| Kansui Radix(the roots of Euphorbia kansui T.N.Liou ex T.P.Wang),which was bitter,cold,toxic,one of the most commonly used Chinese medicine.Kansui is widely used in clinical treatment of ascites due to cirrhosis,nephritis edema,infiltration of pleurisy and ascites due to advanced schistosomiasis in China for centuries.Kansui is only for external use when it is raw since it’s thrill and toxicity,and must be processed if it is to be taken orally,in order to enture the security of the clinical application.The main content of this paper includes the following parts:Part one:Literature researchBy referring the relevant ancient and modern literature,acquired a more systematic summary of diterpenoids in Euphorbia L.,from the common traditional Chinese medicine of Euphorbia L.with diterpenoids,the main classifications of diterpenoids,and biological activity of diterpenoids,Literature studies have revealed the main chemical components of Euphorbia L.contain diterpenoids,triterpenes,sterols,etc.And these diterpenoids generally have obvious pharmacological action such as anti-cancer,antibacterial,anti-virus,And most of the diterpenes contain 10 to 12 ring,alcohol or cyclic ester bond,Therefore,the study of the structure and toxicity of these compounds can provide scientific basis for the clinical safety of this kind of composition and medicinal plants of Euphorbia L.Part two:By studying the toxicity of zebrafish embryos,the composition ofgroups B was determined to be the most toxic ingredients of allUsing the zebrafish embryo model to evaluate the toxicity of different toxic group in kansui and Kansui Radix stir-baked with vinegar,found that the composition group B had the highest toxicity,composition group C less than group B,the composition group A had lowest toxicity,and the composition group D showed no toxicity in a certain dose range.At the same time,the toxicity of each composition group in kansui was significantly higher than this in Kansui Radix stir-baked with vinegar.Stir-baking Kansui Radix with vinegar can significantly reduce its toxicity,which provided significant data for promoting safer and better use of this herb in clinic.Part three:Component separation of ethyl acetate part of Kansui RadixBy using different elution gradient elution parts are-inverting the preparatio n of high performance liquid chromatography(UPLC)and the preparation of th in layer separation methods,such as,from euphorbia kansui parts,a total of 20 compounds were isolated from ethyl acetate,use the means of identification,such as TLC、UV、NMR identified 15 of them.Five ingenane-type diterpenes were named as kansuiphorinC(KS-03),3-O-(2’E,4’Z-decadienoyl)-20-O-acetyl-ingenol(KS-04),3-O-(2 ’E,4 ’E-decadienoyl)-20-O-acetylingenol(KS-05),3-O-benzoyl-20-deoxyingenol(KS-06),5-O-benzoyl-20-deoxyingenol(KS-07);three jatr-ophanetype diterpene was named as kansuinin A(KS-10),kansuinin D(KS-11),kansuininC(KS-12);five triterpenes,euphol(KS-13),kansenone(KS-08),epi-kansenone(KS-09),eupha-8,23-diene-3β,11β-diol-7-one(KS-01),tirucalla-8,23-diene-3β,11β-diol-7-one(KS-02);other types such as β-sitosterol(KS-14),isoscopletin(KS-15);KS-01 is the first time from euphorbia kansui,KS-02 is a new chemi-cal compound from this genus,From the cell experiment,KS-01 and KS-02 have better inhibit-ion action on HCT-116、MKN-45,With the increase of the concentration,the cell proliferation inhibition rate increased,It has a certain am-ountpoison relations,thus,we infer that compound KS-01 and KS-02 may be the main material basis of anti-cancer and anti-tumor for kansui.Part four:Quantify analysis of toxic ingredient groups of Kansui Radix stir-baked with vinegar and ethyl acetate extractBy using liquid chromatography-mass spectrometry technology to determine the content of pnrificatus compounds which were separated from Kansui,the result showed that Kansui Radix stir-baked with vinegar,in ethyl acetate extract,in addition to the content of 5-O-benzoyl-20-deoxyingenol increased by about 13.25%,the rest of the diterpenoids and the triterpenes content were decreased.For example,the decreasing rate of ingenane type diterpenes were 14.33%,12.78%,18.06%and 7.55%;the decreasing rate of jatrophane type diterpenes were 19.41%,10.37%,16.13%,12.45%and 11.43%;the decreasing rate of triterpenes were 15.80%and 14.51%.In the composition group of A,in addition to the content of 5-O-benzoyl-20-deoxyingenol increased by about 21.97%,the others were reduced.And the decreasing rate of ingenane type diterpenes were 14.18%and 16.09%;In the composition group of B,in addition to the content of 5-O-benzoyl-20-deoxyingenol increased by about 8.01%,the others were reduced.And the decreasing rate of ingenane type diterpenes were 8.67%and 11.43%;In the composition group of C,the decreasing rate of jatrophane type diterpenes were 12.76%,10.34%,9.08%,8.99%and 22.19%;And in the composition group of D,it basically did not contain diterpenoids and triterpenes.Based on the analysis of the above compositional changes and the results of the second chapter,speculated that the decrease of the terpenoids in Kansui has a good correlation with reducing toxicity by the processing of the vinegar.because it has the same parent structure as the KPC,so that speculated the KPC may hydrolyze and translate into 5-O-benzoyl-20-deoxyingenol,so that it increases.Part five:Discussing the changes of diterpenes that stir-baked with vinegar by through comparison between chemical structure and toxicity of KS-04 and hydrolyzed productBecause of euphorbia kansui in the past dynasties vinegar main to reduce their toxicity,So the acid hydrolysis is adopted for KS-04 to discuss the chang es of diterpenes.First of all,we can acquire hydrolysis of KS-04 by acid hydrolysis;Secondly,The hydrolysate can be confirmed by TLC and UPLC;Thirdly,we can use the technology of Q-TOF to confirm the chemical structure of the hydrolysate;Finally,we use enterocyte IEC-6 to evaluate the toxicity of hydrolysate.Experimental results show that,after acid hydrolysis of the compound,a new substance can be prod-uced below the KS-04 of the thinlayer chromatography.And in the UPLC chromate-gram,got that the new material which is contented in ethyl acetate extract of Kansui Radix stir-baked with vinegar is higher,while the content in the corresponding raw product is lower.Analysing by the technology of Q-TOF,we find that this component-t may be ingenol,and its toxicity to IEC-6 cells was significantly less than KS-04.So that inferred the high toxicity compound of ingenol can be transformed into alcoh-ol with small toxicity.Maybe this is the mechanism of detoxification of Kansui Radix stir-baked with vinegar. |
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