【作者】 杨永贵；

【导师】 胡元亮； 戴鼎震；

【作者基本信息】 南京农业大学 ， 临床兽医学， 2004， 硕士

【摘要】 奶牛乳房炎的发病率高、危害大，而现行的治疗方法存在诸多弊端。在初步研制出中药银花乳炎清涂膜剂的基础上，本研究进一步观察了该药治疗奶牛乳房炎的疗效并探讨了其作用机理，具体工作分为以下几步： 1．药效试验 观察了银花乳炎清涂膜剂对奶牛临床型和隐性型乳房炎的疗效。结果表明，该药对重症临床型乳房炎的治愈率和有效率分别为73.33％和86.6％，青霉素对照组为80.0％和90.0％；对轻症临床型乳房炎的治愈率和有效率分别为87.8％和93.9％，青霉素对照组为83.3％和91.6％；对隐性乳房炎给药5天的治愈率和有效率分别为67.3％和85.6％，给药8天分别为83.6％和92.7％。提示该涂膜剂对临床型乳房炎疗效较好，尤其对隐性乳房炎疗效显著，但对重症病例最好配合其它药物。 2．安全性试验 以20倍临床用量涂布脱毛后的家兔皮肤，观察单次给药和多次给药(每天1次，连续7天)涂膜剂对完整皮肤和受损皮肤的刺激性反应。结果表明，除个别受损皮肤有轻微红斑外，其余均未见红斑和水肿。同时以150倍临床用量涂布家兔皮肤，并制备成煎剂大鼠灌胃给药，连续观察给药后7天内的急性毒性反应，结果均未见任何异常反应。说明该涂膜剂无刺激性和急性毒性。 3．抑菌试验 将乳炎清、方中单味药及其不同组合制成煎剂，用平板法观察各煎剂对无乳链球菌、停乳链球菌、金黄色葡萄球菌的体外抑制作用。结果表明乳炎清抑菌效果较好，金银花、紫花地丁和蒲公英起主要作用。同时测定了各煎剂对无乳链球菌乳酸脱氢酶(LDH)的影响，结果表明，金银花、蛇床子、金银花-紫花地丁-蒲公英组合和蛇床子-紫花地丁-蒲公英组合在0.0625g／mL时能够降低LDH活力。 4．抗炎试验 用小鼠耳廓肿胀法和大鼠足跖肿胀法观察银花乳炎清煎剂的抗炎作用，结果表明乳炎清煎剂能够明显降低二甲苯所致的小鼠耳廓肿胀及10％鸡蛋清溶液所致的大鼠足跖肿胀，具有显著的抗炎作用。 5．透皮试验 用水提醇沉法从乳炎清主药金银花中提取醇溶物，按乳炎清涂膜剂的制法加入基质材料，制成涂膜剂，加到装置在改良Franz扩散池的奶牛乳房皮肤上，每小时取样1次，连续12h，用高效液相色谱法测定涂膜剂内绿原酸的透过量。结果表明，绿原酸以零级动力学经皮渗透，提示透皮给药方法是可行的。能够维持较为恒定的血药浓度。更多还原

【Abstract】 Dairy cattle mastitis is a disease with high incidence rate, which could lead to great loss. There are many shortcomings in present therapeutic methods. Based on primary development of Yinghua-ruyanqing (RYQ) balm, its curative effects and mechanism for dairy cattle mastitis were studied in present experiments. The research included five sections as follows.1. Curative experiment The curative effect of RYQ balm was observed for clinical and subclinical dairy cattle mastitis in this experiment. The results showed that cure rate and effective rate of the balm was respectively 73.3% and 86.6% for serious clinical mastitis, while penicillin, 80.0% and 90.0%. For mild clinical mastitis, the cure rate and effective rate was 87.8% and 93.9%, while penicillin, 83.3% and 91.6%. After treating for 5 days, the cure rate and effective rate was 67.3% and 85.6% for the subclinical one, for 8 days, 83.6% and 92.7%. Those suggested that RYQ balm possessed better efficacy for clinical and subclinical mastitis, and it was necessary to cooperate with other drugs in serious case.2. Safety experiment First, the back skin of rabbits were denuded and smeared with the RYQ balm for one and seven times at 20 times clinical dosage after one side was damaged, and then the stimulation and acute toxicity to skin of the drug were observed. The results indicated that there were no erythema and edema except for the damaged skin in one rabbit. Second, the back skin of rabbits was smeared with the RYQ balm and rats were poured into stomach once with RYQ decoction at 150 times clinical dosage. The acute toxicity reactions in the skin and whole body were observed for seven days. The results showed that there were no any abnormal reactions. Those suggested that the balm were safety.3. Antibacterial experiment The RYQ, its composed medicines and their composition were prepared into decoctions whose inhibitions to Streptococcus agalactiae, Streptococcus dysgalatiae and Staphylococcus aureus were observed by plating method.The results showed that antibacterial effect of RYQ was better, in which honeysuckle flower, tokyo violet and mongolian played main role. At the same time, effects of these decoctions on LDH of Streptococcus agalactiae were determined. The determination confirmed that honeysuckle flower, common cnidium fruit and two compositions (one was honeysuckle flower, tokyo violet and Mongolian; another was common cnidium fruit -tokyo violet-mongolian) could decrease the activity of LDH at 0.0625 g/mL.4. Anti-inflammatory experiment The anti-inflammation effects of RYQ decoction were tested by mice auricle-swelling and rat metatarsus-swelling method. The results showed that the decoction could obviously reduce the auricle swell caused by dimethylbenzene and the metatarsus swell caused by 10% solution of egg white, which confirmed that RYQ had better anti-inflammation effect.5. Skin-penetrating experiment Crude Chlorogenic acid was extracted from Honeysuckle flower, main constituent of RYQ, by water-extraction and ethanol precipitation method and was made into balm. The skin-penetrating amount of Chlorogenic Acid in modified Franz-diffusion device was determined each hour within 12 hours by means of HPLC. The results showed that the permeating-skin process of Chlorogenic Acid accorded with zero-order kinetics, which suggested that skin-penetrating administration was available and could maintain invariable concentration in blood.更多还原