Seasonal Variation In The Chemical Compositions Of Mytilus Crassitesta Lischlk And The Study Of Its Mechanism Of Anti-inflammatory Effect

Mytilus sp. is famous for its rich nutrients which are benefit for humans, and peopleoften called it as'marine egg'. Mytilus sp. is rich in protein, high composition of PUFA, andhas various minerals and 8 essential amino acids. Freeze-dried New Zealand green-lippedmussel powder (Perna canaliculus) and its stabilized lipid extract, Lyprinol, haveanti-inflammatory effect without any side effect including the damage to gastrointestinaltract caused by traditional anti-inflammatory drugs-NSAIDs. And Mytilus crassitestaLischlk produced from Shengsi in Zhejiang province have been shown that it contains richnutrients and the most significant finding is that it has the same anti-inflammatory activitywith Lyprinol. With Mytilus crassitesta Lischlk from four seasons, we have compared somenutrients of Mytilus crassitesta Lischlk from four seasons including fatty acids, amino acids,minerals and other common nutrients to evaluate which season has the highest nutritionalvalue; and with Mytilus crassitesta Lischlk from summer, we have studied how to producesoft capsule with the anti-inflammatory activity; and we also have compared five kinds ofoil with the lipid extract from the spring Mytilus crassitesta Lischlk in the aspect ofanti-inflammatory activity to explore the mechanism of the activity from lipid extract of thespring Mytilus crassitesta Lischlk. Some of the major conclusions are as follows:1. With GC for fatty acids determination, HPLC for amino acids and ICP-MS forminerals, we carried out a comprehensive evaluation of the nutritional values between fourseasons' Mytilus crassitesta Lischlk. The summer Mytilus crassitesta Lischlk had the highestamount in PUFA, n-3 PUFA and EPA+DHA; and the essential amino acids of the autumnMytilus crassitesta Lischlk are higher than other three seasons, and the essential amino acidscompositions from four seasons fulfill the necessities for all population (except 2-monthbaby) to grow.2. In establishing the procedure of anti-inflammatory soft capsule from supercriticalfluid (CO₂) extraction of freeze-dried stabilized Mytilus crassitesta Lischlk mussel powder,we found that the best condition to extract the lipid after the mono-and multiplicity analysisis as follows: extraction temperature is 45℃, the flow of CO₂ is 10kg/h, and the extractionpressure is 30MPa, on which conditions the extraction yield is 5.1ï¼…. With the mixture of extraction and soybean oil (1:1, V/V), we completed the soft capsule containing 200mgmixture in soft capsule machine.3. During the establishment of the procedure of soft capsule, we carried out anotherstudy on the stability in fatty acids of freeze-dried stabilized Mytilus crassitesta Lischlkmussel powder and its lipid extraction, and the influence of different extraction processes onthe fatty acids profile of lipid extraction. In the study of the stability of mussel powder, wecompared the fatty acids of the powder under two different temperatures (-18℃and 4℃) onthe 0day, 30day, 60day and 120day. We found that the fatty acids including C18:2n-6,C20:2n-6, C20:3n-6, C20:4n-6, C18:3n-3, C20:3n-3 are more steady than that stored on 4℃conditions. With the same study method, we found that the lipid extraction under 4℃fullwith N₂ is the most steady in fatty acids composition during storage which is also the mosteconomical, and we also found that the lipid extraction from supercritical fluid (CO₂)extraction of freeze-dried stabilized Mytilus crassitesta Lischlk mussel powder is verySteady for it contains much CO₂ in it after extraction and we suggest to use some measuresto protect the oil oxidation from light which is most major factor promoting fatty acidsoxidation. After comparing the influence of different extraction methods includingsupersonic extraction, supercritical fluid (CO₂) extraction, C:M (chloroform-methanol)extraction and Soxhlet Extraction, we found that the lipid from C:M (chloroform-methanol)extraction and Soxhlet Extraction had more n-3 PUFA (1603.40 and 1641.54 mg/100grespectively) than other two extraction methods, but in consideration of the security ofextraction on humans, we suggest to produce lipid extraction with supercritical fluid (CO₂)extraction without any solvent residual.4. In cotton pellet-induced granuloma model, 7 groups (8 rats per group) includingLyprinol group, the lipid extraction of the spring Mytilus crassitesta Lischlk group,profound fish oil group, Thunnus thynnus oil group, oleum lini group and Indometacin groupwere compared in their anti-inflammatory activity which lasted for one week. Theconclusion is that the inhibition effect by the lipid extraction of the spring Mytihtscrassitesta Lischlk and Lyprinol is the highest (37.48ï¼…and 36.99ï¼…respectively), and theinhibitory activity by other three oils are more lower (the highest is only 20.13ï¼…byThunnus thynnus oil). Further analysis of five kinds of oil showed that the amount of PUFA in the lipid extraction of the spring Mytilus crassitesta Lischlk and Lyprinol is no more thanthat in other three oils, and the compositions of EPA+DHA, n-3PUFA, n-6PUFA, MUFAand PUFA from Lyprinol are nearly the same as that in profound fish oil, and the lipidextraction of the spring Mytilus crassitesta Lischlk is even less than two fish oils. So wemay conclude that the strong anti-inflammatory activity which the lipid extraction of thespring Mytilus crassitesta Lischlk and Lyprino owned have nothing to do with thecomposition of n-3PUFA, n-6PUFA, MUFA and PUFA. And then we further explored thefatty acids profiles in livers and blood serum from the rats after experiment above. Theresults showed that there is no any significant difference in PUFA (especially in n-3 PUFA)composition between groups revealed that the PUFA in the lipid extraction of the springMytilus crassitesta Lischlk can not offer the anti-inflammatory activity or play a major rolein anti-inflammatory effect, and they may contain a group of lipid soluble active compoundswhich give the lipid extraction strong anti-inflammatory activity.