Study of the Extraction Process and In Vivo Inhibitory Effect of Ganoderma Triterpenes in Oral Mucosa Cancer

Abstract

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.

Extraction of triterpenoids and phenolic compounds from Ganoderma lucidum: optimization study using the response surface methodology

“…UA per g dw was obtained. These claims have been supported by other authors 36 who reported that the extraction of Tr from G. lucidum is largely influenced in the T range between 80 and 100°C and if prolonged time-cycles of 2 h (∼4 cycles) are used.…”

Applied modern biotechnology for cultivation of Ganoderma and development of their products

“…The different extraction methods of Lingzhi triterpenes mainly include (1) extraction with methanol or ethanol as the solvent, followed by direct separation of the extracts; (2) extraction by methanol or ethanol, followed by separation of the total acid portion by alkali treatment and separation of Lingzhi triterpene; and (3) extraction of the total acid portion by ether, followed by diazomethane methylation and then separation. Extraction of Lingzhi triterpene using ethanol is the easiest approach to maintain the activity of the extracts and to scale up its production (Gao et al 2011). In addition, in order to enhance the rate of extraction of triterpenes and shorten the extraction time, the ultrasonic technique was often used for treatment of raw materials (fruit body or spore) in the production, which would destroy the dense structure of Lingzhi cells.…”

Hepatoprotective effect of ganoderma triterpenoids against oxidative damage induced bytert-butyl hydroperoxide in human hepatic HepG2 cells

“…The total triterpenoid content was determined according to the report previously with some modification (Gao et al 2011), by using ultraviolet-visible spectrophotometry (UV-1800 UV/VIS spectrometer, Shimadzu, Tokyo, Japan) with ganoderic acid A as the control and 5% glacial acetic acid-vanillin solution and perchloric acid as the developer. The total triterpenoid content in GTs was calculated as milligrams of ganoderic acid A equivalents (GAAEs) per gram of GTs.…”