摘要
目的观察孕鼠在孕期补充不同浓度1α,25-二羟维生素D3后，相应仔鼠经卵蛋白致敏与激发后变应性气道炎症的差异，探讨孕期孕鼠1α,25-二羟维生素D3（VitD3）水平对子代幼鼠气道变应性气道炎症的影响。方法30只8周龄BALB/c小鼠按雌雄比例2∶1分笼饲养，采用随机数字表法分为5组：空白组，哮喘组，50、100、150 ng 1α,25-二羟维生素D3组（50、100、150 ng组），每组6只。阴栓出现表明受孕，记为D0。50、100、150 ng组孕鼠从D0起隔日于皮下注射相应剂量1α,25-二羟维生素D3 0.1 mL。空白组、哮喘组孕鼠从受孕当天隔日于皮下注射生理盐水0.1 mL。孕鼠产仔后，从各组雌仔鼠中随机选取6只致敏/激发。仔鼠5周龄时哮喘组、50 ng组、100 ng组、150 ng组分别于D0和D2腹腔注射卵清白蛋白（ovalbumin，OVA）0.05 mg＋氢氧化铝5 μg（0.5 mL），D7～12将致敏小鼠置于密闭容器中，用1%的OVA溶液15 mL超声雾化激发，每天1次，每次30 min。末次激发后24 h处理各组仔鼠，采用HE染色法观察肺组织病理变化，单盲法检测支气管肺泡灌洗液（bronchoalveolar lavage fluid，BALF）中细胞总数及分类，ELISA法检测BALF中IL-4、IL-5、IFN-γ水平和血清IgE水平。结果50、100、150 ng组仔鼠变应性气道炎症较哮喘组明显减轻。与哮喘组相比，50、100、150 ng组仔鼠BALF中细胞总数和嗜酸性粒细胞（eosinophil，EOS）均明显减少（P〈0.05），且随VitD3剂量增加而减少，呈剂量依赖性，各组间差异有统计学意义（P〈0.05）；BALF中IL-4、IL-5水平明显下降（P〈0.05），且随VitD3剂量增加而减少，呈剂量依赖性，各组间差异有统计学意义（P〈0.05）；而IFN-γ水平明显增高，且随VitD3剂量增加而增加，呈剂量依赖性，各组间差异有统计学意义（P〈0.05）；血清中IgE水平明显下降（P〈0.05），且随VitD3剂量增加而减少，呈剂量依赖性，各组间差异有统计学意义（P〈0.05）。结论孕鼠补充VitD3可以明显减轻仔鼠变应性气道炎症，该作用呈剂量依赖性。
Objective To investigate the effects of 1α,25-dihydroxyvitamin D3 （VitD3） supplement during pregnancy on allergic airway inflammation of its offspring in a mouse allergic asthma model. Methods Thirty eight-week-old BALB/c mice were kept according to gender proportion （female/male： 2/1）, and were randomly divided into 5 groups （6 mice per group）： control, asthma group, 50 ng VitD3 group （50 ng group）, 100 ng VitD3 group （100 ng group）, and 150 ng VitD3 group （150 ng group）. Observing the fertilization condition of female mice, day zero （D0） was recorded when vaginal plug was found. The pregnant mice in the 50 ng group, the 100 ng group and the 150 ng group were treated with subcutaneous injection of 0.1 mL VitD3 of corresponding dose from D0 to D20 （every other day）. The pregnant mice in the control and the asthma group were treated in the same way but with 0.1 mL saline. The female offspring mice were selected to continue the study, 6 in each group. The asthma group, the 50 ng group, the 100 ng group and the 150 ng group mice were treated by intra-peritoneal injection of ovalbumin （OVA, 0.05 mg） and aluminum hydroxide （5 μg） as an adjuvant on D0 and D2 in order to establish the mouse allergic asthma model. These sensitized mice were kept in a closed container （25 cm×12 cm×12 cm）, and challenged with 1% OVA 15 mL 30 min per day from D7 to D12. For analysis, all mice were sacrificed in 24 h after the last challenge. Histopathological analysis was performed on each mouse per group with HE staining. The number of cells and their classification in bronchoalveolar lavage fluid （BALF） were assayed. Levels of IFN-γ, IL-4, and IL-5 in BALF as well as serum immunoglobulin E （IgE） were measured by ELISA. Results According to the pathological sections of lung, the inflammation conditions of the 50 ng group, the 100 ng group and the 150 ng group were eased as compared to the asthma group. Comparing with the asthma group, both total cells and the percentage of eosinophile （EOS） were decreased in BALF in the 50 ng group, the 100 ng group and the 150 ng group, showing statistical difference from those in the asthma group （P〈0.05） and negative dose dependency with statistical difference （P〈0.05）. The levels of IL-4 and IL-5 in BALF of the 50 ng group, the 100 ng group and the 150 ng group were greatly decreased, showing statistical difference from those in the asthma group （P〈0.05） and negative dose dependency with statistical difference （P〈0.05）. The levels of IFN-γ of the 50 ng group, the 100 ng group and the 150 ng group were increased, showing statistical difference from those in the asthma group （P〈0.05） and negative dose dependency with statistical difference （P〈0.05）. The levels of serum IgE of the 50 ng group, the 100 ng group and the 150 ng group were greatly decreased, showing statistical difference from those in the asthma group （P〈0.05） and negative dose dependency with statistical difference （P〈0.05）. Conclusion VitD3 supply during maternal pregnancy alleviates offspring allergic airway inflammation of a mouse allergic asthma model in a dose-dependent manner.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2013年第22期2452-2455,共4页
Journal of Third Military Medical University
基金
杭州市科技计划项目(20100633B07)~~
