Artificial Culture Of Cordyceps Sinensis, Component Analysis And Its Hypoglycemic Effect Studies

Cordyceps sinensis is one of the hotspots of modern medicine. The cultivation , component analysis and pharmacological studies of Cordyceps were investigated in this paper.The seed production techniques of C. sinensis were explained by spore separation and tissue isolation methods. In both methods, the steps were operated as following : the expansion of Mother Culture, Preparation of suspension, liquid bacteria, technical of cultured mycelium and breeding of bacteria and preservation. The result showed that parent species of C. sinensis could be isolated by spore separation and tissue isolation methods. The tissue isolation was more simple and had higher success rate. At last the artificial C. sinensis was cultured with rice as its medium.To determine the medicinal ingredients in Cordyceps, the funguses polysaccharide, mannitol, and adenosine in C. sinensis were investigated. Anthrone sulfuric acid method and phenol sulfuric acid method can be used to determined C. sinensis'polysaccharide. The results showed that with anthrone sulfuric acid method, the content of C.sinensis'polysaccharide of wild C. sinensis , artificial Cordyceps and wild Cordyceps mannitol were 5.44%, 5.77%, 20.55%, respectively; while with the phenol sulfuric acid method, the content were 5.46%﹑5.65%, 16.86%, respectively.The content of mannitol in Cordyceps was determined by Sodium thiosulfate titration method. The result showed that C. sinensis and C. militaris had a higher content of mannitol: 8.93 mg/g and 8.08 mg/g, respectively; but the dried artificial C. mycelia's mannitol was only half of them, which was 4.02 mg/g.Thin Layer Chromatography Analysis was used to study Cordyceps'Adenosine. The results showed that Rf values of C. sinensis, C. sinensis and C. mycelia were 0.70, 0.70, 0.75, respectively; and because the Rf of adenosine standard was 0.71, then a conclusion could be made that all these three glycosides contained glands. The preliminary results show that the artificial culture of Cordyceps has the potential to be the drug source.To lay the foundation for DNA fingerprinting technology to identify the authenticity of Cordyceps, CTAB, SDS and Urea methods were used to extract C. sinensis'total DNA. The results showed that dried C. sinensis contained more phenol and less sugar,therefore CTAB extraction method was more suitable, while fresh C. sinensis contained less phenol and more sugar, so Urea method was more suitable. And SDS method was also suitable to the extraction of dried C. sinensis. To investigate whether the hypoglycemic effect of C. sinensis, the inhibition of Cordyceps sinensis on the intestinal mucosal disaccharidase were studied by enzymatic means in vitro conditions, meanwhile, Panax Notoginseng, Dioscorea opposita were chosen as the control medicines. In the reaction system the final concentration of all the five herbs was 0.05g/mL, and the inhibition rate were 74.67%, 45.30%, 39.17%, 15.50% and 32.63%, respectively,while the inhibition rate of the positive control Acarbose was 72%. Whether the hypoglycemic effect of C.sinensis clinical application needs further study.