Biological Efficacy Of Se-enriched Ligustrum Lucidum Fruit

Selenium (Se) element is one of the essential trace elements for bodies, which have some important biological functions, such as the antioxidation, immuno-enhancement activity on bodies and so on. In this study, we sprayed the sodium selenite solutions to the leaves of Ligustrum Lucidum of full-bloom stage to obtain the se-enriched Ligustrum Lucidum Fruit (Se-LLF) and study the biology activities of Se-LLF. The main contents include:the production of Se-LLF, detection of the nutritional and biological-active components, the security test of Se-LLF, the effects of Se-LLF on the growth of hybridoma cells and the immune function of animals.Test1Production of Se-LLF and determination of its mineral elementsIn the full-bloom stage of Ligustrum Lucidum, we prayed sodium selenite solutions with the concentration of1000,500,100mg/L in June2009, the same amount of tap water was sprayed in the negative control group,6trees in each group, a blooming spray was conducted a week later. Se-LLF were abtained until the fruits growed purple plump with natural growth of the privet trees in December2009. After the pre-treatment of Se-LLF,14mineral elements were determined by ICP-MS or FAAS. The results were listed as follows:phenomenons of blossom dropping and yellow leaves were observed while the leaves of the privet trees were treated with selenite sodium at the dose of1000mg/L, Once the concentration was500mg/L or100mg/L, the selenium contents of Se-LLF were up to0.93,0.43μg/g respectively, which showed an significant increase compared with the control group content with a concentration of0.03μg/g (P＜0.01), the contents of heavy metals including Pb and Cd were consistent with the national safety standards.Test2Effects of selenium on the natritional and active components of LLFThe effects of selenium on the nutritional components and bioactive components accumulation of LLF were observed through assaying of nutritional components, mineral elements and main active component such as oleanolic acid, ursolic acid of Se-LLF. The results showed that:protein, ursolic acid and amino acids in LLF were increased by spraying the sodium selenite solutions to the leaves of of Ligustrum Lucidum, and contents of many minerals were also remarkly increased, for instance, some essential elements such as selenium, strontium and manganese, while the contents of other constant elements such as sodium was significantly reduced. The oleanolic acid of Se-LLF was lower than that of the control group; the ursolic acid of Se-LLF increased with a certain positive correlation.Test3The security test of Se-LLFTo observe the acute toxicity of the Se-LLF, the maximum administration dose of the water solution was given to the mice in a day. The results demonstrated that the maximum dose of Se-LLF was80g/kg. Then the subchronic toxicity test was conducted with the rats by successive intragastric administration everyday during90days with the Se-LLF solutions at the dose of5000,2500and1250mg/kg respectively. The clinical symptoms were observed everyday during the experiment and the blood physiology, serum biochemical indexes and the changs of the tissues incuding heart, liver, spleen, lung, kidney and gastrointestinal were observed at the45days and90days. The results showed that the physiology and biochemical indexes of the experimental groups were within the normal range, there was no significant difference compared to the control group, which indicate that Se-LLF has neither acute toxicity nor subchronic toxicity under the experimental conditions. Test4The effect of Se-LLF on hybridoma cells proliferationTo determine the safety concentrations of aqueous extract and ethanol extract of Se-LLF and LLF for the growth of tumor cells by MTT method, and to analyze the effects of the drugs mentioned above on hybridoma cells proliferation under the safety concentration after incubated for12,24,36,48h respectively. The result showed that:Se-LLF and LLF aqueous extract and alcohol extract had no obvious toxicity on tumor cells at the dose of200,400μg/mL respectively. Se-LLF aqueous extract demonstrated promotive effects on hybridoma cells proliferation at various concentrations range from20to200μg/mL after incubation for12h, and the effects were significantly higher than that of control group, which was generally better than that of LLF aqueous extract at each corresponding concentration. The promotive effects of Se-LLF alcohol extracts on hybridoma cells proliferation was better than LLF alcohol extracts at the doses of20-400μg/mL24or36h after incubation, which indicates that the aqueous extract and alcohol extract of Se-LLF improve the specific immunity by promoting hybridoma cells proliferation, furthermore,the effects are better than those of LLF.Test5Effect of Se-LLF on antioxidant and immune function of chicken108fourteen days old chickens were randomly divided into three groups, with three repeats of each group,12chickens in each repetition.0.4%Se-LLF aqueous extract was added into drinking water in experimental group, while control group was kept with normal drinking water and fed with the basal diet. At the age of7and21days, the chickens were vaccinated with low virulent live vaccine of Newcastle disease (LaSota) immuning by intranasal and ocular. On day21,28,35and42respectively, Newcastle disease antibody titers and serum interleukin-2level were determinated; at42days of age, GSH-Px, SOD and MDA levels of plasma and tissue and immune organ indexes of chickens were determinated, respectively. The results showed that: Se-LLF can significantly increase the spleen index, the interleukin-2level, the kidney SOD activity, and significantly reduce the chicken plasma and tissue MDA content. The results indicate that addition of0.4%Se-LLF aqueous extract into drinking water can promote chicken antioxidation and non-specific immune function, and Se-LLF has more significant roles compared with the common Ligustrum lucidum.