Preparation Of Huoxuevinao Formula Granule And Anti-Cerebral Ischemia Effects Of Hirsutella Sinensis Mycelium

Limited cerebral ischemia causes cerebral blood supply disorders and induces the hypoxia. Ischemic stroke is charactered as a high rate of incidence, morbidity and mortality diseases. In the paper, the preparation of Huoxueyinao formula granule and the anti-cerebral ischemia effects of Hirsutella sinensis mycelium are studied. Huoxueyinao formula granule was composed of the extractions of Puerariae Lobatae Radix, Scutellariae Radix, Morinda officinalis Radix, and et al. Puerariae Lobatae Radix,Scutellariae Radix, and et al were extracted together with ethanol, and Morinda officinalis Radix was extracted with water. Orthogonal design was utilized to optimize the extracting conditions of ethanol extracting group and water extracting group respectively. The evaluation indexes are puerarin and baicalin for ethanol extracting group and nystose for water extracting group in orthogonal tests. The amount of puerarin, baicalin and nystose were determined by HPLC. The optimum extraction conditions were to extract three times using tenfold 60% ethanol and 1.5h per time for ethanol extracting group, and to extract three times using eightfold water and 1.5h per time for water extracting group. The extraction powder of Huoxueyinao formula was obtained by spray drying and mixed with excipients including dextrin and lactose. Huoxueyinao Formula Granule was obtained by dry granulation method, which met the quality requirements of 2010 Chinese Pharmacopoeia.The middle cerebral artery occlusion (MCAO) models of ICR mouse and SD rat were used to study anti-cerebral ischemia effects of Hirsutella sinensis mycelium. The results of neurobehavioral test and triphenyltetrazolium chloride (TTC) staining of experimental animal indicated that Hirsutella sinensis mycelium was able to ameliorate neurobehavioral behavior and decrease the infarct percentage significantly. Caspase-3 expression level in the core and penumbra tissue was measured using western blotting and immunohistochemistry and the results showed that the caspase-3 expression level in the penumbra tissue of sham group and treatment group was significantly higher than the vehicle group but the caspase-3 expression level in the core tissue was nearly equivalent among the above three groups.