The aim of the reported study was to optimize the extraction process for ganoderma
triterpenes and to investigate the in vivo inhibitory effect of ganoderma
triterpenes on the genesis and progression of
oral cancer. Single-factor and orthogonal methods were used to investigate the effects of extraction
solvent,
solvent amount, extraction time, extraction temperature, and number of extractions, on the extraction rate for ganoderma
triterpenes. A golden hamster model with cheek pouch dynamic canceration was established to receive oral treatment of ganoderma
triterpenes water
solution. Animals were continuously monitored, oral tissue samples were collected for histopathologic examination, and changes in the expression of
VEGF (
vascular endothelial growth factor) and
Caspase-3 were detected by immunohistochemical methods. Optimization of the experimental conditions allowed the identification of the optimal extraction conditions: 90%
ethanol as the extraction
solvent, a
solvent amount by the liquid-material ratio of 35 mL/g, extraction time of 2 h and extraction temperature of 80 °C. Under these conditions, the average extraction rate of ganoderma
triterpenes was 1.09%. Tests in golden hamsters showed that compared with the model group during the same period, animals in the treatment group had better conditions, constantly larger number of normal cases shown by histopathologic results (P < 0.01), and consistently smaller numbers of cases with paraplasm (P < 0.05). Immunohistochemical results showed that compared with the model group, the treatment group had significantly lower (P < 0.05) rates of positive
VEGF expression in the normal state, simple epithelial
hyperplasia, epithelial dysplasia or
squamous cell carcinoma disease stages.
Caspase-3 expression showed a tendency toward a gradual increase with the worsening of disease severity in each group. Compared with the model group, the treatment group had significantly lower (P < 0.05) rates of positive
Caspase-3 in the normal state, simple epithelial
hyperplasia, epithelial dysplasia or
squamous cell carcinoma disease grades. Using the optimized extraction process, ganoderma
triterpenes could be extracted with high efficiency, and the results of animal tests showed inhibitory effects of ganoderma
triterpenes on oral mucosa
cancer.