| Respiratory disease infected by is a susceptible and highly prevalent disease in avians.The mortality rate of the disease is 10%-30%,even 90% when the animal was infected with the strong virus strains.Liansang oral liquid is a formula with the functions of heat-clearing,detoxifying,relieving cough and pain,which is composed of Forsythiae Fructus,Mori Folium,Platycodonis Radix,Houttuyniae Herba,Glycyrrhizea Radix Et Rhizoma and other herbs.The study was to determine the efficacy and toxicity of Liansang oral liquid by establishing the quality control standard,pharmacodynamics and toxicology,which would provide scientific data for further clinical study and delevelop a new Chinese herbs formula for veterinary medicine.(1)Study on standard of quality contral of Liansang oral liquid.The thin-layer chromatography was used to detect the major elements in Forsythiae Fructus,Mori Folium,Platycodonis Radix,Houttuyniae Herba,Glycyrrhizea Radix Et Rhizoma respectively.Then high performance liquid chromatography was used to detect the forsythin,which is the major elements in Forsythiae Fructus.The results showed that similar spots were found in the same distance within the thin-layer chromatography plate when tested the Forsythiae Fructus,Mori Folium,Platycodonis Radix,Houttuyniae Herba,Glycyrrhizea Radix Et Rhizoma respectively and their contents in Liansang oral liquid,respectively.Diamonsil C18 column(250×4.6 mm,5 μm)was used to separate the elements from 10 μL of Liansang oral liquid under the condition: 1 m L/min of flow rate,30℃,and the consist of separative solution is acetonitrile:water=23/77 in volume.Forsythin in the Liansang oral liquid should be tested under ultraviolet wave in 277 nm.The above method meets the requirements of Chinese Veterinary Pharmacopoeia in specificity,precision,repeatability,stability,retrieve rate and durability,which is suitable for the determination of forsythia glycosides in Liansang oral liquid.(2)Pharmacodynamic study of Liansang oral liquid.The effects of Liansang oral liquid was investigated in Sprague-Dawley(SD)rats and Kunming(KM)mice,in which the effect heat-clearing was tested in fever SD rats model induced by yeast,the anti-inflommatory effects was comfirmed in paw swelling SD rats model induced by dextran40,000,the cough relieving effects was comfirmed in cough KM mice model induced by ammonia,the pain relieving effects was comfirmed in writhing KM mice model induced by acetic acid.The results showed that the body temperature of rats decreased highly significant in the medium(1.7 g/kg)and high dose groups(5.1 g/kg)at 6 h after modeling,compared with the model group(P<0.01).In the dextran 40,000-induced paw swelling test,the volume of the paw of rats from the medium(1.7 g/kg)and high dose groups(5.1 g/kg)decreased extremely significant at 4 h after modeling(P<0.01).The coughing and writhing times in mice were significantly reduced in the medium(3.8 g/kg)and high dose groups(11.4 g/kg)compared with the model group(P<0.01),indicating that Liansang oral liquid was able to clear heat,decrease the inflammatory response,relieve cough and pain.(3)Toxicological study of Liansang oral liquid.The acute and sub-chronic toxicity of Liansang oral liquid on KM mice and SD rats were studied.In Aacute toxicity testing: The 2male and 2 female KM mice was provided Liansang oral liquid at the dose of 20 m L/kg(1.94 g/m L)by intragastric administration for three times within 24 h.The toxicity of Liansang oral liquid in mice was observed and recorded for another 7 days.The results showed that no abnormal clinical manifestations and death was found on the tested mice.The maximum dosage study was tested on 40 KM mice,including 20 male and 20 female,which were randomly divided into two groups equally.The dose and method of administration were the same as mentioned above.The results showed that no abnormal behavior was observed in all the mice.Therefore,the maximum dose of Liansang oral liquid was 116.4 g/kg.The Sub-chronic toxicity was tested on SD rats,including control group,low(4.85 g/kg),medium(9.7 g/kg),and high dose group with 10 male and 10 female in each group.The tested rats were provided Liansang oral liquid for 30 days,and the clinical signs of the rats were observed and recorded.On the 31 st day of the study,all surviving rats were anesthetized and blood samples were collected for hematological and biochemical profile.The organs were collected and weighed to calculate organ index.And histopathological examination was performed on organs in the high-dose group.The results showed that no significant changes in clinical signs,hematology,blood biochemistry and organ coefficients caused by Liansang oral liquid were observed in the rats of all dose groups and tissues and organs.It indicates that there is no sub-chronic toxic effect of Liansang oral liquid 19.4 g/kg for longer time administrated. |
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