Flavone Extraction From Tibetan Saxifraga Tangutica And Studies Of Its Antitumor Effects

Saxifraga tangutica Engl.,a Tibetan medicine,was grew in coniferous forests shrubs at altitude of 2900-4900 m in Qinghai,Gansu,Tibet and Sichuan of china.It tastes bitter and cool-natured,purging liver and gallbladde,healing wounds,treating acute otitis media,wind-heat cough.Pharmacological studies have shown that Saxifraga tangutica Engl.has antibacterial,antiviral,anti-inflammatory and anti-tumor effects.Saxifraga tangutica Engl.is rich in bioactive substances such as flavonoids and polysaccharides,and has great value in development and application.We conducted the study as follows:1.Extraction and purification of flavonoids from Saxifraga oleracea.Firstly,the crude flavonoids of Saxifraga tangutica by ultrasonic-assisted ethanol extraction,which followed by extraction with ethyl acetate,the purity of flavonoids in the extract is 48.5%.Finally,The crude flavonoids of the extract was further purified by NKA-II macroporous adsorption resin,and the purity of flavonoid obtained was up to 82.07%.Based on the single factor test,the orthogonal test was designed to optimize the extraction process:ethanol 80%,material-to-liquid ratio 1:25 g/mL,extraction temperature 50 ^oC,extraction time 35 min,and extraction number 4 times.The extraction volume could reach 81.73 mg/g.2.Studies on the anti-tumor effect of saxifraga flavonoids extracted by ethyl acetate.Co-incubation of saxifraga flavonoids with mouse H22 hepatocarcinoma cells can induce apoptosis.Nucleic acid electrophoresis showed a characteristic ladder-shaped band of apoptosis.Analysis of flow cytometry showed that the apoptosis rate reached 31%.MTT assay was used to determine the inhibition rates,It was positively correlated with the action time and concentration?P<0.05?.H22 was injected into the liver of mice to establish a model of orthotopic transplantation tumor.A significant inhibitory effect of Saxifrage flavonoid on orthotopic transplantation of mice was oral administration.The tumor inhibition rates of the low,medium and high dose groups were 33.0%,46.7%and 64.3%respectively.The survival time was extended by 13.11%,33.01%and 46.12%respectively.The levels of tumor marker enzymes?AFU,ALP and GGT?,the liver function indexes?ALT and AST?,and the liver MDA content decreased significantly?P<0.05?.However,ALB,A/G ratio,the activity of GSH-Px and the value-added ability of the T lymphocytes increased dramatically?P<0.05?.The maximum of the inactivation of the disinfectant on the acute toxicity test was LD50>10000mg·kg^-1.The results showed that saxifraga flavonoids extracted by ethyl acetate was practically non-toxic.Conclusion:saxifraga flavonoids extracted by ethyl acetate has anti-hepatocellular carcinoma effect.