| OBJECTIVE: Banlian Baidu Oral Liquid is composed of nine traditional Chinese medicines,Banlangen,Forsythia suspensa,Anemarrhena asphodeloides,Pogostemon patchouli,Acorus tatarinowii,Radix Rehmanniae,Gypsum,Rhizoma Rehmanniae and Tulip.The original name of Banlian Baidu Oral Liquid is Antiviral Oral Liquid,which is from the second supplement of the 1990 edition of the Pharmacop It can clear away heat and dampness,cool blood and detoxify.For wind-heat cold,fever,upper respiratory tract infection,influenza,mumps and other viral infections.The preliminary clinical application of veterinary medicine showed that the medicine had a remarkable effect in preventing and treating wind-heat cold in livestock and poultry.In order to develop it into a new veterinary drug,we have carried out preliminary experimental studies on its pharmacology,pharmacodynamics and toxicity.METHODS:(1)Study on the extraction technology and quality standard of Banlian Baidu Oral Liquid.Taking the content of Gaoyichun and Forsythia suspense glycoside and the solubility of Decoction and the extraction quantity of volatile oil as indexes,the water extraction process of Banlian Baidu Oral Liquid was optimized by orthogonal experiment;the content of Gaoyichun and Forsythia suspense glycoside was determined by HPLC,and the quality control standard of Banlian Baidu Oral Liquid was established by TLC identification of Radix Isatidis,Forsythia suspense,Anemarrhenae,Pogostemon and Acorus calamus.(2)Volatile oil experiment of nano-emulsion cladding plate Lianbaidu oral liquid.The best formulation of volatile oil encapsulated in nanoemulsion was evaluated and found by making pseudo ternary diagram,drug loading stability test and transmission electron microscopy observation.(3)Pharmacodynamics of Banlian Baidu Oral Liquid.Study on Antipyretic effect: Rabbits were injected with skimmed milk 1.3 g/kg through ear vein to make a fever model.The antipyretic effect of Banlian Baidu Oral Solution was investigated by observing the temperature rise and fall of mice after administration.Study on the effect of stopping cough: a cough model was made in mice by strong ammonia spray method.The mice were given medicine once a day before inducing cough.They were given continuous administration for one week and then concentrated with ammonia to induce cough.The cough reaction within 1 min was observed in mice.Anti-inflammatory effect study: The mice were induced by xylene to swell the auricle.The mice were given once a day for a week before the model was established.After the last Administration for 30 minutes,50 mu L xylene was applied on both sides of the right ear of each rat.After 45 minutes,the ear slices were killed and weighed to calculate the inhibition rate of inflammation.Study on bacteriostasis: Staphylococcus aureus was diluted 10 times in normal saline.In the series of dilution groups,0.5 m L bacterial solution was injected intraperitoneally into each rat to obtain the lowest total lethal concentration.KM mice were randomly divided into 5 groups.In addition to normal saline,the mice in other groups were given orally once a day for a week before infection.After 30 minutes of the last administration,the mice were injected intraperitoneally with the lowest lethal concentration of 0.5 ml,and the mice died within 3 days.(4)Study on safety pharmacology of Banlian Baidu Oral Liquid: Using MC4000 non-invasive blood pressure instrument and EMKA non-invasive pulmonary function instrument,the indexes of pulse,systolic blood pressure,diastolic blood pressure,tidal volume and respiratory frequency of mice in blank group and administration group were measured at 0,1,2 and 4 hours after administration,and the effects of drugs on cardiovascular and pulmonary function of mice were studied through pentobarbital sodium in mice.Suprathreshold and subthreshold dose sleep experiments were used to observe the effects of drugs on sleep time in mice.The number of spontaneous activities and dropping times in blank group and drug group were determined by mouse autonomic activity experiment and mouse functional coordination experiment.The effects of Banlian Baidu oral liquid on the nervous system of mice were studied.(5)Acute toxicity and long-term toxicity experiment of Banlian Baidu Oral Liquid: The highest dose group was 6000 mg/kg in acute toxicity experiment,the middle dose group was 1200 mg/kg,and the low dose group was 240 mg/kg.The number of mice died was recorded and 4/4 lethal dose and 0/4 lethal dose were calculated to find the LD50 dose of the drug;the long-term toxicity experiment was possible through target animals.The 30-day feeding experiment and 15-day normal feeding recovery experiment were conducted with 100-fold dosage of medicated feed.The blood routine,blood biochemistry,feed intake,water intake,body weight and organ coefficient of rats were measured on 30-day and 45-day respectively.(6)Study on Antipyretic and bacteriostatic mechanism of Banlian Baidu Oral Solution: The antipyretic pharmacodynamic experiment was carried out according to the pharmacodynamic part.The Antipyretic Mechanism of the drug was explored by measuring the indexes of c AMP,IL-1beta,IL-6 and PEG2 in rabbits.The bacteriostatic mechanism of volatile oil from the pathogen of Staphylococcus aureus was determined by measuring the minimum inhibitory concentration,growth curve,detection of bacterial soluble protein by polyacrylamide gel electrophoresis,detection of bacterial nucleic acid content by agarose gel electrophoresis,and detection of agr A and sar A virulence genes of Staphylococcus aureus by RT-PCR.RESULTS:(1)The optimum extraction technology of water decoction was as follows:(1)The ratio of medicine to liquid was 1:10,and the extraction time was 3 hours each time,and the extraction time was 3 times.The precision test results showed that RSD of Yichun was 2.7%,RSD of forsythiaside was 2.1%;the repeatability test showed that RSD of Yichun was 2.5%,RSD of forsythiaside was 2.2%;the average recovery of Yichun was 100.4%,RSD was 2.68%,the average recovery of forsythiaside was 102.73,RSD was 3.55%,and the negative control had no interference.The results of TLC showed that under the experimental conditions,the spots were clear,the negative control had no interference,the extraction efficiency was high and the reproducibility was good.(2)The experiment of inclusion of volatile oil by nano-emulsion method showed that the optimum formulation was EL-60,glycerol,vitamin E and water.The size of inclusion complex was mainly distributed at 80 nm,and the size range was below 100 nm.A uniform nano-emulsion was formed.(3)Antipyretic and anti-inflammatory tests showed that Banlian Baidu Oral Liquid could significantly reduce the body temperature of feverish rabbits and inhibit the degree of auricle swelling induced by xylene in mice.Antibacterial tests showed that the drug could reduce the mortality of mice infected with Staphylococcus aureus.Antitussive test showed that the drug could significantly increase the half cough time of mice induced by concentrated ammonia water,and the effective rate of high dose group was 160%.(4)Safety pharmacological experiments showed that there was no difference in pulse,systolic pressure,diastolic pressure,tidal volume,respiratory frequency between the administration group and the blank control group,and there was no difference in the number of spontaneous activities between the administration group and the pentobarbital sodium suprathreshold dose sleep time between the control group and the blank control group.The drug had no toxic side effects on the nervous system,respiratory system and cardiovascular system of the animal body.(5)Toxicity experiment showed that in acute toxicity experiment,the highest dose group of Banlian Baidu Oral Liquid was 6000 mg/kg,and there was no animal death.Therefore,LD50 was more than 5000 mg/kg,which was actually non-toxic.Long-term toxicity experiments showed that Banlian Baidu Oral Liquid had no toxic effects on growth curve,blood routine,blood biochemical indexes and visceral tissues of animals.(6)Experiments of antipyretic and antimicrobial mechanism showed that volatile oil could effectively inhibit the growth of Staphylococcus aureus,and could reduce the expression of soluble protein,total DNA,virulence factors arg A and sar A.Through transmission microscopy,the volatile oil of Banlian Baidu Oral Liquid could destroy the cell wall and cytoplasm of bacteria and lead to bacterial death.Through the above effects,bactericidal effect could be achieved.CONCLUSION: The extraction process of Banlian Baidu oral liquid and the inclusion process of its volatile oil were established.The content of the main effective components of Banlangen,Xiuyichun and Forsythia suspensa,were identified.The TLC identification of Banlangen,Forsythia suspensa,Acorus calamus,Anemarrhena asphodeloides and Rehmannia glutinosa were carried out,and the quality control standard of the drug was established.Pharmacodynamic studies have shown that Banlian Baidu Oral Liquid has good antipyretic,anti-inflammatory,bacteriostatic and antitussive effects.Its antipyretic effect is related to the reduction of c AMP,IL-1beta and PGE2.Its antimicrobial effect is related to the inhibition of bacterial growth,the destruction of bacterial structure,the inhibition of virulence gene expression and the reduction of bacterial DNA and soluble protein.Safety pharmacology and acute toxicity experiments have proved that Banlian Baidu oral is a safe drug with no toxic and side effects on three vital systems of animal body. |
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