目的 研究给予母孕期大鼠补充不同剂量维生素D,检测子代鼠胸腺中VDR表达变化。方法 健康雌性Wistar大鼠18只,雄性Wistar大鼠9只,以每日晚间17时将雌鼠、雄鼠按2∶1合笼,次日8时对雌鼠阴道分泌物涂片,在40倍光学显微镜检查到精子表示受孕,记为妊娠0 d( gestational day 0,GD0) 。将孕鼠随机入对照组、维生素D低剂量、维生素D高剂量组。于GD 15 d起对照组每日给予肌肉注射生理盐水0.25 ml/(kg·次),低剂量组给予肌肉注射维生素D 20 μg/kg,高剂量组给予肌肉注射维生素D 100 μg/kg,直至子鼠娩出。子鼠哺乳21 d后分离子鼠胸腺,Real-time PCR检测VDRmRNA表达水平,Western blot检测VDR蛋白表达水平。结果 通过Real-tine PCR分析仔鼠胸腺VDR基因表达三组之间差异无统计学意义(P＞0.05)。Western blot检测VDR蛋白水平的表达显示,低剂量及高剂量组VDR蛋白的表达量均高于对照组,三组之间差异存在统计学意义(P＜0.05)。结论 给予母鼠补充维生素D,其仔鼠胸腺VDR蛋白表达量明显增加,与维生素D补充量呈正相关。
Abstract
Objective To detect the changes of vitamin D receptor (VDR) expression in the thymus of filial generation mice after giving pregnant mice different dosages of vitamin D supplementation. Methods Totally 18 female Wistar mice mates with 9 male Wistar mice at 5 pm everyday (the female/male radio was 2∶1).The vaginal secretions of the female mice were collected and made into smears.Conception was recorded after finding sperm in the smears,noting gestational day 0 (GD0).The pregnant mice were separated into three groups:control group,low level of vitamin D supplementation group and high level of vitamin D supplementation group.Different dosages of vitamin D supplementation were given by intramuscular injection from GD 15,with normal saline 0.25 ml/kg,vitamin D 20 μg/kg and vitamin D 100 μg/kg separately for each group.The injection continued until delivery.After 21 days breast-feeding,the thymus of filial generation mice were isolated.The expression of VDRmRNA was detected by Real-time PCR and the expression of VDR protein was detected by Western blot. Results No statistic difference of VDR gene expression was observed among different groups by Real-time PCR(P＞0.05).Western blot showed that the expression of VDR protein in low level of vitamin D supplementation group and high level of vitamin D supplementation group was higher than that of the control group (P＜0.05). Conclusion The expression of VDR in filial generation mice increases if mother mice are given vitamin D supplementation during pregnancy and is positively related to the dosage of vitamin D supplementation.
关键词
维生素D /
维生素D受体 /
胸腺
Key words
vitamin D /
vitamin D receptor /
thymus
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参考文献
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基金
黑龙江省自然科学基金项目 (H201397)
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