Box-Behnken design criterion was applied to identify the significant effects of various extraction parameters such as temperature, time, and solvent-solid ratio on extraction of tea carbohydrate. Among the three variables tested extraction temperature, and solvent-solid ratio were found to have significant effect on tea carbohydrate extraction. The most suitable condition for extraction of tea carbohydrate was found to be a single step extraction at extraction temperature 90°C, extraction time 30 min, and solvent-solid ratio 5:1. At these optimum extraction parameters, the maximum yield of tea carbohydrate obtained experimentally was found to be very close to its predicted value of 3.47% dry weight of root. Then, we have studied the influence of tea carbohydrate on biochemical parameters in hepatocellular carcinoma (HCC) animals. Hepatocellular carcinoma was induced by the injection of 1×10(5) H22 hepatocarcinoma cells into right hind thigh muscle in experimental animals. Tea carbohydrate could inhibit tumour growth and decrease microvessel density in tumour tissue. The altered amount of serum white blood cells (WBC), Interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) in HCC animals were dose-dependently increased, whereas activities of serum alanine transaminase (ALT), aspartate transaminase (AST) and alkaline phosphatase (ALP) were dose-dependently decreased in the drug treated animals. In addition, tea carbohydrate administration could decrease expression of vascular endothelial growth factor (VEGF) and proliferating cell nuclear antigen (PCNA) in H22 tumor tissue. It can be concluded that tea carbohydrate displayed strong antitumour activity in animals.
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