| Objective:In recent years, liver cancer prevention has aroused more and more attention with the increasing rate of the liver cancer. Codonopsis lanceolata (CL)is a kind of medical herbal root(now used as a kind of food), which composed of many kinds of nutrient substances.Reported in the literature, CL have a preventive role in alcoholic liver injury, antioxidant and anti-mutation, anti-tumor effect, but fermented codonopsis lanceolata (FCL) have no inhibition of liver cancer yet.This experiment tries to strain screening to ferment from stinky of CL, best condition optimize, studies effective parts of inhibit liver cancer and provide academic foundations for chemoprevention of live cancer preparation.Method:(1)The oleanolic acid, total saponin and total flavonoid in the CL fermented with yeasts, acid bacterium, EM bacterium were determined by colorimetric method and ultraviolet spectrophotometry method to compare with unfermented codonopsis lanceolata. (2)Transplanted H22 tumor-bearing mice were used as a model for observing the anti-tumor effects of extracts from fermented and unfermented CL: The mice were divided into three groups and given each type of extract, with receiving a high dose group(220 mg/kg bw), medium dose (110 mg/kg bw) and low dose group(55mg/kg bw), model group was given distilled water (10mL/kg), cyclophosphamide group (intraperitoneal injection of cyclophosphamide 25 mg/kg bw), and on alternate days, spleens and thymuses were taken and weighted after 10 days. (3)Screening for antiviral anti-liver cancer from FCL:FCL of petroleum ether, ethylacetate, n-butanol portions to antihepatocellular-carcinoma was screened in codonopis lanceodata. Inhibits the proliferation of hepatocarcinoma cell H22 was observated for these portion. (4)The application of orthogonal design method to optimizing yeasts dose, fermented times, glucose dose, fermented temperature. (5)The inhibitory effect of fermented and unfermented CL n-butanol part on tumor growth inhibition and apoptosis of hepatoma cells:A study on comparison of MTT for fermented and unfermented CL n-butanol part was observed by the models of H22 and hepatoma carcinoma HepG2 in vivor.Result: (1) After CL was fermentated for yeast, the content of oleanolic acid and tatal saponin is highest; FCL oleanolic acid and tatal saponin are higher than unfermented CL (P<0.05). The content of total saponins is unfermented CL 2.1 times. (2) FCL middle and high dose groups for the weight of tumor is markedly lower than the model group (P<0.01), the middle dose group inhibition rate by more than 50%. The tumors weight in unfermented CL low dose group and middle dose group is obviously lower than model group. The inhibition rate of middle dose group more than 30 percent. But three doses groups were not more than 50% inhibiting rates. Fermented and unfermented CL groups spleen index,thymus index was higher than the cyclophosphamide groups, compared with model group. There was no significant difference. (3) Proliferative inhibition of hepatocellular carcinoma cells by FCL petroleum ether (500μg/mL), ethylacetate (500μg/mL,250μg/mL), n-butanol (500μg/mL,250μg/mL) portions is evident higher than solvent control group (P<0.05 or P<0.01). Among them, n-butanol (500μg/mL,250μg/mL) portion has the highest inhibiting rate (P<0.01). (4) The optimum fermentation conditions were 1%yeast, 5%dosage of glucose,24h fermentation time was strongest inhibited the rate (P<0.01). (5) FCL 400μg/mL and 600μg/mL groups for inhibits the proliferation of hepatocarcinoma cell H22 was higher than unfermented CL in vitro (P<0.01). FCL 400μg/mL for inhibits the proliferation of hepatocellular carcinoma cell HepG2 was higher than unfermented CL in vitro (P<0.01).Conclusion:1. To promote total saponin the best advantages was fermentation by yeast of CL.2. Antihepatocellular-carcinoma for fermentation of CL was better than unfermented.3. The n-butanol portion has the strongest effect of anti-hepatocellular-carcinoma.4. Best conditions for fermented of CL:0.5%-5.0%yeast,1%～8%glucose, 1～10days fermented time. |
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