椒目油对哮喘豚鼠白细胞介素4,5及气道反应性的影响
作者： 赵睿，戚好文，谢柏梅，李志超。
【Abstract】 AIM: To explore the effects of Jiaomu Oil on interleukin4,5 in serum and airway hyperresponsiveness (AHR) of asthmatic model in guinea pig and to provide theoretical basis for treatment of asthma using Jiaomu Oil . METHODS: This experiment included 4 groups: normal control group, asthmatic model group, dexamethasone treatment group and Jiaomu Oil treatment group, involving 10 guinea pigs in each group. Asthmatic models in guinea pigs were constructed by immunization of intraperitoneal injection with saline ovalbumin (OVA) and challenge of inspiration with nebulized OVA. A threeway cannula was inserted into the lumen of the trachea for mechanical ventilation and the intratracheal pressure (IP) was measured simultaneously for reflecting airway responsiveness. The levels of IL4,5 in serum of guinea pigs were determined using enzymelinked immunosorbent assay (ELISA). RESULTS: The levels of IL4,5 of asthmatic model group［(38.2±3.4) and (344.4±21.8) ng/L］ were significantly higher than those of normal control group［(19.8±1.2) and (77.8±26.0) ng/L］(P0.01). The levels of IL4,5 of Jiaomu Oil［(22.8±3.5) and (89.2±38.7) ng/L］and dexamethasone groups［(22.0±1.8) and (234.6±35.1) ng/L］were both significantly lower than those of asthmatic model group(P0.01). The level of IL4 of Jiaomu Oil group didnt have any significant difference from that of dexamethasone group(P0.05), While the level of IL5 of Jiaomu Oil group was significantly lower than that of dexamethasone group(P0.01). The airway responsiveness of asthmatic model group ［(0.013±0.014) g/L］ was significantly higher than that of normal control group ［(0.168±0.186) g/L, P0.01］. Compared with that of asthmatic model group, the airway responsiveness of Jiaomu Oil ［(0.160±0.180) g/L］ and dexamethasone group ［(0.144±0.154) g/L］ both decreased significantly (P0.01), but they didnt have any statistical significance from that of normal control group (P0.05). The airway responsiveness of Jiaomu Oil group didnt have any statistical significance from that of dexamethasone group (P0.05). CONCLUSION: Jiaomu Oil could efficiently decrease the AHR of asthmatic guinea pig. The mechanism may be that Jiaomu Oil could prevent the production of IL4,5 in asthmatic guinea pig.
【Keywords】 Jiaomu Oil; asthma; interleukin4; interleukin5; airway hyperreactivity。
【摘要】目的： 探讨椒目油对哮喘豚鼠模型白细胞介素4,5(IL4,5)水平及气道高反应性(AHR)的影响，为椒目油治疗哮喘提供理论依据. 方法： 实验分为4组： 正常对照组、哮喘模型组、地塞米松治疗组和椒目油治疗组，每组各10只豚鼠. 用卵白蛋白(OVA)腹腔注射致敏和雾化吸入激发建立哮喘豚鼠模型，三通管做气管插管进行机械通气，测定气道内压力(IP)以反映气道反应性，ELISA法检测豚鼠血清中IL4,5含量. 结果： 哮喘模型组豚鼠血清IL4,5含量［分别为(38.2±3.4)和(344.4±21.8) ng/L］显著高于正常对照组［分别为(19.8±1.2)和(77.8±26.0) ng/L］(P0.01)；椒目油组IL4,5含量［分别为(22.8±3.5)和(89.2±38.7) ng/L］和地塞米松组IL4,5含量［分别为(22.0±1.8)和(234.6±35.1) ng/L］均明显低于哮喘模型组(P0.01)；椒目油组豚鼠IL4含量与地塞米松组比较无统计学意义(P0.05)，但IL5含量显著低于地塞米松组(P0.01). 哮喘豚鼠模型组气道反应性(0.013±0.014 g/L)与正常对照组(0.168±0.186 g/L)比较显著升高(P0.01)；椒目油治疗组与地塞米松治疗组气道反应性［分别为(0.160±0.180)和(0.144±0.154) g/L］与哮喘模型组比较均显著降低(P0.01)，与正常对照组比较无统计学意义(P0.05)；椒目油组气道反应性与地塞米松组相比无显著性差异(P0.05). 结论： 椒目油能有效降低哮喘豚鼠AHR，其机制可能是抑制哮喘豚鼠体内IL4,5的生成.
【关键词】 椒目油；哮喘；白细胞介素4；白细胞介素5；气道高反应性。
支气管哮喘是一种变态反应性疾病，IL4,5在其发病机制中占重要地位［1］. 目前临床上治疗哮喘多以对症为主. 中药椒目治疗哮喘历史悠久，疗效显著，但对其止哮平喘的作用机制还不清楚. 我们旨在通过探究椒目油对哮喘豚鼠血清IL4,5含量及气道高反应性(Hyperresponsiveness, AHR)的影响，揭示椒目治疗哮喘的作用机制，为椒目油运用于临床提供理论依据.
1材料和方法。
1.1材料健康雄性豚鼠40只，体质量(250±50) g（第四军医大学实验动物中心）；椒目油(第四军医大学西京医院药剂科，批号： 20050221)；卵白蛋白(OVA, Sigma公司)；磷酸组胺(上海丽珠东风生物技术有限公司)；戊巴比妥钠(国药集团化学试剂有限公司，批号： F20041117)；豚鼠IL4,5 ELISA试剂盒［美国Repidbio(RB)公司］；401AI超声雾化器(上海鱼跃医疗设备有限公司)；动物呼吸机DH140B(浙江医科大学医学仪器实验厂)；PTM9医用微机实控仪(二道生理仪)，RM6280多道智能生理信号记录系统(第四军医大学病理生理学教研室).
1.2方法。
1.2.1豚鼠支气管哮喘模型的建立参照文献［1］. 每日激发豚鼠1次，共7 d. 每次激发直到豚鼠出现咳嗽、呼吸急促、腹肌紧张等哮喘症状时为止.
1.2.2实验分组及处理用随机数字表法将豚鼠分为4组： 正常对照组、哮喘模型组、地塞米松治疗组和椒目油治疗组，每组各10只. 正常对照组用等量生理盐水致敏和雾化激发. 哮喘模型组用OVA致敏激发. 地塞米松治疗组致敏激发方法同哮喘模型组，但激发后立即给予5 g/L地塞米松溶液雾化吸入5 min. 椒目油组致敏方法同哮喘模型组，激发后立即给予椒目油和熟精油的混合物(按1∶1混合)，以3 mL/kg灌胃.
1.2.3气道反应性的测定参照文献［2］，以气道内压力(IP)上升至基线值的20%时所对应的磷酸组胺浓度(PC20)做为体现气道反应性高低的指标.
1.2.4测定IL4,5含量气道反应性测定完毕后，从豚鼠心脏直接抽血5 mL，静置30 min后以2000 r/min离心. 收集上层血清，运用ELISA法测定血清IL4,5的含量，具体步骤参照试剂盒说明书.
统计学处理： 所有数据均以x±s表示. 将原始数据输入SPSS 11.0统计软件包，气道反应性的组间比较运用KruskalWallis H法和Nemenyi法检验；IL4,5含量的组间比较运用SNKq（StudentNewmanKeulsq）法检验. 显著性水平均为0.05.