,The analgesia effects are inferior to Indomethacin group and the analgesia percentage of1:5 group(1750 mg·kg^-1)was slightly higher than Indomethacin group.The analgesia percentage of all the co-decoction groups in the 90 min were lower than that in the 30min,which suggested the analgesia effect is time-dependengtly decreasing.Compared to ABD decoction group,all the co-decoction groups could reduce toxicity and reduce efficacy.CONCLUSIONS:The compatibility of the co-decotion between VBF and ABD can increase the dissolution of some xanthones of VBF.Both mixed decotion and co-decotion of different compatibilities could decrease thetoxicity of CFA,and the detoxification efficacy increased along with the increasing proportion of VBF.1:5group,showed best effect of anti-inflammatory and analgesia effects,which provides scientific basis for clinical medicine rationally.