Preparation Of Durio Zibethinus Murr Shell Extract As Well As Its Anti-inflammatory And Analgetic Activities

BackgroundDurio zibethinus Murr, also named hainan rambutan fruit, is the durian fruit of the Bombacaceae, and is a kind of lardge tropical fresh fruits with unique flavor. Durian origined in India and Malaya, and then spreaded to the tropical countries of southern and southeast Asia such as Philippines, Tailand, Sri Lanka and Indonesia.Durian, which has a good reputation of "the king of tropical fruit" with food and medicine function, contains rich minerals and vitamins which are benefit for the health of human being. Recently, the researches of durian have mainly focused on genetics, breeding, nutritional and chemical ingredient, food processing as well as marketing. On the other hand, some modern reseaches demonstrated that there were plenty of amino acids and minerals in durian shell, such as Ca, K, Fe, which showed medicinal effects of durian shell. Some traditionary duian shell foods in southeast Asia and southern China, such as soup of duian shell, were used for "jianghuo and jiezhi", "buxue and yiqi", "zirun and yangyin". Howerer, the pharmacological activites of durian shell have been rarely reported. Accordingly, the studies ahout the chemical ingredient and the boiactivities of durian shell incur more and more attention in recent years.Objective(1) To compare the extraction methods of DSE, and study their optimum preparation.(2) To study the effects of DSE on anti-inflammatory, cough relieving, analgesia, antibiosis, which provide experimental evidence for the exploitation of DSE.Methods(1) DSE preparation tests:DSE was prepared by reflux extraction, microwave extraction, supersonic wave extraction and semi-bionic extraction. In order to assess the effects of all factors and acquire the optimum technology, orthogonal design was used for the experiment.(2) Anti-inflammation tests:50 male Kunming mice of 7 week-old were randomly divided into 5 groups, namely, normal control group (10 mg·kg-1 distilled water), positive control group (1 mg·kg-1 dexamethasone), DSE group(100 mg·kg-1 DSE), DSE group (300 mg·kg-1 DSE), DSE group (900 mg·kg-1 DSE). The anti-inflammation activities of DSE were examined by investigating its effects on peritonitis induced by acetic acid, on ear oedema of mice induced by croton oil, on paw edema of mice induced by carrageenin test, on ear-swelling and capillary permeability induced by 2,4-DNFB, and on hypodermic granulation proliferation induced by tampon.(3) Cough relieving tests:50 male Kunming mice of 7 week-old were randomly divided into 5 groups, namely, normal control group (10 mg·kg-1 distilled water), positive control group (20 mg·kg-1 codeine), DSE group(100 mg·kg-1 DSE), DSE group (300 mg·kg-1 DSE), DSE group (900 mg·kg-1 DSE). The effect of DSE on relieving cough was measured by cough models induced by ammonia and SO2 in mice. (4) Analgesia tests:50 male Kunming mice of 7 week-old were randomly divided into 5 groups, namely, normal control group (10 mg·kg-1 distilled water), positive control group (200 mg·kg-1 aspirin), DSE group (100 mg·kg-1 DSE), DSE group (300 mg·kg-1 DSE), DSE group (900 mg·kg-1 DSE). The effect of DSE on analgecize was examined by hot plate test and twisting reaction induced by acetic acid in mice.(5) Antibiosis tests:Constant dilution was used for detecting the DSE MIC of Staphylococcus aureus(ATCC 29253), Staphylococcus epidermidis(ATCC12228), Bacillus aeruginosus(ATCC 27853), E.coli (ATCC25922. And the disc-agar diffusion test was used for evaluating the effect of DSE on antibiosis.Results(1) The results of DSE preparation:As for the reflux extraction, the sequence of factors were ratio of solvent and material> reflux temperature> alcohol concentration> reflux time; The ratio of solvent and material could markedly affect the extraction (p<0.01). Reflux temperatue could significantly affect the extraction (p<0.05); The optimum technology was shown to require 30 min of reflux time,90℃of reflux temperature,80% alcohol as well as 50:1 ratio of solvent and material. As for the microwave extraction, the sequence of factors were microwave time> alcohol concentration> microwave power> ratio of solvent and material. Microwave time, alcohol concentration and microwave power could significantly affect the extraction (p<0.05); The optimum prepation required 20 min of microwave time, 800 W of microwave power,80% alcohol as well as 20:1 ratio of solvent and material. As for the supersonic wave extraction, the sequence of factors were supersonic wave power> supersonic wave time> ratio of solvent and material> alcohol concentration. Supersonic wave power could notablely affect the extraction (p< 0.01), supersonic wave time could also significantly affect the extraction (p< 0.05); The optimum technology required 20 min of supersonic wave time,500 W of supersonic wave power,20% alcohol as well as 20:1 ratio of solvent and material. As for the semi-bionic extraction, the sequence of factors were the first decoction pH>the second decoction pH>the third decoction pH> extraction time. The first decoction pH could significantly affect the extraction (p<0.05). The optimum technology required pH 4 at the first decoction, pH 5 at the the second decoction, pH 6 at the third decoction, and 30 min of extraction time.(2) The results of anti-inflammation tests:Compared with control group, all groups of DSE could effectively reduce the peritonitis induced by acetic acid in mice (p<0.001), decrease the croton oil-induced mice ear oedema and this effect of 300 and 900 mg·kg-1 DSE were significant (p< 0.05),100 and 300 mg·kg-1 DSE could markedly restrain the paw edema of mice induced by carrageenin (p<0.05),900 mg·kg-1 DSE could effectively inhibit the ear-swelling and capillary permeability induced by 2,4-DNFB in mice with good dose-effect relationship, and all groups could significantly repress the hypodermic granulation proliferation induced by tampon in mice with dose-dependent manner(p<0.001).(3) The result of cough relieving tests:Compared with control group,900 mg·kg-1 DSE could not only effectively prolong the latency but also obviously decrease the number of cough induced by ammonia and SO2 in mice(p< 0.05), and it was in dose-dependent manner.(4) The results of analgesia tests:Compared with control group,900 mg·kg-1 DSE could markedly extend the latency by acetic acid in mice(p<0.05), and significantly extend the analgesia time by hot plate test (p<0.05).(5) The results of antibiosis tests:Except for Bacillus aeruginosus, the effects of DSE on Staphylococcus aureus, Staphylococcus epidermidis, E. coli were very inferior. Conclusions(1) Compared with traditional reflux extraction, microwave extraction and supersonic wave extraction, especially semi-bionic extraction could prepare DSE efficiently, economically and environment friendly. Orthogonal design was not only a method to analyze the factors of DSE preparation, but also an easy method to acquire the optimum preparation of DSE.(2) 300 and 900 mg·kg-1 DSE had potential effects on some acute and chronic inflammations, and the mechanism might be partly related to its inhibitory effect on the metabolic product of arachidonic acid and its effect on enhancing the activities of immunological cell.(3) 900 mg·kg-1 DSE showed potential effects on relieving cough induced by chemical factors exciting pruritus of tunica mucosa bronchioru.(4) 900 mg·kg-1 DSE had potential protective effects on acute pain induced by chemical facor, and DSE exerted some effect on the pain induced by physical agent such as hyperthermia.(5) DSE showed extremely weak antibiosis activities for most bacteria, only some antibiosis activities for Bacillus aeruginosus.