苦参止痢胶囊对犬细小病毒病的治疗作用

发布时间：2019-06-09 21:06

【摘要】：犬细小病毒病因其高传染性和高死亡率,成为危害犬最大的疫病之一。由于用于辅助治疗的各种化学药物会带来耐药性等一系列问题,使得研发新型高效、无毒无副作用的抗犬细小病毒病中药制剂成为新课题。苦参止痢胶囊是根据中兽医理论和卫气营血辨证施治的原则,选用苦参、白芍、木香等组成的中药复方胶囊制剂,具有清热燥湿,凉血止痢的功效。本文通过细胞试验、动物试验和临床用药来探讨苦参止痢胶囊对犬细小病毒病的治疗作用。试验一高效液相色谱法测定苦参止痢胶囊有效成分的含量采用反向高效液相色谱法测定苦参止痢胶囊中苦参碱、氧化苦参碱、芍药苷、去氢木香内酯和木香烃内酯的含量。采用Phenemonex C18(250mm×4.6mm,5μm)色谱柱,苦参碱和氧化苦参碱用10mmol/L的醋酸胺水溶液(用氨水调节pH8)-甲醇梯度洗脱的流动相,紫外检测波长为210nm,流速:1mL/min;芍药苷用乙腈-0.1%磷酸溶液(14:86)的流动相,紫外检测波长为230nm,流速:1mL/min;去氢木香内酯和木香烃内酯用甲醇-水(65:35)的流动相,紫外检测波长为225nm,流速:0.65mL/min。结果苦参止痢胶囊中苦参碱为2.2mg/g、氧化苦参碱为1.05mg/g、芍药苷为9.1ng/g和去氢木香内酯为13.5mg/g,木香烃内酯由于含量过低未测出。试验二苦参止痢胶囊在细胞上对犬细小病毒的作用研究为了观察苦参止痢胶囊体外对犬细小病毒的作用,以FK8]细胞为载体,通过间接免疫荧光试验(IFA),用荧光素标记进入细胞的犬细小病毒,在荧光倒置显微镜下观察,用ImageJ软件分析荧光。结果显示先加中药再感染病毒组和中药与病毒作用后感染组都能显著减少FK81细胞中犬细小病毒的表达量,先加中药再感染病毒组平均减少量为36%,中药与病毒作用后感染组平均减少量为46%;先感染病毒再加中药组FK81细胞中病毒表达量与阳性组无显著差异。表明苦参止痢胶囊溶液在细胞上能有效阻止犬细小病毒的吸附和侵入,对于在细胞内复制的犬细小病毒无明显抑制作用。试验三中西药治疗犬细小病毒病的临床试验为了观察苦参止痢胶囊对犬细小病毒病的治疗作用,用水平感染的7只病犬作为动物模型,分为中药组、西药组和阳性组,分别为3只、3只和1只,1只健康犬为空白,用苦参止痢胶囊和西药治疗,通过临床症状、病理解剖和组织病理学切片光镜及电镜观察。结果显示:苦参止痢胶囊和西药都能控制犬细小病毒病,其中苦参止痢胶囊组没有出现血痢的症状,组织病理学切片观察肠道组织也没有明显的出血,并且维持了肠绒毛结构的完整性,表明苦参止痢胶囊能抑制犬细小病毒引起的肠黏膜损伤和出血性病变,从而对犬细小病毒病达到治愈作用。试验四苦参止痢胶囊治疗犬细小病毒病的扩大临床试验为了进一步探讨苦参止痢胶囊防治犬细小病毒病的效果。方法:利用动物医院门诊收治的犬细小病毒病病例,采用纯西药治疗与中西药联合治疗的对比试验,统计两种药的治愈率、治愈疗程等情况。结果.:用纯西药治疗的32只犬细小病毒病病例的治愈率为75%;应用苦参止痢胶囊的中西药联合治疗的32只犬细小病毒病病例的治愈率为87.5%;且中西药联合治疗的平均治愈疗程为5.32d,低于纯西药治疗的平均治愈疗6.29d。中西药联合治疗组治愈犬平均止住血痢天数为3.86d,显著低于西药组的5.38d(P0.05)。结论:苦参止痢胶囊对犬细小病毒病有良好辅助治疗作用。
[Abstract]:The cause of canine parvovirus is its high infectivity and high mortality, which is one of the most dangerous diseases of the dog. The novel high-efficiency, non-toxic and side-effect anti-canine parvovirus disease traditional Chinese medicine preparation has become a new problem due to a series of problems such as drug resistance and the like caused by various chemical drugs used for adjuvant treatment. Radix Sophorae Flavescentis anti-dysentery capsule is prepared from Radix Sophorae Flavescentis, Radix Paeoniae Alba, and Radix Aucklandiae, and has the effects of clearing away heat, eliminating dampness, and cold blood and stopping dysentery, and is prepared from Radix Sophorae Flavescentis, Radix Paeoniae Alba, and Radix Aucklandiae. In this paper, the therapeutic effect of Radix Sophorae Flavescentis on the canine parvovirus disease was discussed by cell test, animal test and clinical medication. A high performance liquid chromatography (HPLC) was used to determine the content of matrine, oxymatrine, Paeonia lactiflora, dehydrocostunolide and costunolide in the capsule of Radix Sophorae Flavescentis with high performance liquid chromatography. The mobile phase of a 10 mmol/ L aqueous solution of acetic acid (pH 8)-methanol gradient was used with a Phenemonex C18 (250 mm, 4.6 mm,5. mu.m) column, and a mobile phase with a wavelength of 210 nm and a flow rate of 1 mL/ min with an ultraviolet detection wavelength of 210 nm and a flow rate of 1 mL/ min; and a mobile phase of a paeonol-0.1% phosphoric acid solution (14:86), The ultraviolet detection wavelength is 230 nm, the flow rate is 1 mL/ min, the mobile phase of the dehydro-costunolide and the costunolide with methanol-water (65:35), the ultraviolet detection wavelength is 225nm, the flow rate is 0.65 mL/ min. Results The matrine was 2.2 mg/ g, the oxymatrine was 1.05 mg/ g, the peony root was 9.1 ng/ g and the dehydro-costunolide was 13.5 mg/ g, and the content of the costunolide was not measured. in ord to observe that effect of the radix sophorae flavescentis anti-dysentery capsule on the canine parvovirus in the cell, in order to observe the effect of the radix sophorae flavescentis anti-dysentery capsule on the canine parvovirus in vitro, the FK8] cell is taken as a carrier, and the canine parvovirus which enters the cell is labeled with a fluorescein by an indirect immunofluorescence test (IFA), The fluorescence was analyzed with ImageJ software under a fluorescence inversion microscope. The results showed that the expression of the canine parvovirus in the FK81 cells was significantly reduced by the combination of the traditional Chinese medicine reinfected group and the infection group of the traditional Chinese medicine and the virus, the average reduction of the first Chinese medicine reinfected virus group was 36%, and the average reduction of the infection group after the effect of the traditional Chinese medicine and the virus was 46%; No significant difference was found between the expression of the virus and the positive group of the virus plus the group of FK81 in the traditional Chinese medicine group. It is shown that the solution of the anti-dysentery capsule of the radix sophorae flavescentis can effectively prevent the adsorption and the invasion of the canine parvovirus, and has no obvious inhibiting effect on the canine parvovirus which is reproduced in the cell. In order to observe the therapeutic effect of Radix Sophorae Flavescentis anti-dysentery capsule on canine parvovirus disease, the three-and-western medicine in the treatment of canine parvovirus disease was used as animal model, and it was divided into three groups: the traditional Chinese medicine group, the western medicine group and the positive group. 3,3 and 1,1 healthy dog were blank, and were treated with flavescent sophora root, antipullorum and western medicine, and were observed by light and electron microscope with clinical symptoms, pathological anatomy and tissue pathology. The results showed that both the anti-dysentery capsule and the western medicine of the radix sophorae flavescentis can control the canine parvovirus disease, wherein the radix sophorae flavescentis antidiarrheal capsule group does not have the symptoms of the blood pullorum, the pathological sections of the tissue do not have obvious bleeding in the intestinal tissue, and the integrity of the intestinal villi structure is maintained, It is shown that the anti-diarrhea capsule of the radix sophorae flavescentis can inhibit the intestinal mucosa injury and the hemorrhagic lesion caused by the canine parvovirus, thereby achieving the cure effect on the canine parvovirus disease. In order to further study the effect of Radix Sophorae Flavescentis for preventing and treating canine parvovirus disease, the clinical trial was conducted to study the effect of Radix Sophorae Flavescentis for preventing and treating canine parvovirus disease. Methods: In the case of canine parvovirus disease treated by the outpatient department of the animal hospital, the comparison between the treatment of pure western medicine and the combination of the Chinese and Western medicine was adopted, and the cure rate and the course of treatment of the two kinds of drugs were counted. As a result. the cure rate of 32 canine parvovirus disease cases treated with pure western medicine is 75%, the cure rate of 32 canine parvovirus disease cases treated with the combination of the Chinese and western medicines of the radix sophorae flavescentis anti-diarrhea capsule is 87.5%, and the average curing course of the combination therapy of the Chinese and western medicines is 5.32d, The mean cure was 6.29 d, which was lower than that of pure western medicine. The average number of patients cured by the combination of Chinese and Western medicine was 3.86d, which was significantly lower than that of the western medicine group (P0.05). Conclusion: The anti-dysentery capsule of Radix Sophorae Flavescentis has good adjuvant treatment effect on canine parvovirus disease.
【学位授予单位】：扬州大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S858.292

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