Development And Pharmacodynamic Study Of Jinqing Granule Of Pig Gastric Ulcer

Pig gastric ulcer is a high frequency disease,which induced by many factors,can caused a big economic loss.Due to the complexity of its causes,the pathogenic mechanism is unknow,there is no effective treatment of this disease.The goal of this subject is to observe the pharmacological effects of "Jinqing Granule" on three differents gastric ulcer models and one gastritis model,and research on its effective mechanism.The main experiment content and results are as follows:1.Bacterial culture and identification:Collecting clinical fresh pig stomach tissue for bacterial culture.Using morphology observation,biochemical identification and 16s rDNA gene identification to identified the bacterial species.Results:A strain of Helicobacter pylori was effectively isolated from pig stomach.2.Drug screening and establishing formula:Using ethanol reflux,water decoction or ultrasonic method,extracts of 25 different traditional antipyretic and antidote medicines were obtained.Inhibition zone diameters of above 25 extracts on Helicobacter pylori from swine were detected by Oxford cup method.Minimal inhibitory concentration(MIC)were detected by macrodilution broth method,and combined antibacterial effects in vitro of them were also observed.We determined the MICs of the extracts obtained from Coptidis rhizoma,Canarii fructus and Lonicerae japonicae flos against Hp ranged from 1.56 to 6.25 mg·mL-1.The MICs of the extracts obtained from Isatidis folium,Scrophulariae radix,Sophorae ronkinensis radix et rhizome,Pulsatiliae radix,Violae herba,Rhei radix et rhizome and Taraxaci herba ranged from 12.50 to 25.00 mg·mL-1.The MICs of the extracts obtained from Sargentodoxae caulis,Polygoni cuspidate rhizome et radix,Belamcandae rhizome,Lonicerae japonicae caulis,Chrysanthemi flos,Houttuyniae herba,Forsythiae fructus,Isatidis radix,Leonuri herba,Chebulae fructus and Ampelopsis radix ranged from 50.00 to 100.00 mg·mL-1.The MICs of the extracts obtained from Cimicifugae rhizome,Dryoteridis crassirhizomatis rhizome and Bupleuri radix ranged over 200.00 mg·mL-1.The results of pairwise joint bacteriostatic test showed that the fractional inhibitory concentration index(FICI)of the extracts against Hp,Canarii fructus and Tinosporae Radix was less than or equal to 0.5.The FICI of Canarii fructus and Coptidis rhizome,Tinosporae Radix and Coptidis rhizome were more than 0.5,but less than or equal to 1.The FICI of Canarii fructus and Lonicerae japonicae flos,Tinosporae Radix and Lonicerae japonicae flos,Coptidis rhizome and Lonicerae japonicae flos were more than 2.It was concluded that Canarii fructus,Tinosporae Radix,Coptidis rhizome and Lonicerae japonicae flos showed a good antibacterial effect against Hp in vitro.The combination of Canarii fructus and Tinosporae Radix was characterized by a synergism effect,whereas the combination of Canarii fructus,Tinosporae Radix and Coptidis rhizome was characterized by a additive effect.The combination of Lonicerae japonicae flos and any other medicine showed a antagonism effect.Results:Tinosporae Radix and Coptidis rhizome were chosen to be drug formulas,and named this drug "Jinqing Granule".3.The optimization of extraction and preparation technology of Jinqing Granule:(1)Using the orthogonal experimental method to optimization of extracting conditons.Using ethanol as the solvent,and using ethanol concentration,solid-liquid ratio,extraction time,extraction times as the examine factors,choose L9(34)orthogonal table to do this experiment.Results:the optimum extraction processe of Tinosporae Radix is 10 times of ethanol,extract second times each time lasts for 30min.The optimum extraction processe of Coptidis rhizome is 8 times of ethanol,extract three times each time lasts of 1.5h.(2)According to the results of single factor experiment,using the granular molding rate,moisture absorption,bulk density,melting time as examining index,observing the therapeutic effects of granulating when using different proportions of cane sugar,dextrin,lactose and glucose as supplementary material,to select the best preparation technology of Jinqing Granule.Results:Sucrose and dextrin were chosen to be the auxiliary materials of Jinqing Granule,and the ratio of drug and excipients for 1:3.Granules by this method is conform to the law of the People's Republic of China pharmacopoeia 2015 edition,and not hygroscopic,combined with the actual production costs,then determine this method to be the preparation technology of Jinqing Granule.4.Establishing quality standard of Jinqing Granule:On the basis of "Veterinary drug research technical guidelines assembly(2006-2011)",estabilishing the quality standard for Jinqing Granule,to make sure it conform the national drug quality standard requirements.Results:A scientific and feasible quality standard of Jinqing Granules was estabilished.5.The pharmacodynamics study of Jinqing Granule:Establishing three differents gastric ulcer models and one gastritis model to observe the efficacy of Jinqing Granule.Results:Jinqing Granule has a good prevention and treatment effect for different gastric ulcer models and gastritis model.Jinqing Granule can resistant the gastric damage,anti-oxidation,anti-inflammatory and anti-bacterial effect.The main mechanism of Jinqing Granules are reducing TLR-4 and NF-? B p65's expression in the gastric mucosa and controlling the ratio of Bcl-2 and Bax.In conclusion,Jinqing Granule have a good prevention and therapeutic effect on three different gastric ulcer model and a gastritis model by helicobacter pylori strain from pig.In addition,Jinqing Granule have process stability,reliable quality and a good development potential.