Research On Preparation Process,Quality Standard,Preliminary Safety And Pharmacodynamics Of Zhiqi Kangai Granules

The prescription of“Zhiqi Kangai granules”,an effective empirical prescription based on the selection and combination of“Buzhong Yiqi decoction”contained in Li Dongyuan,who was a medical doctor in the Jin and Yuan dynasties of China,was composed of eight kinds of traditional Chinese medicine,such as Astragalus membranaceus,ganoderma lucidum,Codonopsis pilossus,Scutellaria baicalensis,and so on.In this study,the preparation process,quality standards,preliminary safety and pharmacodynamics of Zhikang Granules were discussed,providing theoretical basis for production and clinical therapy.Methods:（1）The preparation technology of Zhiqi Kangai granules:The single-factor and orthogonal experimental design methods were used to determine the optimal water extraction and preparation process.（2）The quality standard of Zhiqi Kangai granules:The TLC and HPLC were used to research the qualitative and quantitative on active ingredients,which was determined the quality standard of the preparation.（3）The acute toxicity of Zhiqi Kangai granules:40 clean-grade Kunming mice were divided into the control group and the drug-administered group.While the drug-administered group was administered intragastrically a maximum drug concentration of 1 g/mL twice in 24 h with volume of 0.4 mL/10 g,the control group was given purified water.After i.g.for 14 days,the drinking water,body weight and organ index of the mice were determined,with the pathological sections of the main organs stained with HE,and the maximum tolerated dose in one day was calculated.（4）The pharmacodynamics research of Zhiqi Kangai granules:174 clean-grade Kunming mice were divided into the control group,the model group,Zhenqi Fuzheng granules group,and the low,medium,high dose group in Zhiqi kangai granules,with28 in each group.The immunosuppressive model was established by intraperitoneal injection（ip）of cyclophosphamide（CTX）in 80 mg/kg for 3 days,with boosted once every week.After i.g.for 14 days,the organ index,phagocytic phagocytosis,blood routine,T lymphocyte subsets,and the serum cytokines IL-6,TNF-α,IFN-γof the mice were detected,with HE tissue slices made.Results:（1）The preparation technology of Zhiqi Kangai granules:（1）The whole prescription medicine excluding Ganoderma lucidum were extracted 3 times,with 10,8,8 times of water,and 60,60,30 min of extraction time;Ganoderma lucidum was extracted 3 times,with 12,9,9 times of water,and 40 minutes each time.Then,the liquid medicine was concentrated to prepare the extract powder.（2）The preparation technology:The dry comprises were mixed with the soft starch at 1:1.5and moistened with 70%ethanol,then the soft materials were made,which were dried at 45℃to obtain the finished product.（2）The characteristic spots of Astragalus membranaceus,Scutellaria baicalensis,Ganoderma lucidum can be detected,with no interference in the negative control;The content of baicalin has a good linear relationship in the range of 0.212 to 2.120μg（r=0.9993）,with an average recovery of98.40%.（3）Compared with the control group,except for the slight degeneration of hepatocytes under the pathological section of the liver in the administration group,there were no obvious abnormal changes in other organs by macroscopic and microscopic examination;The maximum tolerated dose in mice was 217.2 times as much as that of clinical adults.（4）Compared with the control group,the thymus index in model group was decreased（p<0.01）,with the carbon clearance index K and the phagocytic indexαdeclined,and the microstructures of the thymus and spleen atrophied and disordered;The number of peripheral blood leukocytes,lymphocytes,red blood cells,and the content of hemoglobin were declined（p<0.05;p<0.01;p<0.01;p<0.01）,with the CD₈⁺%in peripheral blood,and the content of IL-6,TNF-α,IFN-γdeclined（p<0.01）,while CD₄⁺/CD₈⁺%increased（p<0.01）,indicating the success of the model.Compared with the model group,the number of white blood cells,lymphocytes in the medium dose group of Zhiqi kangai Granules were increased（p<0.05）;The thymus index in the high dose group was returned to normal,the thymus and spleen tissue structure were gradually recovered,with the clearance index K and the phagocytic indexαincreased（p<0.01）;While in the medium and high dose groups,the levels of CD₈⁺%,IL-6 and IFN-γwere increased（p<0.01;p<0.01;p<0.05）,the ratio at CD₄⁺/CD₈⁺was decreased（p<0.01）;In the high dose group,the levels of IL-6,TNF-αand IFN-γwere increased（p<0.01）.Conclusion:The optimized preparation process was obtained;The established quality control standard was stable and easy to operate;The quality control standard obtained by the test was stable and convenient to operate;The preparation has little toxicity,which was safe and reliable;The preparation could improve the non-specific and the specific immunity in immunosuppressed mice,thereby enhancing the immune function of the body.