| Cow endometritis is a kind of reproductive diseases with great harmfulness,which is caused by bacteria infection.Danlianhua uterine perfusate is composed of Coptidis rhizoma,Forsythiae fructus,Salvia miltiorrhizae radix et rhizoma and Carthami flos.It has the functions of clearing heat and dampness,promoting blood circulation and removing blood stasis.What’s more,it has a unique advantage and significant effect for the treatment of acute endometritis.Preclinical toxicology study is the premises and foundation of the safety and effectiveness of clinical medication.This study was conducted on the acute toxicology,chronic toxicity and target animal safety studies of Danlianhua uterine perfusate.All the results were showed below:(1)Acute toxicology test of Danlianhua uterine perfusateDetermination of the median lethal dose(LD50): Eight mice with half males and half females were divided randomly into two groups.By studying out the dose in a high dose of5000 mg/kg,it carried on the pre-experiment injecting Danlianhua uterine perfusate into mouth and abdominal cavity,respectively.Then each group added up to ten with double repeated experiments.The result showed that there was no death in the experiment.So,it confirmed that LD50 of Danlianhua uterine perfusate gavaged and intraperitoneal injected in mice are both greater than 5000 mg/kg.Maximum tolerance test: Forty mice with half males and half females were divided randomly into two groups.Then they were used to inject Danlianhua uterine perfusate into mouth by using the concentration of 2.33 g/mL in dose of 0.4 mL/10 g and abdominal cavity by using the concentration of 1 g/mL in dose of 0.2 mL/10 g,respectively.After continuously observed for 10 days,the mental state of mice,eating and drinking water,coat color and fecal character etc.were recorded.In the 7th day of experiment,the experimental mice were weighed and calculate the maximum multiples.The result showed that none of the mices gavaged and intraperitoneal injected with Danlianhua uterine perfusate died.And the MTDswere 93200 mg/kg and 20000 mg/kg,respectively,which convert into MTT was 466 and 100 times.Skin irritation test: Sixteen rabbits with half males and half females were divided randomly into four groups named complete skin groups(2 groups,single and multiple dose group)and damaged skin groups(2 groups,single and multiple dose group).In self contrast method,the influences of Danlianhua uterine perfusate on intact and damaged skin were investigated,respectively.Single dose groups were washed off the residual drugs with warm water after wiped Danlianhua uterine perfusate for 6 hours.And multiple dose groups were wiped once a day for seven consecutive days.After removing the residual drugs,the rabbits were observed by the erythema,edema and whether there was a roughness of the skin,pigmentation,bleeder or not.The results showed that the score of skin irritation reaction of each experimental rabbit was 0,which belonged to nonirritating substance.Mucous membrane irritation test: Six healthy female rabbits were randomly divided into experimental group and control group.Experimental group were slowly uterine perfused with2 mL using Danlianhua uterine perfusate.And control group were uterine perfused with the same amount of saline solution.After 24 hours,all the rabbits were put to death in method of gas bolt,and removed the complete vaginas which were opened longitudinally.Then top,middle and bottom of the vagina were macroscopic observed and histopathological exanimated that whether there was a congestion or edema.The results showed that the score of vaginal mucosa irritative reaction with perusing congestion and edema was 0.78,and the stimulus score and intensity of pathological tissue changes was 1.67 and 0.67,respectively,which belonged to nonirritating substance.(2)Chronic toxicity test of Danlianhua uterine perfusateEighty SD rats with half males and half females were divided randomly into four groups.The groups respectively were: high dose group of 20 g/kg(the original solution concentrated to 2 g/mL),middle dose group of 6.32 g/kg(the original solution diluted to 0.632 g/mL with5% twain-80),low dose group of 2 g/kg(the original solution diluted to 0.2 g/mL with 5%twain-80),and control group of 10 mL/kg solvent solution(5% twain-80).All rats were dosed 1 mL/100 g once a day for 30 consecutive days and were freedom to gather the food and drink.And the weight,feed intake,water quantity and clinical symptoms of rats were daily recorded.After the test finished,all rats were examined by histopathological method,detected the hematology and blood biochemical indicator,and calculated the viscera index of main organization.The results showed that the mental state,behavior,activities,coat andtraits of excrement and urine of experimental rats had no abnormal change during the dosing.The weight,levels of BWG,viscera indexes,hematology and blood biochemical indexes of experimental rats had no significant difference compared with control group in the same gender(P > 0.05).The viscera tissue morphologies of high dose group were not seen pathological changes that caused by drug toxicity.It shows that Danlianhua uterine perfusate is safe to be used in different doses for a long period.(3)Target animal safety testing of Danlianhua uterine perfusateTwenty healthy cows,randomly divided into 4 groups,respectively were 1 times,3 times and 5 times of the recommend clinical dose group and the blank control group.With the trial period for 7 d,each experimental group was respectively uterine perfusated 100 mL,300 mL and 500 m L Danlianhua uterine perfusate on the 2nd day,4th day and 6th day.Clinical examination,hematology,blood biochemical indexes and main histopathology were detected on the 1st day and 7th day.The results showed that the mental state,behavior,activities,coat and traits of excrement and urine of the experimental cows during the period of drug delivery had no abnormal change.Hematology and blood biochemical indexes,compared before and after the dosages,had no obvious difference(P > 0.05).The main viscera tissue morphologies of high dose group were not seen pathological changes.It shows that Danlianhua uterine perfusate can be safely applied on target animal. |
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