| Dairy cows subclinical mastitis is one of the most common diseases with high incidence, whichcould cause serious economic losses in the dairy industry."Ru ning san" was made from many kinds ofChinese herbal medicines, such as Semen Vaccaria, Epimedium, Dandelion and other drugs, which isbased on the veterinary theory in veterinary prescription. It has the effect of anti-bacterial andanti-inflammatory, reducing swelling, pass through the milk. The paper focused on the partial study ofpreclinical evaluation of "Ru ning san", which makes basis for experimental and theoretical in clinicalapplications.1. To establish identification methods of Semen Vaccaria, Epimedium, Dandelion in "Ru ning san"by TLC. Results showed "Ru ning san", Control herbs and standard substance in the correspondingposition in the chromatogram displayed the same color spots, the corresponding blank-control had nopots. The TLC spots were clear, specificity, repeatability in the preparation, which can be used as "Runing san" quality control standards.2. To investigate the safety of "Ru ning san" for treatment or prevention of the mastitis in dairycow, the acute&sub-acute toxicity was tested in mice and rat. In acute toxicity test,60mice wererandomly divided into6groups, the animals were administered drug by intragastric infusion at singledose of7.83,9.21,10.84,12.75,15.00mg/kg and0.9%NaCl respectively. Poisoning typical symptomand death cases were observed and the median lethal dose (LD50) was calculated. In order to estimatethe maximum dosage,40mice were randomly divided into the treatment group and Control group,"Runing san" was administered intragastrically at a maximum concentration (1g/mL) and maximumcapacity (0.04mL/g) with single dose in the treatment group,0.9%saline was used in Control. Afterbeing administered, the mice were observed for7days. In sub-acute toxicity test,80rats were randomlydivided into3drug-treated Groups (high, middle and low dose) and one Control, drug was administeredorally at dose of3000,1500and750mg/kg, respectively. In Control, rats were administered0.9%NaCl. Body weight gain, routine blood test indexes, blood biochemical parameters, organ coefficientand histopathology were detected. The results showed there was not any poisoning and death cases ineach group, there was no LD50in this herbal compound. The maximum daily dose of "Ru ning san" wasover40.0g/kg in mice. These data indicated that the drug is actually non-toxin. There were notsignificantly different in body weight gain, routine blood test, blood biochemical indexes and organcoefficient between3drug-treated groups and Control group. The histopathological result was not anylesion in rat real organs of every group. All of these results suggested that "Ru ning san" was safety forthe dairy cow, which were in mastitis under the recommended dosage and course of the treatment inclinic.3. The results of clinical pharmacodynamics experiment showed that the efficient rate of "Ru ningsan" was88.89%, which is higher than the positive control medicine Wan Rukang (76.00%). Milk composition, blood physiological and biochemical indexes change of cows administered with "Ru ningsan" at various dosages showed no significant difference with those in control group (P＞0.05),indicating that "Ru ning san" possessed better efficacy for subclinical mastitis.4. We found that the concentrations of IL-8、TNF-α、MDA and the activities of T-AOC、SOD、GSH-Px、NAGase、MPO in samples of serum from cows were detected and compared by ELISA、visible spectrophotometry, which the cows were from the cured group and the uncured group. Theresults showed that "Ru ning san" have the effect of enhance the activities of T-AOC、SOD、GSH-Px,degrade the concentration of lipid peroxides MDA、inflammatory factors (IL-8, TNF-α), inhibit theactivities of MPO、NAGase in serum from cows with subclinical mastitis. |
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