Isolation Of Dairy Goat Placental Peptide And Effect On Immune Function Of Mice

In1985, Liu Yuexin reported that the placental peptide was extracted by"homogenized - dialysis"for the first time . physics and chemistry analysis and the biological activity on placental peptide were conducted, discovering that it used in clinical treatment of the virus Hepatitis, leukopenia, myasthenia gravis, and other autoimmune diseases ,cancer and so on,. The structure and the function of dairy goat placental has many similarities with humen's placenta,In the last few years ,some reports it was possible to extract the goat placenta peptide from goat's placenta, and has similar biology activeness with the human placenta peptide.There is not report on the physics and chemistry analysis and the biological activity of dairy goat placental peptide , looking at the human placenta and whether they are consistent with the activity of peptide..it maybe provide certain basis for the dairy goat placental peptide research and it uses in treating the animal or the person diseaseMore than the previous report focused on the system the mix small peptide active research. by the dialysis or ultra filters.This experiment will pass through ultra filters to mixes the small peptide with the superdex75 chromatographic analysis to carry on the separation, through examines each component biology activeness to screen the activated component, provides certain basis for the next step biosynthesis or the chemical synthesis. The study consists of two parts:1. Isolution and content determination of dairy goat placental peptideThe freezing dairy goat placenta normal temperature defrosts, after cleaning the blood, with the physiological saline flushing placenta, takes placenta's seed leaf, the seed leaf cuttings, passes through repeatedly the freezing and thawing, after the high speed organization stamp crushing refiner refining, freezes the centrifuge offcenter to precipitate high speed, is accepted after passing an examination the serum uses separately 0.45μm, 0.22μm the filter to carry on micro-filters, by except insoluble substance The micro filtrate surpasses the filter filtration after Millopor YM-10 to obtain the thick sample, the thick sample is the colorless transparent liquid, has may the dialysis, but ultra filters the nature, the non-pyrogen's small molecular multi-peptides, the pH is6.6~6.9.Crude sample obtains four component peptides class after the superdex75 chromatographic analysis, respectively isⅠ,Ⅱ,Ⅲ,Ⅳ, after freeze-drying, is the white powder.The four components after add the urea and the glycerine Tricine-SDS-PAGE electrophoresis electrophoresis. Result demonstration componentⅠ,Ⅱ,Ⅲ,ⅣThe molecular weight respectively is 10.139KDa, 8.166KDa, 7.447KDa and 6.761KD.By Coomassie brilliant blueblue G-250 determination componentⅠ,Ⅱ,Ⅲ,ⅣThe content is: 0.96mg/g, 1.2mg/g, 1.32mg/g, 1.8mg/g.2. Dairy goat placental peptide on immune function in miceThis experiment divides into stochastically the mice 6 big groups, 5 groups for experimental group (distinction abdominal cavity injection component:Ⅰ,Ⅱ,Ⅲ,ⅣWith raises the crude sample ); Each experimental group also divides into the high,school,low three dosage group (respectively is 0.1mL, 0.2mL, 0.4mL), each group 5; Control group 5, only inject the physiological saline。Every day injects one time, injects continuously for one week. Through to the mice circumference blood serum Bai IL-2 content, the spleen T lymphopoiesis response, spleen NK cell activeness and the abdominal cavity macrophage swallow function's influence inspects the dairy goat placenta peptide the immunity activeness function.1.determines in the mice circumference blood through the radioactive immunization the IL-2 content, the results show that componentⅢ,Ⅳand crude sample can be significantly increased in the peripheral blood of mice IL-2 level, than the control group increased 15.3 % ~ 30.1% (p <0.05). 2.Determination by MTT spleen T-lymphocyte proliferation, the results show that componentⅢ,Ⅳand crude sample in the body can be very significant synergies ConA stimulate the spleen lymphocyte proliferation (p <0.01) .3.Determination by MTT spleen of anti-NK cell activity, showed that componentsⅢ,Ⅳand crude sample can significantly improve the mouse spleen NK cell killing activity (p <0.01).4.Peritoneal macrophages through phagocytosis neutral red experiment the results show that componentsⅢ,Ⅳand crude sample can significantly improve mouse peritoneal macrophage phagocytosis (p <0.01).By the above dairy goat placental peptide to the mice immunologic function influence's 4 experimental result, discovered that the existence dosage relations, can add along with the density strengthen, but the difference is not remarkable; Has the immunity actively what in the dairy goat placenta peptide is the componentⅢandⅣ.