Detection Of Vitamin B₁₇ From Turtle And The Research Of Anti-Hepatic Carcinoma Activiy Of Vitamin B₁₇ Following Specific Activation Byβ-glucosidase

The ultrasonic extraction technology was used in the study for extracting vitamin B17from turtle, and then qualitative and quantitative analysis by HPLC. Research of anti-hepatic carcinoma effect of vitamin B17following specific activation by β-glucosidase on human hepatic cancer HepG2cells in vitro and in vivo, and to analyze its possible mechanism, offering experimental data for exploring the turtle health care products.1.Using HPLC to analysis the extracting solution from turtle. The results showed that:under the conditions of ethanol concentration at80%, solid-liquid ratio1:15, the dextration time30min under20℃, it contained10.46mg vitamin B17per kilogram turtle.2. Culture human hepatic cancer HepG2cells in vitro to study the cytotoxicity of vitamin B17fowllowing specific activity by (3-glucosidase. MTT assay results:the inhibition rate was observed when separately administered vitamin B17or continuously with β-glucosidase, but the inhibition rate of koinonia was higher than the separately administered, with the increase of vitamin B17concentration and the incubation time, the inhibition was reinforced. Observeing under the microscope:With the increase of vitamin B17concentration and the incubation time, cells transformed to round, crimple, the numbers of adherent cell were fewer.14mg/mL group already presented the form of necrosis. AO/EB fluorescent staining method detected:with the increase of vitamin B17concentration and the incubation time, apotosis number was increased. Compared with the control group, there was significant difference (P<0.01). Annexin V-FITC/PI staining FCM assay: with the increase of vitamin B17concentration and the incubation time, apotosis rate was heightened. Compared with the control group, there was significant difference(P<0.01).3、Setting up the subcutaneously transplantable tumor model of human hepatic cancer HepG2cells to investigate the effects of vitamin B17on tumor growth in vivo. The results showed that:the relative tumor volume and tumor weight of the high(60mg/mL), medium(30mg/mL), low-dose(15mg/mL) intragastric administration group and intraperitoneal injection(60mg/mL) group, all smaller than the control group, the inhibition rates of the four groups were58.30%,45.64%,5.81%and14.73%, respectively, medium and high-dose groups compared with control group were statistically significan(P<0.01), However, could be seen from the inhibitory effects of high-dose intragastric administration group better than the same dose intraperitoneal injection obviously (P<0.01).4、Observing the behavior and daily weight gain of nude mouse, to analyze the toxic and side-effect that caused by intragastric administration and injection vitamin B17. The results showed that:Low-dose intragastric administration group and intraperitoneal injection group compared with the control group did not show significant difference (P>0.05). The daily weight gain of high-dose intragastric administration group decreased94.09%(P<0.01) compared with the control group, thus compared with the intraperitoneal injection group, it decreased93.66%(P<0.01). Medium intragastric administration group compared with the control group、intraperitoneal injection group and low-dose group, the daily weight gain decreased15.69%,20.15%and23.21%(P<0.05), respectively.