| Fig(Ficus caraca L.),also known as "Tian Shengzi",is a plant of the Ficus genus in Moraceae family.The fig leaves are rich in a variety of biologically active ingredients such as flavonoids,volatile oils,phenolic compounds,polysaccharides and alkaloids.As the most important active ingredient from the fig leaves,polysaccharide has potential biological activities such as anti-oxidation,anti-virus and anti-tumor.However,there are few studies on fig leaves polysaccharides.Fig leaves were used as raw material to purify polysaccharide in this study.Furthermore,the anti-tumor activity of fig leaf polysaccharides(FCPS)in vitro and in vivo was studied.The main results are as follows:(1)By optimizing the response surface method of the extraction process parameters of FCPS,such as liquid-solid ratio,microwave power and extraction time,the optimal ultrasonic-microwave assisted extraction(UMAE)process parameters were obtained by software model fitting:the ratio of material to liquid was 31 mL/g,extraction time for 15 min,the microwave power is 540 W.Experimental results showed that under the optimal extraction parameters,the optimal yield of FCPS were 4.52±0.12%.In addition,the UMAE was compared with traditional hot water extraction,ultrasonic extraction and microwave extraction.The results showed that the UMAE can obtain the highest yield of FCPS in the shortest time.Compared with the traditional hot water extraction method,the extraction time of UMAE is decreased by 83.3%,and the extraction yield of FCPS was increased by 46.9%.The UMAE has the advantages of short extraction time and high yield for extracting of FCPS.(2)It was determined that the protein in the FCPS was removed by the Sevag method.Adsorption experiments were carried out on the extracts of FCPS with six kinds of macroporous resins AB-8,D101,HPD-826,HPD-400,HPD-100 and NKA-9.It was proved that D-101 resin has strong adsorption capacity for impurities other than polysaccharide.The purpose of removing pigments and small molecular impurities was achieved.The FCPS separation conditions using cellulose column chromatography(DEAE-32)were determined.The yield of FCPS was 1.52%and its purity was 91.45%.(3)The human lung cancer cell A549,human cervical cancer cell Hela and human hepatoma cell HepG2 were used as test cells,and the cytotoxicity of FCPS was explored by MTT assay.The results showed that FCPS had the weak cytotoxicity to the three tested cells.Comparatively,FCPS had the highest inhibition yield on human cervical cancer Hela,followed by human hepatoma cell HepG2,but human lung cancer cell A549 was not sensitive to FCPS.The IC50 of FCPS inhibiting the growth of human cervical cancer cells and human hepatoma cells HepG2 were 5.82 mg/mL and 6.65 mg/mL,respectively.The inhibitory effect of FCPS on S180 tumor-bearing mice was manifested in the changes of tumor weight and body weight in mice,and its inhibitory effect was concentration-dependent.When the FCPS administered dose were 400 mg/kg,the tumor inhibition effect is the largest,and the tumor inhibition rate is 51.82%.The effect of FCPS on the relative coefficient of thymus and spleen organs in mice,the thymus and spleen coefficients of the three polysaccharide-administered groups increased,showing the repair of thymus and spleen damage to the tumor,and there was no significant difference between the dose groups.The effect of FCPS on lymphocyte proliferation in mice and the levels of TNF-? and IL-1? in serum of mice showed that high dose of FCPS can significantly enhance the proliferation of lymphocytes,and FCPS in both medium and high dose groups can significant increased TNF-a and IL-1? levels in mice serum.In summary,FCPS inhibits tumor cells primarily by increasing mouse immunity rather than by its cytotoxicity. |
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