Research On The Chemical Constituents And Processing Principles Of Himalayan Euphorbia Based On The Efficacy Of Mongolian Medicine

[Objective]To provide scientific basis for the clinical application of Euphorbia himalaye-nsis Boiss in Mongolian medicine,the efficacy and toxicity of Euphorbia himalayensis Boiss in Mongolian medicine was evaluated and the active fractions with the main efficacy of Mongolian medicine were screened out and the main chemical constituents and indicators of Euphorbia himalayensis Boiss were analyzed.By establishing the processing criterion of Euphorbia himalayensis Boiss and explaining the processing principle.the experimental data were provided for the clinical application and industrialization of Euphorbia himalayensis Boiss slices.By comparing the main phannacodynamic effects in before and after processing of Euphorbia himalayensis Boiss.this paper demonstrates the traditional efficacy characteristics of "if not processed.it can induce vomiting and if processed it has diarrheal effect "Mongolian medicine and the processing theory of "Change the efficacy of drugs to suit the condition"for the purpose of Mongolian medicine.[Methods]The diarrhea.emetic and diuresis effects and toxicity of Euphorbia himalayensis Boiss were evaluated by mouse diarrhea test(defecation time and filter paper trace method),rat emetic test(Heterophilia kaolin method),rat diuresis test(Metabolic cage method)and mouse acute toxicity test(improved Karber method):Euphorbia himalayensis Boiss was extracted from water.ethanol.petroleum ether.chloroform.ethyl acetate and n-butanol.used by decocting.reflux and extraction.The effective parts were selected by mouse diarrhea test(defecation time and filter paper trace method):UPLC-MS/MS was used to identification of the main chemical constituents of the purgative active fraction of Euphorbia himalayensis Boiss.The index components were screened by reference and plant properties of Medicinal Materials.A HPLC method for determination of Euphorbia himalayensis Boiss was established:Using the traditional appearance.the main pharmacodynamic effects and the total terpenoids content of the main active inigredients were observed as indicators.The optimum processing conditions were determined by orthogonal test of multi-index weighted comprehensive analysis.The quality standards of complying with the relevant provisions of the "Chinese Pharmacopoeia" and TLC identification method and HPLC content determination method were established:The methods of animal experiment which is including mouse diarrhea test,rat emetic test.acute toxicity test,rabbit skin irritation test.rabbit eye irritatioa test.mouse intestinal propulsion test and the analysis methods of UP-MS.BCA.PCA,OPLS-D and statistical methods was used to comared the purgative effect.emetic effects.toxicity.irritating and effects on intestinal motility.intestinal mucosa and intestinal tissue protein and variation of chemical composition in before and after processingof Euphorbia himalayensis Boiss.[Results](1)There were significant differences in time to start defecating and traces of loose stool within 6 hours of mice in each dosage groups of Euphorbia hinalayensis Boiss(P<0.01).There were significant differences in the amount of kaolin in the high dose group(P<0.01).There was no significant difference in low dose group of Ephorbia himalayensis Boiss(P>0.05).There was no significant difference in 6h urine volume among dosage groups(P>0.05).LDso by intragastric administration in mice was 9.63 ±1.54 g/kg.and the 950%confidence region was 8.2?11.29g/kg.(2)There were significant differences time to start defecating and traces of loose stool within 6 hours of mice in Euphorbia himalayensis Boiss alcohol extract group,petroleum ether fraction group and ethyl acetate fraction group(P<0.01).There was significant difference in chloroform fraction group(P<0.05).There was no significant difference in water extract group and n-butanol fraction group(P>0.05).(3)Twenty-two compounds were identified from the petroleum ether fraction of Euphorbia himalayensis Boiss,which is including three trterpenoids and 19 diteipenoids.Lanosterol had a good linear relationship in the range of 0.06mg/mL?0.16mg/mL(r=0.9999).the average recovery was 101.31%and the average content was 6.17mg/g.(4)The optimum processing condition of Euphorbia himalayensis Boiss are as follows:the proportion of medicinal materials and highland barley is 1:10,highland barley temperature is 120??roasting processing time is 40 minutes;Slices had clear TLC spots and good separation.Rf=0.39.It is stipulated that the moisture content should not exceed 8.0%and the total ash content should not exceed 7.0%and acid insoluble ash should not exceed 3.00%and alcohol soluble extract(hot leaching method)should not be less than 15.0%and lanosterol content should not be less than 3.Omg/g.(5)There were significant differences in tine to start defecating and traces of loose stool within 6 hours of mice in each dosage groups of processed Euphorbia himalayensis Boiss(P<0.01).There were no significant differences in the amount of kaolin in each dose group(P>0.05).LD50 by intragastric administration in mice was 14.04±1.92g/kg.and the 95%confidence region was 12.25?16.08g/kg.Compared with before processing.the cathartic effect was reduced by about 10%,the emetic effect almost disappeared,and the toxicity was reduced by about 45%.Processed Euphorbia himalayensis Boiss group has mild irritation to normal skin of rabbits.moderate irritation to rabbit eyes,and can obviously promote the promotion rate of activated carbon in small intestine of mice(P<O.Olor P<0.05).At the same time,only a small number of intestinal villus tip epithelial cells in the small intestine were stained lightly.Sixteen differentially expressed proteins were extracted and identified from the small intestine of mice in the raw and processed groups of Euphorbia himalayensis Boiss were closely related to 13 biological processes.5 molecular functions,1 cell location and thnee metabolic pathways,which are including Mucin type O-glycaii biosynthesis,Prion diseases and Base excision repairs.The mainly significant difference marker of Euphorbia himalayana Boiss before and after processing was Geraniin.8-O-Tigloylimgol,Helioscopinolide E.17-Hydroxyjolkinolide B and so on.[Conclusion]Euphorbia himalayensis Boiss has significant pharmacological activity in inducing diarrhea and emetic.but has not diuretic effect.It has certain toxicity and belongs to moderately toxic drug.Its petroleum ether fraction is the main active part of catharsis,which mainly contains diterpenes and triterpenes.The processing method of Highland Barley of Euphorbia himalayensis Boiss not only conforms to the traditional processing requirements of Mongolian medicine.but also conforms to the relevant requirements of "Chinese pharmacopoeia".Purgative effect of Euphorbia himalayensis Boiss was obviously alleviated after processing.Emetic effects becoming insignificant is associated with reduced strong irritating toxicity,which was also related to intensive stimulation promotes small intestinal propulsion and intestinal villus lesions of small intestinal wall,inflaminatoiy cell exudation from small intestinal mucosa and proteins in small intestinal tissues mainly composed of AnxalO.Dctppl and St6galnac.The material basis is terpenoids represented by 8-O-Tigloylingol.Helioscopinolide E and 17-Hydroxyjolkinolide B.