Study On The Preparation Technics And Main Pharmacodynamics Of Lingqi Granule

Objective: To research on the preparation process and quality control of Lingqi Granules and the function of Lingqi Granules in regulating the immune function of mice with immune hypofunction,gastrointestinal motility of mice with spleen deficiency and effects of digestive enzymes,for providing the experimental basis for the clinical application of Lingqi Granules.Methods: Orthogonal experiments were carried out to optimimze the extraction process and taking extraction rates of astragaloside and salvianolic acid B as investigation indexes.Taking granule sizes,hygroscopicity,shaping rates and dissolubility as investigation indexes,single-factor experiments were conducted to screen types and doses of excipient and determine the optimum preparation process of Lingqi Granules.The HPLC method was applied to determine the contents of astragaloside and salvianolic acid B in Lingqi Formula and the TLC method was adopted to conduct qualitative identification of Lingzhi,tanshinone IIA and icariin.Cyclophosphamide was utilized to duplicate the mouse model of immune hypofunction and intragastric administration of Huangqi granules(1.5g·kg-1)and Lingqi Granules(amounting to 9,4.5 and 2.3g·kg-1 of crude drugs)was conducted.Cytophagy,Tcell subsets and their cytokines,NK cells were used as indicators to evaluate effects of Lingqi Granules on immune function of mice.The mouse model of bonemarrow haematopoietic hypofunction caused by cyclophosphamide and 60 Co irradiation was used to observe influences of Lingqi Granules on blood routine examinations of mice.The mouse model of spleen deficiency resulted from Dahuang and reserpinwas applied to observe effects of Lingqi Granules on resistance against anoxia and intestinal motility of mice.Results: The optimum extraction process of Lingqi Granules was M2N3P2Q2.which were extracted three times by 10 times 50% ethanol one hour per time.The best excipient was dextrin with a ratio of dry extract to excipient of 1:1.25 and the corrigent was sucrose with a ratio of dry extract to corrigent of 1:3.Verification results of three batches of the preparation process showed that this preparation process was stable,feasible,reasonable and reliable with stable parameters of obtained granules,according with the standard of granules in “Chinese Pharmacopoeia”.The determination method of contents of astragaloside and salvianolic acid B was established,the satisfactory precision,stability,repeatability and sample recovery rate of the method meet the requirements.Contents of astragaloside and salvianolic acid B in Lingqi Granules were 0.0699% and 0.3891% respectively.The qualitative identify-cation of Lingzhi,tanshinone IIA and icariin meet the requirements of “Chinese Pharmacopoeia”.Intraperitoneal injection of cyclophosphamide could lead to immune hypofunction of mice.Lingqi Granules could increase leukocyte and lymphocyte counts in peripheral blood of mice,strengthen transformation of Con A-induced splenic T lymphocyte,activity of NK cells and enhance the carbon clearance index,the phagocytosis index,the spleen index,the thymus index,Value HC50 and the number of hemolysis plaque of mice.In the model group,CD3+ of mice increased,Th1/Th2 reduced and Th17 cells,Tregcells,IL-17 and IL-23 increased.In groups of Hunagqi and Lingqi Granules,CD3+ of mice reduced,Th1/Th2 increased and Th17 cells,Tregcells,IL-17 and IL-23 reduced.Cyclophosphamide and 60 Co irradiation could cause reduction of peripheral hemogram of mice significantly,while Lingqi Granules could enhance it(WBC,RBC and Hb)significantly.The resistance against anoxia of mice with spleen deficiency reduced and their intestinal peristalses accelerated.Lingqi Granules could significantly improve the resistance against anoxia of mice with spleen deficiency and slow down their intestinal peristalses.Conclusion: The complete formula of Lingqi Granules were extracted three times by 10 times 50% ethanol one hour per time.The preparation process was reasonable and reliable.The qualitative identification of Lingzhi,tanshinone IIA and icariin and contents of astragaloside and salvianolic acid B were used as quality standards which leads to controllable product quality.Lingqi Granules could enhance cellular and humoral immunity of mice,reduce CD3+,increase the ratio of Th1/Th2 cells and improve peripheral hemogram and symptoms of spleen deficiency.