Research On The Inductive Methods And Preparative Conditions Of Rabbit Liver Zinc Metallothionein (MT)

Objects: In order to improve the MT produce,research the inductive method of synthesizing MT in rabbit liver and the best technique extracting MT from it and method of separating and 'purifying MT.Methods: 1.Induce MT by subcutaneous injection of zinc sulphate and dexamethasone sodium phosphate in rabbits and detect the contents of zinc in the rabbits liver by atomic absorption spectrophotometry(AAS).2.Use ultraviolet spectrophotometry(UV) and AAS to mensurate sulfhydryls' activity and Zinc content and use them as markers to dectect MT of extracted liquid; Adopt an orthogonal design of L9(34) to research the best extracting technique.Use gelatum chromatography and anion exchange chromatography to separate and purify MT.3. Research the purity and the molecular weight of MT with liquid chromatography-mass spectrophotom-etry;Measure the content of MT with UV and AAS.Results: 1. The inductive method can inprove the MT of the rabbit liver markedly and Znic sulphate and dexamethasone have significant synergistic action(p < 0.01).2.Obtaining a suitable technique to extractMT.3. Obtain two type of MT(MT-I and MT-II) with chromatography and get it molecular weight is 6500D. MT-I and MT-II not be separated by RHPLC with gradient elution.4.When separating MT with gelatum cbromatography,with NH4HCO3 replacing Tris,the produce can be lyophilized but don't need to desalt. It simplifies the procedur.5. Measure the produce rate of MT of rabbit liver which is be induced by s.c. five times Znic sulphate is 2.9mgMT/g wit liver,and the purity of MT is about 90%.Conclusions: 1. Znic sulphate and cortex hormone induce together can increase the synthesis rate of MT.2. The concentration , temperature dosage and proportion of ethanol and water of extract liquid all influence the extract rate of MT.3. When separating MT with gelatum chromatography. NH4HCO3 can replace Tris.4. The molecular weight of MT of rabbit liver in this experiment coincide with literature.5.This method of preparing MT fit laboratory preparation.