Comparative Study On Preparation Process And Efficacy In Formula Granules,Compatible Granules And Traditional Decoctions Of Gypsum-Anemarrhenae

Objective:Xiangxu compatible medicines are widely used in TCM clinic.This topic choose Gypsum and Anemarrhenae,two representative traditi-onal Chinese medicine,in order to investigate that whether granules have the same compatibility effect as decoctions by comparative study on eff-icacy and content of active ingredient in formula granules,compatible granules andtraditional decoctions of Gypsum-Anemarrh Enae.This rese-archer can not only provide methodological reference for the comparative study of different preparations of various compatible Chinese medicine,but also provide experimental basis for the clinical application of traditi-onal Chinese medicine granules with the same compatibility relationship.Methods:1.Orthogonal test was used to optimize the preparation technology of Gypsum formula granules,Anemarrhenae asphodeloides formula granules and gypsum-Anemarrhena asphodeloides compatible granules.Main technological parameters were evaluated by the content of Ca²⁺and mangiferin and the extraction rate.The selection and amount of the best excipients,the concentration and amount of wetting agent in the forming process were evaluated by the indexes of particle forming rate,fluidity and moisture absorption rate.The critical relative humidity and water content of particles were gauged.2.Comparison of content of active ingredient:EDTA titration was used to detect the content of Ca²⁺,and high performance liquid chromatography was used to detect the content of mangiferin content in each preparation.3.Comparison of antipyretic efficacy:A total of 80 healthy adult SD male rat ranging from 200 to 220g were selected and randomly divided into experimental groups according in line with their weights.8 in each group:Negative control group,model group,positive control group（aspirin）,Gypsum decoction group,Gypsum formula granule group,Anemarrh Enae decoction group,Anemarrh Enae formula granule group,Gypsum-Anemarrh Ena decoction group,Gypsum-Anemarrh Enae compatible granule group,Gypsum-Anemarrh Ena formula granule group.The fever model of rats was established by subcutaneous injection of20%dry yeast suspension（10ml/kg）.Four hours after the establishment,the rat fever model（ΔT>0.8℃）was successfully established,and the corresponding liquid was given to each group by gavage.The Anal temperature of rats was measured every 1 hour after the establishment of the model for 10 times.The changes of body temperature in each group were recorded and the maximum temperature rise height（ΔTmax）was calculated.The contents of IL-6,TNF-αand PGE2 in the hypothalamus were detected by ELISA.4.Comparison of efficacy of eliminating hot in stomach:A total of80 healthy adult SD male mice ranging from 200 to 220g were selected and randomly divided into experimental groups according in line with their weights.8 in each group:negative control group,model group,positive control group（Zuo Jin Wan）,Gypsum decoction group,Gypsum formula granule group,Anemarrh Enae decoction group,Anemarrh Enae formula granule group,Gypsum-Anemarrh Enae decoction group.Gypsum-Anemarrh Enae compatible granule group,Gypsum-Anemarrhe-nae formula granule group.The rat model of gastric heat syndrome was established by intragastric administration of ginger water decoction（5g/kg）and capsaicin ethanol mixture（10ml/kg）.After 15 days,the drug group was given the corresponding solution once a day,and the negative control group and the model group were given the same amount of saline by gavage for 10 days.The body temperature,diet and water consumption of rats in each group were recorded daily.The pathological changes of gastric tissue were observed by HE staining,and the contents of IL-2,IL-1βand IL-10 in serum were detected by ELISA.Results:1.The optimum extraction technology of the Gypsum is adding12 times of water,extract 3 times,60 minites each time.The best molding process is dextrin was used as the auxiliary material and 70%ethanol was used as the wetting agent,dextrin accounted for 10%of the main drug,and the wetting agent accounted for 50%of the main drug.The optimum extraction technology of Anemarrhena asphodeloides formula granules is adding 12 times of water,extracting three times for 30 minutes each time,and forming technology is extract-soluble starch（1:1.2）,granulating with85%ethanol as wetting agent.The optimum extraction technology of gypsum-Anemarrhena asphodeloides Compatible Granules is adding 12times of water,extracting twice,60 minutes each time,and forming technology is extract-soluble starch（1:1）,granulating with 85%ethanol as wetting agent.The moulding rate,fluidity,solubility,moisture absorption rate and moisture content of the three kinds of particles all meet the standards of granules.2.Comparison of content of active ingredient:（1）Comparison of Ca²⁺:Compared with Gypsum decoction,the content of Ca²⁺in Gypsum formula granules,Gypsum-Anemarrhenae compatible granules and Gypsum-Anemarrhenae formula granules decreased,but there was no significant difference（P>0.05）;There was no difference in Ca²⁺content among the three granules.The content of Ca²⁺in Gypsum-Anemarrhenae decoction and Gypsum decoction was not significantly different.（2）Comparison of Mangiferin:Compared with Anemarrhenae decoction,the content of mangiferin in Anemarrhena formula granules,Gypsum-Anemarrhenae compatible granules and Gypsum-Anemarrhenae formula granules were decreased（P<0.05）.There was no difference in mangiferin content among the three granules,and the content of mangiferin in Gypsum-Anemarrhenae decoction and Anemarrhenae decoction was similar.3.Comparison of antipyretic efficacy:（1）Temperature changes:4 hours after modeling,the body temperature of the model group continued to rise,compared with the negative control group,there was a statistical difference（P<0.05）,indicating that the rat fever model was successful;1-3 hours after administration,the body temperature of the rats in each group remained steadily rising,and the temperature difference of the aspirin group decreased faster than that of the model group（P<0.05）;4-5 hours after administration,there was a statistical difference between the aspirin group and the model group（P<0.05）.Compared with the model group,the body temperature of rats in aspirin group returned to normal,with significant difference（P<0.01,P<0.05）,but there was no significant difference among other groups（P>0.05）.6 hours after administration,the body temperature of rats in each group maintained a steady downward trend.The body Temperature of aspirin group,Anemarrhenae decoction group,Gypsum-Anemarrhenae decoction group and Gypsum-Anemarrhe-nae compatible granules group was all significantly lower than that in model group（P<0.05）.（2）Maximum rise body temperature（ΔTmax）:After modeling,theΔTmax of the model group increased significantly compared with the negative control group（P<0.01）;compared with the model group,theΔTmax of aspirin group decreased significantly（P<0.05）,while theΔTmax of other groups did not decrease significantly.（3）The expression level of IL-6、TNF-αand PGE2:The levels of serum IL-6,TNF-αand hypothalamic PGE2 in the model group were higher than those in the negative control group 6 hours after administration（P<0.01,P<0.05,P<0.05）;compared with the model group,the levels of IL-6,TNF-αand PGE2 in the other group decreased（P<0.05 or P<0.01）.4.Comparison of efficacy of eliminating hot in stomach:（1）Change of rat weight:There was no significant difference in the weight of each group compared with that of the negative control group at the early stage of the model establishment;at the late stage of the model establishment,the weight of the negative control group continued to rise,while the weight of the model group increased slowly and with a small extent（P<0.05）.After administration,the rat weight of each group increased rapidly and tended to be stable,and there was no significant difference compared with the negative control group.（2）Change of rat’s food intake:There was no significant difference in rats’food intake in each group（P>0.05）in the early 7th stage of model establishment;in the late 8th stage of the model establishment,the rats’food intake in the model group decreased gradually compared with the negative control group,with significant difference（P<0.01）;After administration,the rats’food intake in each group increased compared with the day of model establishment,and increased compared with the model group,too.（3）Change of water intake in rats:There was no significant difference in water intake among groups in the early 7th stage of model establishment（P>0.05）.In the late 8th stage of the model establishment,the water intake of rats in model group increased gradually compared with that of negative control group（P<0.01）.After administration,the water intake of rats in each group decreased compared with the day of model establishment,and decreased compared with that of model group.（4）Changes of rat feces and behavior of gastric perfusion:in the late stage of modeling,the feces of the model group were indurated than those of the negative control group;after administration,the hardness of the feces decreased;in the model group,rats experienced severe struggle,hoarse,writhing and other phenomena,which made it more difficult to feed;after administration,the above behaviors were ameliorated.（5）Pathological changes of gastric tissues:The gastric mucosal defect,exfoliation and erosion were observed in the model group.Mucosal lamina propria was infiltrated by more inflammatory cells,and the infiltration of inflammatory cells exceeded the bottom of the mucosal layer.In the administration group,the mucosal defect was weakened,the mucosal lamina propria was less infiltrated by inflammatory cells,and the infiltration of inflammatory cells was limited to the bottom of the mucosal layer.The pathological condition was improved compared with the model group.（6）The expression level of IL-2、IL-1βand IL-10:Compared with model group,the contents of IL-2 and IL-1βin serum decreased significantly（P<0.05）,while the contents of IL-10 increased significantly（P<0.05）after compatible use of Gypsum and Anemarrhena.There was a downward or upward trend in other groups.Conclusion:（1）The preparation process of three kinds of granules is reliable and stable,which can provide reference for practical production.（2）Chemical constituents:The contents of calcium ion and mangiferin in the three granules are slightly lower than those in the decoction,but there is no significant difference.（3）Pharmacodynamic comparison:Analysis from dosage forms,whether used alone or in combination,the antipyretic effects of decoction and granule are not significantly different;Analysis from before and after the compatibility,whether in decoction or granule,the antipyretic effects of compatible use is better than that of single.In TCM clinic,for gypsum and Anemarrhenae,its formula granules can be used instead of decoction,and compatible granules can be popularized in TCM clinic.