Entomopathogenic fungus of C. militaris is famous for its medicinal efficacies. It has been reported to have various pharmacological activities such as anti-tumour, insecticidal, antibacterial, immunomodulatory and antioxidant. Many users taking the extracts of C. militaris available on the market a...
Entomopathogenic fungus of C. militaris is famous for its medicinal efficacies. It has been reported to have various pharmacological activities such as anti-tumour, insecticidal, antibacterial, immunomodulatory and antioxidant. Many users taking the extracts of C. militaris available on the market also stress some other positive effects such as in relieving hangovers and in caring the skin, however, few studies have tested these activities of C. militaris. This study is purposing the evaluation of the undisclosed functionalities and the provision of the safety informations of the extracts of C. militaris to make better use of the manufactured functional foods from raw material of C. militaris.
At first, the new strain of C. militaris was searched from various mushrooms growing wild and identified by the analysis of the nucleotide sequences of 5.8S ribosomal RNA. The resulting strain MPUN8501 showed the stable yield of more improved fruit bodies grown upon silkworm pupa. Through the consecutive fractionation and analysis of extract by using of silica column chromatography, thin layer chromatography and high performance liquid chromatography, cordycepin(3`-deoxyadenosine), which is well known effective ingredients of C. militaris, was identified as a major component. Thus it was used as an indicative material for the comparison of various extracting methods. The content of cordycepin was relatively high in hot water and BuOH extractions(CMWE and CME, respectively), which were used for the test of efficacy and toxicity of C. militaris extract.
The effect of CME on the in vitro activities of alcohol dehydrogenase(ADH) and acetaldehyde dehydrogenase(ALDH) were much more than medicine, drink, natural tea etc. CME also promoted the resolution of alcohol and acetaldehyde in alcohol-administered rats, inducing recovery to normal condition rapidly. Furthermore, oral administration of CME effectively protected the carbon tetrachloride-induced acute hepatic injury as revealed by the hematological parameters(levels of sGOT and sGPT) and histological view. CME was ascertained to be safe by regulatory toxicity studies of single dose toxicity and genotoxicity. These results suggest that CME would be useful for the maintenance of hepatic function as a functional health food.
The anti-inflammatory effect of CMWE was performed on LPS-stimulated nitric oxide(NO) production, tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) release in RAW 264.7 cells. The treatment of macrophages with various concentrations of hot CMWE significantly reduced LPS-induced production of NO, TNF-α and IL-6 secretion in a concentration-dependent manner. These results suggest that CMWE have potent inhibitory effects on the production of these inflammatory mediators. In addition, the whitening effect of CMWE was performed with a view of melanogenesis inhibitor. Inhibitory activities of CMWE against tyrosinase, L-DOPA(L-3,4-dihydroxyphenylalanine) oxidation, and melanin biosynthesis in B16 mouse melanoma cells were revealed. The main inhibitory effect of CMWE on melanogenesis was attributed to enhancement of tyrosinase degradation. These results indicated that CMWE may be a potential source of novel whitening agents preventing skin troubles at the same time, which can be applicable for cosmetic or therapeutic uses.
In a separate efficacy and safety studies, water extract of C. militaris grown upon P. dreujtarsis(CMPD) were evaluated. Although P. dreujtarsis has been traditionally used in complementary medicine, its safety was not given definite information. For the efficacy test, SD rats were orally treated with CMPD extract(50, 25 and 12.5 mg/kg body weight) or silymarin(25 mg/kg body weight) before and after administration of CCl4(2 mL/kg body weight, 20% CCl4 in olive oil). Treatment with CMPD extract or silymarin could decrease the GPT(glutamic-pyruvic transaminase) and GOT(glutamic-oxaloacetic transaminase) levels in serum when compared with CCl4-treated group. The results show that CMPD extract can be proposed to protect the liver against CCl4-induced hepatic damage in rats as much as silymarin. The safety tests were designed to evaluate the single dose toxicity and the genetic toxicity - bacterial mutagenicity, chromosome aberration, and micronucleus formation. For single dose toxicity test, the CMPD extract was once administered orally to both sexes of rats at dose of 500, 1000, and 2000 mg/kg body weight which is the recommended maximum limit dose for acute toxicity. Neither significant toxic signs nor death was observed during the observation period. These results indicated that LD50(lethal dose of 50%) of CMPD extract is greater than 2,000 mg/kg body weight in SD rats. For mutagenicity assay, bacterial reversion test with S. typhimurium TA98, TA100, TA1535, TA 1537, and E. coli WP2uvrA were performed, and the extract at the concentrations of 50~5,000 μg/plate did not induce mutagenicity at all. Chromosome aberration test was performed by using Chinese lung(CHL) cells. There was no significant chromosome aberration in CHL cells with S-9 mixture at the concentrations of 312.5~1,250 μg/ml of the extract and without S-9 mixture at the concentrations of 1.2~19.5 μg/ml of the extract. For micronucleus test, ICR mice were treated with the extract at the dose of 0.5, 1, and 2 g/Kg. The frequencies of the micronucleated polychromatic erythrocytes(MNPCE) in bone marrow preparations of the extract-treated group were not increased compared to the untreated control group. Taken together, the results show that CMPD also did not induce any harmful genotoxicity.
Entomopathogenic fungus of C. militaris is famous for its medicinal efficacies. It has been reported to have various pharmacological activities such as anti-tumour, insecticidal, antibacterial, immunomodulatory and antioxidant. Many users taking the extracts of C. militaris available on the market also stress some other positive effects such as in relieving hangovers and in caring the skin, however, few studies have tested these activities of C. militaris. This study is purposing the evaluation of the undisclosed functionalities and the provision of the safety informations of the extracts of C. militaris to make better use of the manufactured functional foods from raw material of C. militaris.
At first, the new strain of C. militaris was searched from various mushrooms growing wild and identified by the analysis of the nucleotide sequences of 5.8S ribosomal RNA. The resulting strain MPUN8501 showed the stable yield of more improved fruit bodies grown upon silkworm pupa. Through the consecutive fractionation and analysis of extract by using of silica column chromatography, thin layer chromatography and high performance liquid chromatography, cordycepin(3`-deoxyadenosine), which is well known effective ingredients of C. militaris, was identified as a major component. Thus it was used as an indicative material for the comparison of various extracting methods. The content of cordycepin was relatively high in hot water and BuOH extractions(CMWE and CME, respectively), which were used for the test of efficacy and toxicity of C. militaris extract.
The effect of CME on the in vitro activities of alcohol dehydrogenase(ADH) and acetaldehyde dehydrogenase(ALDH) were much more than medicine, drink, natural tea etc. CME also promoted the resolution of alcohol and acetaldehyde in alcohol-administered rats, inducing recovery to normal condition rapidly. Furthermore, oral administration of CME effectively protected the carbon tetrachloride-induced acute hepatic injury as revealed by the hematological parameters(levels of sGOT and sGPT) and histological view. CME was ascertained to be safe by regulatory toxicity studies of single dose toxicity and genotoxicity. These results suggest that CME would be useful for the maintenance of hepatic function as a functional health food.
The anti-inflammatory effect of CMWE was performed on LPS-stimulated nitric oxide(NO) production, tumor necrosis factor-α(TNF-α) and interleukin-6(IL-6) release in RAW 264.7 cells. The treatment of macrophages with various concentrations of hot CMWE significantly reduced LPS-induced production of NO, TNF-α and IL-6 secretion in a concentration-dependent manner. These results suggest that CMWE have potent inhibitory effects on the production of these inflammatory mediators. In addition, the whitening effect of CMWE was performed with a view of melanogenesis inhibitor. Inhibitory activities of CMWE against tyrosinase, L-DOPA(L-3,4-dihydroxyphenylalanine) oxidation, and melanin biosynthesis in B16 mouse melanoma cells were revealed. The main inhibitory effect of CMWE on melanogenesis was attributed to enhancement of tyrosinase degradation. These results indicated that CMWE may be a potential source of novel whitening agents preventing skin troubles at the same time, which can be applicable for cosmetic or therapeutic uses.
In a separate efficacy and safety studies, water extract of C. militaris grown upon P. dreujtarsis(CMPD) were evaluated. Although P. dreujtarsis has been traditionally used in complementary medicine, its safety was not given definite information. For the efficacy test, SD rats were orally treated with CMPD extract(50, 25 and 12.5 mg/kg body weight) or silymarin(25 mg/kg body weight) before and after administration of CCl4(2 mL/kg body weight, 20% CCl4 in olive oil). Treatment with CMPD extract or silymarin could decrease the GPT(glutamic-pyruvic transaminase) and GOT(glutamic-oxaloacetic transaminase) levels in serum when compared with CCl4-treated group. The results show that CMPD extract can be proposed to protect the liver against CCl4-induced hepatic damage in rats as much as silymarin. The safety tests were designed to evaluate the single dose toxicity and the genetic toxicity - bacterial mutagenicity, chromosome aberration, and micronucleus formation. For single dose toxicity test, the CMPD extract was once administered orally to both sexes of rats at dose of 500, 1000, and 2000 mg/kg body weight which is the recommended maximum limit dose for acute toxicity. Neither significant toxic signs nor death was observed during the observation period. These results indicated that LD50(lethal dose of 50%) of CMPD extract is greater than 2,000 mg/kg body weight in SD rats. For mutagenicity assay, bacterial reversion test with S. typhimurium TA98, TA100, TA1535, TA 1537, and E. coli WP2uvrA were performed, and the extract at the concentrations of 50~5,000 μg/plate did not induce mutagenicity at all. Chromosome aberration test was performed by using Chinese lung(CHL) cells. There was no significant chromosome aberration in CHL cells with S-9 mixture at the concentrations of 312.5~1,250 μg/ml of the extract and without S-9 mixture at the concentrations of 1.2~19.5 μg/ml of the extract. For micronucleus test, ICR mice were treated with the extract at the dose of 0.5, 1, and 2 g/Kg. The frequencies of the micronucleated polychromatic erythrocytes(MNPCE) in bone marrow preparations of the extract-treated group were not increased compared to the untreated control group. Taken together, the results show that CMPD also did not induce any harmful genotoxicity.
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