| With the purpose to develop the soft-shelled turtle health care products, separated and purificated vitamin B-17 from soft-shelled turtle in the experiment. The studies were used vitro and vivo tests to discuss the effect of vitamin B-17 to HepG2 cells growth, and analysed biochemical, immunological and pathological factors, to confirm the effcts of vitamin B-17 to the growth, immunization and pathology of the tumor-bearing mice, explored the anti-tumor mechanism of vitamin B-17.1. Using HPLC to separate and purificate vitamin B-17 from soft-shelled turtle powder, got the pure of vitamin B-17.The results show that it can separable purifie 10.39mg vitamin B-17 per 1kg soft-shelled turtle protein powder, detected by HPLC, the extraction purity reaches 90.40%.2. Using MTT to examin the effects of differdnt concentrations of vitamin B-17 to the growth of HepG2 cells in vitro tests. The results show that in a certain concentration range, addition of vitamin B-17 and HepG2 cell growth inhibition is positively correlated. In vitro tests, when the dosage of vitamin B-17 is 1.3mg/mL, HepG2 cells growth inhibition rate is 5.62%; when the dosage was 10.5mg/mL, the cells inhibition rate has increased to 30.99%; and when adding dosage is 42mg / mL, the inhibition rate has reached 56.30%.3. HepG2 cells inculated tumor-bearing mice were fed with defferent concentrations of vitamin B-17, to examined the effects of vitamin B-17 to the growth of HepG2 cells in vivo tests. The results show that in vivo tests, adding 15mg/mL and 30mg/mL vitamin B-17 has no distinct effects to tumors'growth inhibition, the growth inhibition rates are only 14.73% and 31.95%; 60mg/mL vitamin B-17 group can restrain tumor growth obviously, inhibition rate is 52.49%. As growth inhibition, 60mg/mL vitamin B-17 group has restrained the growth of tumor-bearing mice too. Compared with the control group, ADG has reduced 1.07 times(P<0.01).4. Using HE staining and pathological staining, studied the effects of different concentrations of vitamin B-17 to the pathology of HepG2 cells inculated tumor-beating mice. The results show thar The addition of vitamin B-17 had a certain influence to the tumors. As the concentrations of vitamin B-17 increased, the tumor positive expression rate of Survivin and VEGF decreased. When vitamin B-17 concentration is 15mg/mL, the tumor positive expression rates of Survivin and VEGF have reduced 21.8%(P>0.05) and 13.64%(P>0.05); when adding concentration is 30mg/mL, Survivin and VEGF positive expression rates have reduced 30.4%(P<0.05) and 28.31%(P>0.05); but when the adding concentration have increased to 60mg/mL, Survivin and VEGF expression rates have reduced 35.60% (P<0.05) and 50.83%(P<0.01).5. By analyzed the serum and liver-related physiological changes, to stueied different concentrations of vitamin B-17 to the physiological metabilisn of tumor-bearing mice.The results show that the addition of vitamin B-17 had a certain influence to pathology of tumor-bearing mice. Compared with the control group, serum protein contents in 60mg/mL vitamin B-17 adding group has decreased by 17.77% (P>0.05), LPL has decreased by 65.08% (P<0.01), NEFA has decreased by 15.80% (P>0.05), serum ALB has increased by 16.19% (P>0.05). And TC,HL and MDH in 60mg/mL vitamin B-17 adding group have increased by 85.56% (P<0.01),58.60%(P<0.01) and 22.99%(P<0.05).6. By analyzed the serum immune parameters, to studied defferent concentrations of vitamin B-17 to the immune of the tumor-bearing mice.The results show that the addition of vitamin B-17 can invigorate immunity effectively of tumor-bearing mice. The serum IgA in 15mg/mL and 30mg/mL group have increased by 68.81%(P<0.05) and 1.09 times(P<0.05); Compered with control group, the difference of serum IgG in 15mg/mL group is not obvious(P>0.05), 30mg/mL group have increased by 23.88% (P<0.05), The serum IgA, IgG in 60mg/mL vitamin B-17 adding group has increased by 225.25% (P<0.01) and 38.47% (P<0.05) to control group. |
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