Reparation And Anti-Cervical Cancer Effect Of Shark Liver Oil

This research paper mainly studied the inhibition effect of shark liver oil on the growth of cervical cancer from 4 aspects: the extraction and evaluation of shark liver oil,the inhibition effect against cervical cancer within shark liver oil, the side effects and the distribution of metabolism in vivo, and the inhibition effect against HeLa cell.Methods: Weak base solution extraction method, Organic solvent extraction method and alkaline protease extraction method were used to extract shark liver oil respectively.Maximize the production rate of shark liver oil by changing the factors such as liquidsolid ratio, dosage of enzyme, acidity, time and temperature of the reaction was studied as well. Squalene, alkoxyglycerols and vitamin A within shark liver oil was characterizated with infrared spectroscopy, gas chromatography and high-performance liquid chromatography(HPLC). SLO was intragastrically administered for 15 days to mice with transplanted cervical carcinoma cells. The mice were sacrificed to assess the tumor inhibition rate, spleen index and thymus index. Mouse serum was collected to test IL-2,IL-4, IFN-γ, superoxide dismutase and malondialdehyde. Tissue homogenates were prepared to measure the antioxidant capacity and oxidative stress levels. Blood, liver,kidney and tumor tissues of the model mice were collected respectively 0.5 h, 1 h, 2 h, 4 h,8 h, 16 h, and 24 h after mice gavaged shark liver oil, the distribution of the shark liver oil within the body of the mice was measureed with the help of HPLC. Normal mice were gavaged with large doses shark liver oil for 30 days, blood samples from the mice’s eyeballs was extracted to mesure liver and kidney contents. The effects of SLO on tumor growth, HeLa cell apoptosis and cell cycle were assessed by MTT method, PI-Hoechst method and flow cytometry, respectively.Results: Among the 4 methods to extract shark liver oil, alkaline protease extraction method was the most effective with 30.1% production rate. Model mice that were treated with excess concentrated shark liver oil result in a rate of inhibition of 54.46%, increased the SOD contents and IL-2/IL-4 ratio of the mice in serum and liver, spleen index was raised as well. Toxicological researches showed that, comparing to normal group, the experimental mice have similar complete blood count results and kidney and liver functions. HPLC test results showed that the t1/2 of squalene and alkoxyglycerols were 2h and 1h respectively. In addition, squalene was majorly distributed in the liver, which suggested that squalene was more attracted to liver; alkoxyglycerol was majorly distributed in the tumor, which suggested that alkoxyglycerol was more attracted to tumor tissues. Shark liver oil(600μg/ml) can inhibit 75% of the HeLa cells growth after 36 h, and it did no particular cytotoxicity to HEK293 T cells in vitro. PI-Hoechst staining results displayed that the number of cells with blue fluorescence was increased by a large amount,while the number of cells with red fluorescence was not increased dramatically. The G1 phase and G2 phase cells were increased from 42.54% and 4.32% to 50.40% and 13.65%respectively, while that of S phase cells decreases from 53.14% to 35.95% which was measured by flow cytometry.Conclusions: The best method to extract shark liver oil was alkaline protease extraction method with a solid-liquid ratio of 2:1, and the dosage of enzyme should be 2%of the mass of the liver. The pH-value, time and temperature were 9, 3.5h, 48 ℃respectively, and maximum production rate was 30.1%. The shark liver oil extracted contains 32.31% squalene, 44.09% alkoxyglycerols and 2.05% vitamin A, contributing78.45% in total. Shark liver oil can effectively suppress the growth of tumors, with a maximum rate of 54.46%. It also improved the immunity and anti-oxidizing abilities. The half-lives of squalene and alkoxyglycerols in shark liver oil were 2h and 1h respectively.Squalene was more attracted to liver, and alkoxyglycerol was more attracted to tumor tissues. Shark liver oil did no obvious harm to the hematopoietic, liver and kidney functions of the mice. Shark liver oil can notably inhibit the growth of HeLa cells with a rate of 76.63%. It induced HeLa cells apoptosis, and the cell cycle was arrested at G1/S and G2/M phase. Therefore, shark liver oil can suppress the growth of cervical cancer.