| Radix Aconiti is the main root of Aconitum carmichaeli Debx whichbelongs to the family of Ranunculaceae. Aconitum is mainly used forwind-cold-wetness bype of arthralgia，cold wet-arthralgia, joint pain, coldhernial pain and so on. It has an excellent analgesic effect. It mainly containsaconitine, mesaconitine，Hypaconitine and Benzoyl compounds of thesematerials which are the major toxic compositions because they all haveacetylation in their hydroxyl of8th and have aromatization in their hydroxylof14th. The toxic components are also effective components of analgesiceffect of Aconitum carmichaeli Debx. Pinellia ternata is the tuber of thePinelli a ternata(Thunb)Breit which belongs to the family of Araceae. Pinelliaternata have the effects of drying dampness and resolving phlegm, dissolvinglumps and resolving masses, and even can be used to relieve swelling and painwhen it is used externally.Aconite combined with pinellia ternate was one of the medical pairs ofeighteen incompatibilities, which belongs to the scope of taboos of traditionalChinese medicine, opposite to two drugs with severe side effects. Whetheropposing herbal remedies can use, talk to doctors through the age’scommunity. Some doctors believe that opposing herbal remedies increases thetoxicity, damage the body, thus stressed that they cannot be used. And thereare many opposing herbal remedies with documented thought they can play acomplementary and resistance claimed the product’s effectiveness. Modernclinical and experimental study on the same, shared or not controversies, andliterature, clinical observations and experimental research is currently nouniform set of conclusions, descriptions of their research is to make long andhard, deep and careful work. This study aims to examine effects of different proportions of raw pinellia ternate on toxicity, efficacy, and the dissolution-outamount of alkaloid of raw radix aconite.Part1Effect of Raw Pinellia Ternata on Acute Toxicity of Raw RadixAconiteObjective:To observe the effects of raw pinellia ternata in toxicity to raw radixaconiti, and the relation to the proportion of raw pinellia ternata, by using theLD50in mice as indicator, and to provide experimental basis for clinicalmedication.Method:210ICR mice of either sex,10were used as the NC group, the rest200mice were randomly divided into4groups in which Raw Radix Aconiticombined with different proportions of Raw Pinellia Ternata (Raw RadixAconiti: Raw Pinellia Ternata were1:0,1:0.25,1:0.5,1:1), with50mice ineach group and they were divided into5groups with different dosages. Thedrugs were ig administered to mice only once. The main toxic reaction (eg.cough, dyspnea, convulsion, drooling, loose stools) and death of mice wereobserved for14days, and95%confidence limit of the LD50were calculated.Result:The LD50and95%confidence limit of raw Radix Aconiti and rawPinellia ternata with different proportion respectively were:6.214g/kg(4.254-9.078g/kg)in group1:0;9.065g/kg(6.849-11.998g/kg)in group1:0.25;9.472g/kg(6.648-13.497g/kg)in group1:0.5;11.825g/kg(9.089-15.387g/kg)in group1:1. It showed that no significant difference in the incubation periodand duration of toxic reaction was observed between the groups. LD50increased with proportion of raw Pinellia ternata increasing.Conclusion:Raw pinellia ternata can significantly reduce the toxicity of raw radixaconiti. It has dose-dependent relation with the compatibility proportion of raw pinellia ternata.Part2Effect of Raw Pinellia Ternata on Analgesic Effect of Raw RadixAconitiObjective:1To observe effect of raw pinellia ternata on analgesic effect of rawradix aconiti and the relation of their compatibility proportion, and thusdiscuss the effect of raw pinellia ternata on analgesic activity to raw radixaconite.2To compare the difference of analgesic activity between soaking liquidsand water decoction of raw pinellia ternata and raw radix aconite combined inthe same proportion, and discuss the effect of decoction on analgesic activityof raw radix aconite.Methods:The hot plate test and classical writhing test were used to evaluate theeffect of the decoction and raw pinellia ternate in analgesic effect and therelation relation to the compatibility proportion of raw pinellia ternata.1The analgesic effect of the soaking liquids:50ICR mice were dividedinto NC group and raw radix aconite combined with raw pinellia ternate in1:0,1:0.25,1:0.5,1:1randomly, the dose of raw radix aconite was0.311g/kg ineach group,(concentration of raw radix aconite:15.55mg/kg), the volume was20ml/kg, the same volume of distilled water was gived to the NC group.2The analgesic effect of the water decoction: the compatibilityproportion was the same as in1.150ICR mice were divided into3teams inwhich the dose of the raw radix aconite were0.311g/kg,1.556g/kg,3.110g/kg(the concentration of raw radix aconite were15.55mg/ml,77.75mg/ml,155.50mg/ml),50mice in each team, and the mice were further divided intoNC group and the different compatibility proportions groups which raw radixaconite combined with raw pinellia ternata.Results:In the test of55℃hot plate,the dose of soaking liquids of raw radix aconite was0.311g/kg, the pain thresholds of groups(1:0,1:0.25,1:0.5,1:1)were all significantly higher than the NC group,(P＜0.05or P＜0.01),compared with the group of1:0, the pain thresholds of groups (1:0.25,1:0.5,1:1) were all reduced significantly,(P＜0.05or P＜0.01).When the dose of water decoction of raw radix aconite was0.311g/kg,the pain thresholds of groups (1:0,1:0.25,1:0.5) were all significantly higherthan the NC group,(P＜0.05or P＜0.01), compared with the group of1:0, thepain thresholds of groups (1:0.25,1:0.5,1:1) were all reduced significantly,(P＜0.05or P＜0.01).When the dose of water decoction of raw radix aconite was1.556g/kgand3.110g/kg, the pain thresholds of groups (1:0,1:0.25,1:0.5,1:1) were allsignificantly higher than the NC group,(P＜0.05or P＜0.01). compared withthe group of1:0, the pain thresholds of groups (1:0.25,1:0.5,1:1) were allreduced significantly,(P＜0.05or P＜0.01).In the test of writhing by acetic acid, the writhing time of the soakingliquids groups (1:0,1:0.25,1:0.5,1:1) were all significantly lower than the NCgroup,(P＜0.05or P＜0.01), compared with the group of1:0, the writhingtime of groups (1:0.25,1:0.5,1:1) were all increased significantly,(P＜0.05orP＜0.01).When the dose of water decoction of raw radix aconite was0.311g/kg,1.556g/kg, and3.110g/kg, the writhing time of the water decoction groups(1:0,1:0.25,1:0.5) were all significantly lower than the NC group,(P＜0.05orP＜0.01), compared with the group of1:0, the writhing time of groups (1:0.25,1:0.5,1:1) were all increased significantly,(P＜0.05or P＜0.01).Conclusion1Raw pinellia ternate can obviously reduce the analgesic effect of rawradix aconite, and there was significantly dose-dependent relationship to theraw pinellia ternate.2The analgesic effect of water decoction was weaker than the soakingliquids in the case of the compatibility proportion of raw pinellia ternate andthe dose of rew radix aconite was equal. 3Heating decoction can significantly reduce the analgesic activity of rawradix aconite.Part3Effect of Raw Pinellia Ternate on Dissolution-out Amount ofAlkaloid of Raw Radix AconitiObjective:To determine the content of aconitine, mesaconitine and hypaconitine insoaking liquid and water decoction of raw radix aconiti which combined withdifferent proportion of raw pinellia ternata, to observe the effect of decoctionand raw pinellia ternata on dissolving-out amount of alkaloid of raw radixaconite, to compare the difference between the soaking liquids and waterdecoction when the proportion of raw pinellia ternate was equal. And todiscuss the relation between the raw pinellia ternata in reducing toxicity andefficacy of raw radix aconiti and dissolving-out amount of the component oftoxicity and efficacy.Methods:1The preparation of soaking liquids: The proportion of raw radix aconitiand raw pinellia ternata was1:0,1:0.125,1:0.25,1:0.5,1:0.75,1:1and1:2,respectively. Mixture sample was soaked with water (1:8) for3times, each for24h. Extracts were combined and extracted triple with an equal volume ofdiethyl ether for3times. The diethyl ether extracts were combined andsubsequent evaporation of the solvent at20℃. The extraction solution wasobtained by adding dichloromethane.2The preparation of water decoction: The proportion of raw radix aconitiand raw pinellia ternata was1:0,1:0.125,1:0.25,1:0.5,1:0.75,1:1and1:2,respectively. Mixture sample was soaked with water (1:8) for2hours, boiledthem out for3times, the first for1hour, second for30minutes, the third for15minutes. Extracts were combined and extracted triple with an equal volumeof diethyl ether for3times. The diethyl ether extracts were combined andsubsequent evaporation of the solvent at20℃. The extraction solution wasobtained by adding dichloromethane. 3Determining the content of the alkaloid: The separation was performedon a Hypersil BDS C18column(4.6×250mm,5μm) and methanol-water-chloroform-triethylamine (70:30:1:0.2) were used as mobile phase with aflow rate of1.0ml/min. UV detection was set at235nm and columntemperature was30℃.Results:The standard calibration curves of aconitine， mesaconitine andhypaconitine were liner in the rang of1.5～25.0μg/ml(r=0.9996,17.0～85.0μg/ml(r=0.9998),16.0～160.0μg/ml(r=0.9996), respectively. This methodwas applied to simultaneous determination of three aconitine alkaloids in thegroups which raw radix aconiti: raw pinellia ternata were1:0,1:0.125,1:0.25,1:0.5,1:0.75,1:1,1:2, respectively. The contents of aconitine, mesaconitineand hypaconitine in each group were0.02164mg/g,0.08447mg/g,0.12517mg/g in1:0,0.01645mg/g,0.06822mg/g,0.10975mg/g in1:0.125,0.01369mg/g,0.05569mg/g,0.09710mg/g in1:0.25,0.01005mg/g,0.04424mg/g,0.06306mg/g in1:0.5,0.00618mg/g,0.03291mg/g,0.034798mg/g in1:0.75,0.01228mg/g,0.05193mg/g,0.10321mg/g in1:1.0.00359mg/g,0.01221mg/g,0.01837mg/g in1:2. There was no aconitine,mesaconitine, and hypaconitine in the water decoction in each group whichraw radix aconite combined with different proportion of raw pinellia ternate.Conclusion:1Raw pinellia ternata can significantly reduce the dissolving-out amountof alkaloid in raw radix aconiti, and the dissolving-out amount of alkaloid wasnegatively related with the compatibility proportion of raw pinellia ternata. Itshows that raw pinellia ternata reduces the dissolving-out amount of alkaloidin raw radix aconite may be one of the factors which raw pinellia ternatareduces the toxicity and efficacy of raw radix aconiti.2Decoction can significantly reduce the dissolving-out amount ofalkaloid in raw radix aconite. |
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