| Geopropolis is a high viscosity solid formed by stingless bees(Apidae,Trigona)collecting different plant shoots,resin secretions,and sand or soil in nature,and then mixing with its own secretions.Geopropolis has a unique and complex chemical composition affected by the stingless bee species,the geographical environment,local plant glue source difference and the different time of plastic extraction,etc.Geopropolis is mainly composed with flavonoids,phenolic acids,terpenoids,plant sterols,xanthones,vitamins,minerals,free fatty acids,hydrolyzable tannins,saponins,alkaloids,etc.Geopropolis has a variety of biological activities which were related to complex chemical composition of stingless bee propolis,such as anti-oxidation,bacteriostatic,anti-tumor,anti-proliferative,anti-inflammatory,anti-viral,anti-mutation,enhance immunity,etc.Different from the propolis collected by the Apis mellifera bees,the high content of terpenoids in the geopropolis is an important material basis for its biological activity and a major source of volatile components,however,current related research of geopropolis is insufficientHeterotrigona itama(H.itama)is a stingless bee species widely distributed in the tropics,this experiment used H.itama propolis collected from Sarawak,Malaysia as the research object,and studied the optimal extraction and related activities of terpenoids in H.itama propolis.The main research results are as follows:1.The single-factor experimental design and response surface were used to optimize the extraction of terpenoids in H.itama propolis.The best process parameters were obtained:95%ethanol extracted H.itama propolis for 72 h,material to liquid ratio 1:30 g/mL,when leaching 3 times,the terpenoids in the H.itama propolis had the maximum extraction rate(46.44±0.07%)2.Analyze the relative content of major terpenoids in H.itama propolis by LC-MS/MS(Liquid Chromatography-Secondary Mass Spectrometry)according to the five existing standards and preparation of pure products in the laboratory.The results showed that the H.itama propolis contains(24E)-Dammara-20,24-dien-26-al 0.926 mg/g,a-amyrin 1.029 mg/g,24(E)-cycloart-24-ene-26-ol-3-one 0.304 mg/g,20-hydroxy-24-dammaren-3-one 0.343 mg/g,borneol ketone 0.441 mg/g.3.The antioxidant and anti-inflammatory activities the stingless bee H.itama propolis were evaluated by vitro scavenging free radical experiment and the bacterial lipopolysaccharide(LPS)induced mouse macrophage inflammation experimental model.In vitro results showed that the DPPH free radicals and ABTS free radical scavenging IC50 of H.itama propolis extractions were 630.31±0.76 ?g/ml and 321.58±3.67 ?g/ml,respectively.For cell experimental,it is indicated that the optimized extractions of H.itama propolis can inhibit the relative expression of proinflammatory gene iNOS(Inducible Nitric Oxide Synthase),IL-1?(Interleukin-1?),IL-10(Interleukin-10)mRNA and the promote the relative expression of antioxidant gene HO-1(Heme Oxygenase 1)to inhibit the inflammatory responseIn summary,this subject optimized the extraction process parameters of terpenoids in H.itama propolis;Quantified five major terpenoids in the optimized extraction of H.itama propolis;It was verified that terpenoids in H.itama propolis has good anti-inflammatory activity and certain antioxidant activity.The results of this study can be used as a theoretical basis to promote the development and production of H.itama propolis resources. |
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