| Lead poison, which can influence the function and life of erythrocyte, lead to the derangement of begetative nerve and hurt new protein composition and cranial nerve of key function for forming long-lasting memory, damage the function of renal tubule, and so on. Nowadays, there are two major clinical and health care remedial ways for curing lead poisoning. The experiment will develop and use homologous material of natural medicine and food to study functional compound recipe liquid to eliminating lead, which is of the more important preventing significance for people who have a certain lead accumulation in body, while not reaching poisoning content. Basing on studying preparation and purification of bean antioxidant peptide, the study is mainly aiming at the efficacy factor of compound functional oral liquid to establish mathematical model, evaluate economic character and fuzzily evaluate its efficacy, which provides the more scientific theoretical support in nutrition and healthcare. Researching content is as followed:1. Clone and expression of antioxidant peptide pET-22b(+)-apr recombinant strain of using efficacy enzymolysis soybean protein. Bacillus subtilis protease apr was successfully cloned by PCR method and recombinant expression plasmid pET-22b (+)-apr was constructed and expressed in E.coli BL21(DE3), which full-length of gene sequence was 1509bp, 503 amino acid being coded, protein molecular weight being 55 ku, which the consistency of sequencing results was 100% with implementation sequence. Expression protein existed in inclusion body at 37℃.2. The research of separating soybean antioxidant peptide with gel filtration chromatography. Using sephadex G-25 gel filtration chromatography to separate soybean hydrolysates, Four kinds of peptide components were obtained: SP1 in 78266Da above, SP2 of the range of 13240 ~ 78266Da, SP3 of the range of 1439 ~ 1067Da, SP4 of 156 ~ 1439Da. The anti-oxidation activity of these four kinds of peptide components was measured. The results showed that the antioxidant activity of SP4 was strongest. Through the amino acid composition analysis, peptide SP4 was composed of 16 kinds of amino acids. SP4 contained three peptide peaks with reversed-phase HPLC analysis.3. The research of functional compound recipe liquid to eliminating lead design experiment and exterior-body functional appraisal. Through simulating stomach condition in exterior-body, the better ratio range and different dose chelated effect of every efficient elements were determined previously and every efficient elements'chelated effect in various doses were compared, then five factors A,B,C,D,E being obvious effect were studied. With penta-factorial quadratic orthogonal experimental design, the research combined the economic character of compound eliminating lead oral liquid and fuzzily appraised five factors of the oral liquid edible performance to determine the best combination: Gly being 1.46g, mixed extracted solution of Chinese medicine being 1.09mL, soybean antioxidant peptide being 0.94g, Zn being 0.33g and Ca being 0.17g.4. Study on the eliminative functional compound recipe liquid to eliminating lead used to eliminate lead. 120"Kunming"rats being 18～20g were divided into 6 groups according to weight: blank group(BD), negative group(ND), positive group(PD) and high group (HD,6.67mg/kg.bw.d ),middle group (MD,3.33mg/kg.bw.d and low group(LD,1.67mg/kg.bw.d three dose groups). Except the BD, others groups drunk 0.24% Pb(AC)2, filled stomach with experimental liquid and DSMA. To 20d, blood were collected from tail once; to 40d, limosis 24h, liver, thighbone and blood were collected to determine the content of microelement: Pb2+, Ca2+, Zn2+. The results were as followed: lead content difference in plasma was extremely significant comparing the three dose containing eliminative lead compound oral liquid with BD, ND and PD(P<0.01); the diversity of lead content in liver and bone was not significant comparing MD with BD; The difference of HD bone lead content was not significant with PD. Eliminating lead compound oral liquid can supplement the good microelement Ca2+ and Zn2+ in body. The results proved that eliminating lead compound oral liquid had extremely difference with BD, ND and PD(P<0.01,P<0.05).5. Study on the antioxidative function of functional compound recipe liquid to eliminating lead. 75"Kunming"rats being 18～20g are divided into 5 groups according to weight: BD,MoD,HD ( 6.67mg/kg.bw.d ),MD ( 3.33mg/kg.bw.d ) and LD (1.67mg/kg.bw.d). Freely drunk, filled stomach with experimental liquid, to 4 weeks end and limosis 10h, then the SOD and GSH-PX activity in plasma and MD content in plasma and liver were determined. The result were as followed: three dose groups of eliminating lead drink took on strong antioxidative function, MDA content in plasma and liver being significantly decreased(P<0.01), SOD and GSH-PX activity being significantly increased (P<0.01).
|
PDF Full Text Request