卡泊芬净雾化吸入防治侵袭性肺曲霉病药效学与安全性的动物实验研究

发布时间：2019-03-22 13:16

【摘要】：侵袭性肺曲霉病(invasive pulmonary aspergillosis, IPA)多发生于重度免疫缺陷人群,死亡率高。抗真菌药通过雾化吸入给药近来受到广泛关注。与系统用药相比,雾化给药少量药物即可达到肺部高药物浓度,并且因较少入血降低了药物系统毒性。本研究的目的是建立免疫抑制小鼠IPA模型,并评价雾化吸入卡泊芬净在预防和治疗IPA的疗效及安全性。主要分为以下三个部分： 1.免疫抑制小鼠侵袭性肺曲霉病模型的建立目的：动物实验被广泛的应用于侵袭性肺曲霉病(IPA)诊断及治疗方面的研究,本文利用气管插管法建立免疫抑制小鼠IPA模型。方法：以环磷酰胺及地塞米松对小鼠进行免疫抑制处理后,气管插管气管内滴入烟曲霉孢子悬液,通过组织病理进行验证,并观察小鼠生存-时间曲线。同时运用平板菌落计数方法比较气管插管法与传统滴鼻法造模实际进入鼠肺的孢子量。结果：气管插管法可成功建立小鼠IPA模型。小鼠1周内死亡率55%,接种24小时后肺组织匀浆平板菌落计数显示气管插管法所造成的孢子入肺量为5.17±0.32CFU(1g)/g肺组织,而传统滴鼻法仅为3.82±0.49CFU(1g)/g肺组织。结论：气管插管法成功建立小鼠IPA模型,较滴鼻法更适于进行IPA相关研究。 2.卡泊芬净雾化吸入给药防治侵袭性肺曲霉病药效学研究目的：通过动物实验对雾化吸入卡泊芬净对IPA的防治效果做初步探讨。方法：免疫抑制小鼠IPA模型,分别给予卡泊芬净腹腔注射及雾化用药,用药策略分为单纯治疗及预防+治疗,同时设置标准治疗——两性霉素B腹腔注射组及对照组,持续用药至感染后第7天。比较各组小鼠生存率及肺组织真菌负荷。结果：与标准治疗——两性霉素B腹腔注射相比,雾化吸入卡泊芬净作为单纯治疗性用药、或预防+治疗用药均能有效降低小鼠肺组织真菌负荷；但只有作为预防+治疗给药时才能有效提高小鼠生存率,且对IPA预防效果与腹注给药相当。结论：动物实验证明卡泊芬净雾化吸入给药是一种有效的预防IPA的给药方式。 3.卡泊芬净雾化吸入给药局部安全性研究目的：观察连续多次雾化吸入卡泊芬净后小鼠肺组织病理学及局部炎症因子表达。方法：ICR小鼠分为三组,高剂量组每天雾化2小时,低剂量组每天雾化时间为1小时,设置生理盐水对照组,各组小鼠连续用药7天。各组小鼠行肺组织病理学检查并予以评分；提取肺组织RNA行荧光定量PCR,检测各组小鼠肺组织炎症因子表达。结果：雾化给药结束后各组肺组织病理评分无明显差异,荧光定量PCR检测显示雾化吸入卡泊芬净并未导致肺组织IL-1β、IL-6、TNF-α等炎症因子的活化。结论：动物实验证明卡泊芬净雾化吸入给药对肺组织刺激性小,安全性较好。综上所述,抗真菌药物雾化吸入用药在肺组织局部形成较高药物浓度,作为预防性用药时可显著增强抗真菌效果,且避免了高血药浓度导致的副作用。本研究利用动物实验观察了雾化卡泊芬净用于预防和治疗IPA疗效和安全性,与标准治疗药物——两性霉素B治疗相比,雾化卡泊芬净作为预防性用药时可提高小鼠生存率,并降低肺真菌负荷。雾化用药易于操作且安全性较好,有潜在的临床应用前景。
[Abstract]:Invasive pulmonary aspergillosis (IPA) mostly occurred in the severe immunodeficiency population, and the mortality was high. Antifungal agents have recently received extensive attention through the use of aerosolized inhalation. Compared with the system, the drug can reach the high drug concentration of the lung, and the toxicity of the drug system is reduced due to the less entry of blood. The purpose of this study was to establish an IPA model for immunosuppression mice and to evaluate the efficacy and safety of nebulization in the prevention and treatment of ipa. It is mainly divided into the following three parts: Sub-point:1. The model of invasive pulmonary aspergillosis in immunosuppressed mice Objective: The animal experiment was widely used in the diagnosis and treatment of invasive pulmonary aspergillosis (IPA). Method: After the immunosuppression treatment of mice with cyclophosphamide and dexamethasone, the spore suspension of Aspergillus fumigatus was dropped into the trachea of the tracheal intubation, and the mice were observed by tissue pathology and the mice were observed. Survival-time curve. The method of plate colony counting is used to compare the actual intake of the tracheal intubation and the traditional nasal drop method. Spore volume of the rat lung. Results: The tracheal intubation can be successful The mouse IPA model was established. The death rate was 55% in the first week of the mouse, and the count of the lung tissue homogenate plate after 24 hours of inoculation showed that the amount of the spore entering the lung was 5.17 and 0.32 CFU (1 g)/ g of the lung tissue, whereas the conventional nasal drop method was only 3.82 and 0.49 CFU. (1 g)/ g lung tissue. Conclusion: The tracheal intubation method successfully established the mouse IPA model. suitable for IPA-related studies.2. Capofungin Pharmacodynamic Study on the Prevention and Treatment of Invasive Pulmonary Aspergillosis: by the Animal Experiment A preliminary study was made on the control and control of ipa. Methods: The model of IPA in immunosuppression mice was given. The administration strategies were divided into simple treatment and prevention + treatment, and the standard treatment _ amphotericin B was injected into the abdominal cavity. And the control group, and the drug was continuously applied to the 7th day after the infection. The results showed that, compared with the standard treatment _ amphotericin B, the effect of nebulization on the pulmonary tissue fungal load of the mice was effectively reduced compared with that of the standard treatment _ amphotericin B. Can be used for preventing and treating the administration of the mouse, and the mouse life can be effectively improved. Conclusion: The experimental results of the animal experiment prove that the Capofene is pure and fog. The administration of inhalation is an effective administration of the prevention of IPA Methods.3. The purpose of the local safety study of the administration of the aerosol inhalation of the caspofungin: to observe the continuous multiple fog Methods: ICR mice were divided into three groups, the high-dose group was nebulized for 2 hours a day, and the low-dose group was atomized daily. The time is 1 hour, and the normal saline control group is set, and the mice in each group are continuously used for 7 days. The lung tissue pathology of each group of mice is examined and scored, and the lung tissue is extracted The expression of inflammatory factors in lung tissue of each group was detected by fluorescence quantitative PCR of RNA. Results: There was no significant difference in the pathological score of lung tissue after the end of the drug administration. So as to lead to the activation of the inflammatory factors such as IL-1, IL-6, TNF-1, and the like in the lung tissue. In conclusion, the drug of the anti-fungal drug is used for the local formation of higher drug concentration in the lung tissue. In order to prevent and treat the effect and safety of ippofungin in the prevention and treatment of ipa, the effect and safety of the aerosolized caspofungin in the prevention and treatment of ipa were observed, and the treatment with the standard therapeutic drug _ amphotericin B was observed. Compared with the method, the survival rate of the mice can be improved when the atomized caspofungin is used as a preventive medicine, and
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R519

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