Study On The Anti-inflammatory And Analgesic Activity And Chemical Components Of The Ethyl Acetate Extract From Viburnum Taitoense Hayata

The Viburnum plants are more than 230 in the world,mainly distributed over the entire temperate area of the northern hemisphere,and a few extends to the tropics of South America and Southeast Asia.Viburnum plants have a medicinal history in my country,according to the research records in “Zhong Guo Zhi Wu Zhi” and "Zhong Guo Zhong Yao Zi Yuan Zhi Yao",there are about 74 species in my country,and are widely distributed in all provinces in the country.District,about 43 of them are used as Chinese herbal medicines.The roots,stems,leaves,and mature fruits of the Viburnum can be used in medicine,with heat-free,sputum dehumidification,spleen digestion,blood circulation and swelling,anti-phlegm and other effects.Viburnum taitoense Hayata,also known as Xuelizangzhu,Sijiqing(Guangxi),Lenghanmu(Xing’an)is a plant of the genus Viburnum,which is produced in northwest Guangxi,southern Hunan,and eastern Taiwan,and grows in stony shrubs or beside valleys and streams.The Guangxi Zhuang Autonomous Region is located in the southwest China,which is bordered in Vietnam.It is subtropical climate,local minority Zhuang,Miao,Yao people living in humid,cold mountains for a long time,so there is a wealth of understanding and practice for the treatment of fractures and rheumatoid diseases experience.In the mountainous area in northern Guangxi Zhuang Autonomous Region,Viburnum taitoense Hayata has exact efficacy due to its treatment of fell injuries.Therefore,local minorities often use the fresh branches of Viburnum taitoense Hayata with Sambucus williamsii Hance,used to treat fell injuries,fracture,eliminate wind and wetness.In addition,in the "Guang Xi Zhong Yao Zi Yuan Ming Lu",it records the branches of Taitong viburnum for colds,tatonymous diseases,and twisted,fractures."Zhong Guo Zhong Yao Zi Yuan Zhi Yao" also recorded the treatment of branches and leaves with Viburnum taitoense Hayata.Objective: In this paper,the anti-inflammatory and analgesic activities of ethyl acetate extract from Viburnum taitoense Hayata were determined by in vivo anti-inflammatory and analgesic experiments,and the chemical constituents of ethyl acetate extract from Viburnum taitoense Hayata were purified by silica gel column chromatography,gel chromatography and thin layer chromatography.It provides experimental basis for the application of Viburnum taitoense Hayata and the traditional efficacy of Viburnum taitoense Hayata.Methods: After the fresh branches and leaves of Viburnum taitoense Hayata were chopped,they were soaked and extracted 3 times with 3 times the amount of ethyl acetate at room temperature,each for 7 days.After filtration,the extracts were combined;the filter residue was soaked and extracted 3 times with 3 times the amount of 80% ethanol for 7 days each time,and concentrated under reduced pressure.The concentrated solution was added with water to obtain a suspension,and then extracted with ethyl acetate to obtain an ethyl acetate extract.The two ethyl acetate extracts were combined,concentrated under reduced pressure,and freeze-dried to obtain an ethyl acetate extract of Viburnum taitoense Hayata.Comprehensive use of modern chromatographic methods such as silica gel column chromatography,gel chromatography and thin-layer chromatography to systematically separate the chemical components of the ethyl acetate extract of Viburnum taitoense Hayata,and according to a series of modern spectroscopy methods such as proton nuclear magnetic resonance spectroscopy and carbon spectroscopy,and combined with the physical and chemical properties of the separated monomer compounds to identify their structure.The in vivo antiinflammatory and analgesic activity experiment of the ethyl acetate extract of Viburnum taitoense Hayata was determined by the inflammation and pain models of SD rats or KM mice.Inflammation model: carrageenan-induced rat foot swelling model,croton oil-induced mouse ear swelling model;pain model: acetic acid-induced mouse writhing model,formalin-induced pain model.Results: Using modern chromatographic techniques,a total of 9 compounds were separated from the ethyl acetate extract of Viburnum taitoense Hayata,which are(3S,5R,6R,8R,9S,10 R,14S,17S)-3β,6β-dihydroxy oleana-11,13(18)-dien-28-acid(1),ursolic acid(2),quercetin(3),daucosterol(4),β-sitosterol(5),lauric acid(6),Oleanol-18-ene-3-palmitate(7),3β-hydroxy-oleana-11,13(18)-dien-28-oic acid(8),oleanolic acid(9).In the anti-inflammatory experiment,the swelling degree of the rat’s foot after injection of carrageenan increased with time,and reached the maximum after 4 hours,while the volume decreased slightly after 5hours.Compared with the blank group,the middle-dose(70 mg/kg)and high-dose groups(140 mg/kg)of Viburnum taitoense Hayata ethyl acetate extract can be significantly improved in the first(0～2h)and second(after 2h)stages Inhibiting the swelling degree of rats induced by carrageenan(P < 0.01),showing a good anti-inflammatory effect,and its anti-inflammatory activity showed a significant concentration-dependent.In addition,it can be seen from the experimental results that the anti-inflammatory effect of the ethyl acetate extract of Viburnum taitoense Hayata reached the strongest at 4 h,and decreased at 5 h.Similarly,the results of the croton oil-induced ear swelling experiment showed that the high-dose ethyl acetate extract of Viburnum taitoense Hayata group(200 mg/kg)had a certain inhibitory effect on the ear swelling of mice(P<0.05).The effect also showed obvious concentration dependence.In addition,in analgesia experiments,Viburnum taitoense Hayata ethyl acetate extract showed significant analgesic effects on mouse pain models induced by acetic acid writhing and formalin-induced pain models(P < 0.01).Conclusion: Nine compounds were isolated from the ethyl acetate extract of Viburnum taitoense Hayata,and the activity of the isolated compounds was analyzed by consulting the literature,which indirectly clarified the antiinflammatory and analgesic activity of Taitung viburnum.In vivo experiments showed that the ethyl acetate extract of Viburnum taitoense Hayata had certain anti-inflammatory and analgesic effects,which provided experimental basis for the application of taitungensis and elucidation of its traditional efficacy.