| Objective This study is undertaken to evaluate the pharmacodynamic mechanism of atomization inhalation of the traditional Chinese medicine Lemahui injection.Methods Experiments were carried out through the in vitro antibacterial tests and in vivo studies.(1)The antimicrobial tests in vitro:The determination of minimum inhibitory concentration(MIC)with trace broth dilution method to Lemahui injection on 6 kinds of common acute lower respiratory tract infection of pathogenic bacteria(Staphylococcus aureus,Streptococcus pneumoniae,Escherichia coli,Group A-?hemolyticstreptococcus,Klebsiella pneumoniae,Pseudomonas aeruginosa)standard strains and clinical strains of bacteria in different concentration of liquid,to study the inhibitory effect of Lemahui injection on pathogenic bacteria of common acute lower respiratory tract infection.(2)Animal experiment:By establishing the rat dry yeast induced fever model,mouse xylene induced ear swelling model,mouse ammonia induced cough model,mouse tracheal phenol red excretion model,to study the effect of atomized inhalation of lemahui injection on the anal temperature of rats,the ear swelling of mice,the cough latency and the number of coughs of mice,and the effect on the phenol red excretion of mice tracheal and determine the pharmacodynamic effect of atomized inhalation with Lemahui injection.(3)At the same time as pharmacodynamics research on dry yeast induced fever model rats and ammonia water induced cough model mice,we used ELISA technology to detect the expression of cytokines in animals to explore the mechanism of drug action.We used the indexes of rat serum IL-6,TNF-?,IL-1?,the content of PGE2 in hypothalamus and the content of IL-10 in mice serum as the observation target to explore the antipyretic and antitussive mechanism of Lemahui injection.Research results1.Lemahui injection showed different degrees of bacteriostatic effect on the standard and clinical strains of Staphylococcus aureus and Pseudomonas aeruginosa.The MIC of Staphylococcus aureus was 281.25-562.5ug/ml,that of Pseudomonas aeruginosa was 281.25-1125?g/ml,and the MIC value increased with the increase of the concentration of the suspension.The MIC values of Staphylococcus aureus and Pseudomonas aeruginosa clinical strains and standard strains were similar,and the two results were equal or one dilution difference.2.Lemahui injection showed no obvious bacteriostatic effect on Streptococcus pneumoniae,Escherichia coli,Group A-?hemolyticstreptococcus and Klebsiella pneumoniae.3.By subcutaneous injection of dry yeast suspension on the back,the rat model of fever can be successfully replicated.The heat potential of the model has the typical hypothermia stage,rise stage and peak plateau stage,and reaches the maximum temperature rise range at 7h.The anal temperature of rats was measured every hour after modeling and the temperature change was observed for 10 hours.In the third and fourth hours,compared with the model group(atomized normal saline),the anal temperature difference in the medium dose group of Lemahui injection decreased significantly(P<0.05),and the fever inhibition rate reached 197.56%in the fourth hour.However,there was no significant difference in TRI10(P>0.05).4.The content of 1L-1?,1L-6 and TNF-?n serum and the content of PGE2 in hypothalamus of yeast induced fever rats were detected.? The content of 1L-1(3 in the serum of the rats in the high,medium and low dose groups of inhaled atomized Lemahui injection was lower than that in the model group,but the difference was not statistically significant(P>0.05).? The content of IL-6 and TNF-? in the serum of rats in the medium dose group of inhaled atomized Lemahui injection was significantly lower than that in the model group(P<0.05).? There was no significant difference in the content of PGE2 in the hypothalamus between inhaled atomized Lemahui injection group and the model group(P<0.05).5.In the ear swelling test,the ear swelling degree of the high dose group of inhaled atomized Lemahui injection was significantly lower than that of the model group(atomized normal saline,P<0.01),and the inhibition rate of ear swelling was 29.35%.The ear swelling degree of the medium and low dose groups of inhaled atomized Lemahui injection was lower than that of the model group,but the difference was not statistically significant(P<0.05).6.In terms of reducing the number of coughs:?The number of coughs in the high,medium and low doses of Lemahui injection was less than that in the model group 1(atomized normal saline),and there were significant differences between the high and low doses of Lemahui injection and the model group 1(P<0.05),The results showed that high and low doses of inhaled atomized Lemahui injection could significantly reduce the number of coughs in mice caused by ammonia.?There was a significant difference in the number of coughs between the positive drug group(Dextromethorphan Hydrobromide oral solution gavage)and the model group 2(normal saline gavage,P<0.01).The number of coughs in the high dose and low dose groups of inhaled atomized Lemahui injection was higher than that in the positive drug group.The drug activity was 76.97%and 77.27%respectively,but there was no significant difference among the three groups(P>0.05).The results showed that the effect of atomized inhalation of medium and low doses of Lemahui injection was similar to that of Dextromethorphan Hydrobromide gavage.7.In the aspect of prolonging cough incubation period:?Model group.1 and model group 2 were treated with atomized saline and saline gavage respectively,and the cough incubation period of atomized saline was significantly longer than that of saline gavage(P<0.01);positive drug group(Dextromethorphan Hydrobromide oral solution gavage)was significantly different from model group 2(gavage saline)(P<0.01),but there was no significant difference with model group 1(P>0.05).It shows that atomization inhalation has a good antitussive effect.?The cough latency of the high dose group of inhaled atomized Lemahui injection was significantly different from that of the model group 1(atomized normal saline,P<0.01)and that of the positive drug group(P<0.01).The drug activity was 137.72%of the positive drug.The results showed that the antitussive effect of high dose inhaled atomized Lemahui injection was better than that of Dextromethorphan Hydrobromide gavage.?There was no significant difference between the medium,low dose group of inhaled atomized Lemahui injection with the positive drug group(P>0.05).The results showed that the antitussive effect of medium and low doses of atomized inhalation Lemahui injection was similar to that of Dextromethorphan Hydrobromide gavage.8.The level of IL-10 in the serum of the high,medium and low dose group of inhaled atomized Lemahui injection was significantly higher than that of the model group 1(atomized normal saline,P<0.01,P<0.05,P<0.01),and there was no significant difference between these groups and the positive drug group(P>0.05).There was no significant difference between the three dose groups(P>0.05).The results showed that the three doses of inhaled atomized Lemahui injection could significantly increase the expression of IL-10 in the serum of ammonia induced cough mice.9.The results of phenol red excretion test in mice showed that the phenol red excretion in the high and medium dose groups of inhaled atomized Lemahui injection was significantly higher than that in the model group(atomized aerosol injection,P<0.05,P<0.01),and there was no significant difference between the two groups with the positive drug group(P>0.05).There was no significant difference in phenol red excretion between the low dose group and the model group.It is suggested that atomization inhalation of high and medium doses of Lemahui injection has a good expectorant effect.Conclusion1.Lemahui injection can inhibit Staphylococcus aureus,Pseudomonas aeruginosa standard strains and clinical strains to varying degrees.The inhibition is affected by the amount of bacteria inoculated.Lemahui injection had no significant inhibitory effect on Streptococcus pneumoniae,Escherichia coli,Group A-?hemolyticstreptococcus and Klebsiella pneumoniae.2.The antipyretic effect of atomized inhalation of Lemahui injection may be related to the decrease of IL-6 and TNF-ain serum of febrile rats.3.Atomization inhalation of Lemahui injection has a good anti-inflammatory effect,can inhibit the ear swelling of mice caused by xylene,and reduce the acute inflammatory response.4.Atomization inhalation of Lemahui injection has a significant antitussive effect.It has a good effect on reducing the number of coughs and prolonging the latent period of coughs in mice induced by ammonia.The antitussive effect of the high dose group is better than that of the positive drug Dextromethorphan Hydrobromide oral solution gavage.The mechanism of antitussive action may be related to the increase of serum IL-10 level.5.Atomization inhalation of Lemahui injection has a good expectorant effect and can increase the amount of phenol red excretion in the trachea of mice. |
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