【作者】 葛金芳；

【导师】 李俊； 金涌；

【作者基本信息】 安徽医科大学 ， 药理学， 2004， 硕士

【摘要】 慢性支气管炎（慢支）是严重危害人类健康的常见病、多发病。目前认为，大气污染、吸烟、病原体感染及过敏因素等与慢性支气管炎的发病密切相关，但发病后的机体免疫功能状态直接和（或）间接影响慢性支气管炎的发生和发展。因此，纠正或调节机体免疫功能紊乱可能是防治慢性支气管炎的重要手段之一。 目前对慢性支气管炎的治疗主要着眼于抗炎、镇咳、平喘作用的研究，至今尚无令人十分满意的药物。而对于改善机体免疫功能状态的免疫调节药物，尤其是具有免疫调节作用的中药的研究还没有引起足够的重视。鉴于慢性支气管炎免疫功能紊乱的病理机制和TAL高效、低毒的作用特征，我们探讨了TAL对慢性支气管炎的治疗作用及其可能的作用机制。 本文在研究TAL抗炎及免疫调节作用的基础上，进一步观察了其镇咳、祛痰和平喘作用；同时采用正交设计实验方案，联合应用BCG和LPS构建了大鼠慢性支气管炎模型，探讨了TAL对慢性支气管炎的治疗作用及其作用机制。 具体研究内容包括4个部分： 1．TAL的抗炎及免疫调节作用 TAL(75，225，675mg·kg^-1)ig给药能明显减轻二甲苯诱导的小鼠耳肿胀和冰醋酸引起的小鼠腹腔毛细血管渗出；TAL(50，150，450mg·kg^-1)ig给药对角叉菜胶诱导的大鼠足爪肿胀、棉球诱导的大鼠肉芽肿和福氏完全佐剂诱导的大鼠佐剂性关节炎具有明显抑制作用。提示，TAL具有明显的抗炎作用。 TAL(75，225，675mg·kg^-1)ig给药能显著改善Cy诱导的免疫低下小鼠碳廓清指数（K值）和吞噬指数（a值）的降低；显著提高Cy诱导的免疫低下小鼠血清IgM和IgG的生成，增加Cy诱导的免疫低下小鼠脾细胞溶血素的产生；增强Cy安徽医科大学硕士学位论文引起的免疫功能低下小鼠的Dm反应;提示，TAL可调节Cy致免疫功能低下小鼠的非特异性免疫、体液免疫和细胞免疫。 体外实验证明，TAL（0.005，0.05，0.5，smg·L，）可明显增加conA、廿s诱导的AA大鼠脾淋巴细胞增殖反应和ILZ的产生，同时抑制AA大鼠PM。产生过高的IL-1:进一步提示，IAL具有较强的免疫调节作用。2.TAL的镇咳、祛痰、平喘作用 TAL（75，225，675 mg·ks-l）ig给药可延长氨水引起小鼠咳嗽的潜伏期、减少小鼠2分钟内咳嗽次数，并对小鼠气道酚红排泄有明显促进作用;TAL（45，135，405 mg·kg一‘）ig给药可延长拘榜酸引起的豚鼠咳嗽潜伏期并减少豚鼠5分钟内咳嗽次数;对磷酸组胺和乙酞胆碱混合液引起的豚鼠药物性喘息有保护作用，并可增加离体豚鼠支气管肺泡灌流量;提示TAL具有较强的镇咳、祛痰和平喘作用.3.慢性支气管炎（Chrenic Bronehitis，CB）动物模型的研究 尾静脉注射BCG(5nl助和气管内注入LPs(2oo。助联合应用，饲养3周后可出现较为典型的慢性支气管炎病理形态学改变，提示该方法是用于研究慢性支气管炎发生及发展机制的简便可靠的模型。4.IAL对慢性支气管炎的治疗作用及部分机制研究 TAL（50，150，450 mg·ks-l）ig给药可明显降低cB大鼠支气管肺泡灌洗液 （BA工F）中白细胞总数以及中性粒细胞和肺泡巨噬细胞比例的上升;可使CB大鼠支气管粘膜炎症浸润程度明显减轻;支气管腔内分泌物明显减少、支气管粘膜上皮细胞脱落明显减轻、可抑制支气管粘膜上皮细胞增生，使支气管内径增大，通气量增大。提示，TAL对慢性支气管炎的气道炎症有一定的改善作用。进一步研究发现，TAL（50，150，450 mg·kg一‘）ig给药可显著降低CB大鼠肺组织匀浆中升高的TNF-。、ILS，同时使下降的IL10水平恢复至接近正常水平，并对CB大鼠气道粘膜上皮细胞NF一KB、ICAM一1的过度表达具有一定的抑制作用。提示其对CB的治疗作用可能与其平衡细胞因子产生、调节机体免疫功能有关。更多还原

【Abstract】 Chronic bronchitis (CB) is a common airway disease of high incidence rate that threats human health. As regards to its etiology, air pollution, smoking, and airway infection have shown close relationships. But, the body immunological status may well play a more important role in the initiation and the progress of CB. Thus regulating the body immunological state is a critical role in the prevention and therapy of CB.Several studies of bronchial biopsy specimens and bronchoalveoar lavage fluid (BALF) from subjects with chronic bronchitis have provided evidences for a inflammatory and immunological impairment progress in the airway of these subjects, consisting predominantly of mononuclear cells and neutrophils. In these studies, cytokines such as tumor necrosis factor alpha (TNF- ), interleukin-8 (IL-8), and interleukin-6 (IL-6) are of particular interest. Meanwhile, cell adhesion molecules such as intercelluar adhesion molecule-1 (ICAM-1) and L-Selection, and Nuclear factor kappaB (NF-B) have also been shown close attention.Based on the theoretic research, we designed the experiments to observe the effects of Triterpene acids of Loquat. Leaf (TAL) on rat CB model which duplicated by combination of Bacillus Calmette Guein (BCG) and lipopolysaccharide (LPS) injected through caudal vein and endotracheal intubation respectively. The main contents were devided into some sections as follows: 1. Anti-inflammatory and immunoregulatory actions of TALTAL(75, 225, 675 mg -kg-1, ig) could significantly inhibit the mice ear edema, the rat palm swelling and the rat cotton pellet granuloma induced by dimethylbenzene,carregeenin, and cotton pellet wool respectively. The primary inflammatory and secondary inflammatory reaction (secondary swelling , multiple arthritis and pathologic change of ankle arthritis) reaction in adjuvant arthritis (AA) rats induced by Freund’s complete adjuvant (FCA) were also inhibited by TAL. The results showed TFS had a remarkable anti-inflammatory action.TAL(75, 225, 675mg kg-1, ig) apparently improved carbonic clearances of macrophages (PM ) and the hemolysin level in serum and splencyte in mice immunosuppressed by Cy. Delayed-type hypersensitivity (DTK), which was suppressed by cyclophosphamid (Cy), was enhanced by TAL(75, 225, 675mg kg-1, ig) in mice. These results suggested TAL could recover nonspecific, humoral and cellular immune function in mice with immunosuppression. Moreover, TAL(675mg kg-1, ig) significantly increased both CD4+ and CD8+ cell, especially influenced on CD4+ cell, which rendered the ascending ratio of CD4+ to CD8+ cell. These results suggested that the immunoregulatory actions of TAL might be related to its function of regulating T cell subgroup.TAL(0.005-50mg L-1) could not only increase ConA and LPS induced splenocytes proliferation and IL-2 production of splenocytes, but also reduce the elevated IL-1 production from peritoned macrophate (PM ) in AA rats in vitro.2. The antitussive, expectorant and antiasthmatic actions of TALTAL (75, 225, 675 mg kg-1, ig) could not only prolong the cough latent period and decrease the frequency of cough in mice, but also promote phenolsulfonphthalein excretion. Moreover, TAL (45, 135, 405mg kg-1, ig) could prolong the cough latent period and asthma incubation period, reduce the cough frequency, and increase the bronchoalveolar lavage amounts of guinea- pigs in vitro.The results showed TAL had strong antitussive, expectorant and anti-asthmatic actions.3. Establishment of chronic bronchitis rat model by combined utilization of BacillusCalmette-Guerin (BCG) and lipopolysaccharide (LPS)Through combining utilization of BCG (5mg, injected through caudal vein) and LPS (200 g, injected through endotracheal intubation), the typical morphologic changes of chronic bronchitis were induced after 3 weeks.The results showed that chronic bronchitis rat model established by combining utilization of BCG and LPS is convenient and simple for the longitudinal study on the mechanism of initiation and progress of chronic bronchit更多还原