维生素D3补充对大鼠免疫细胞功能影响的实验研究

目的：维生素D（VD）对机体免疫功能作用已引起广泛关注，本实验研究补充不同剂量VD，对大鼠免疫细胞活性的影响。方法：将30只Wistar大鼠随机分为3组，分别为VD，1（对照组）、VD，2和VD，3组，每组10只，雌雄各半。VD，l组大鼠每天摄入VD，20IU／kg（生理剂量，相当于成人每天摄入5txg维生素D，）、VD2组和VD3组分别补充200IU／（kg·d）（10倍量）和1000IU／（kg·d）（50倍量）的VD，；补充时间为4周。实验结束时麻醉无痛处死，取血样（约5mL）离心，采用LC-Ms／MS测定血清VD，浓度、CCK-8法检测淋巴细胞增殖活性、流式细胞术测单核细胞吞噬功能。结果：经过4周VD，补充后，结果显示，VD，1、VD，2和VD。3组大鼠血清VD，浓度分别为14．8ng／mL、34．7n【g／mL和60．7ng／mL，随着补充剂量的增加，机体VD，吸收水平也显著提高；VDl补充后，VD，2组淋巴细胞增殖活性的OD值达到3．04，明显高于VD，1组的2．64（P〈0．05），但VD33组与VD31组相比淋巴细胞增殖活性无显著性差异（P〉0．05）。采用流式细胞术观察单核细胞吞噬活性（荧光强度），结果显示VD，2组大鼠单核细胞吞噬活性荧光强度到达19．98，明显高于VD，1组的15．48，而高剂量VD，补充的VD，3组大鼠单核细胞吞噬活性未见明显增强。结论：适宜剂量VD，200IU／（kg·d）]补充可增强淋巴细胞增殖和单核细胞吞噬活性，而过高剂量（50倍生理剂量）未观察到明显免疫调节作用。

Effect of Different Doses of Vitamin D3 Supplementation on Activities of Lymphocytes and Mononuclear Cells in Rats

Objective] The effect of vitamin D （VD） on immune function has attracted wide attention. The objective of this experi- ment is to study the effect of different doses of vitamin D3 （ VD3 ） supplementation on immune cells activities in rats. Method] Totally 30 Wistar rats were randomly divided into 3 groups： group VD3 1 （control group）, group VD3 2 and group VD3 3. Each group had 10 rats, male and female in equal. The daily intake of VD3 in group VD3 1 was 20 IU/ （ kg . d） （ equal to 5 μg/d vitamin D3 3 for adults）, the do- ses of VD3 supplemented to group VD3 2 and group VD3 3 were 200IU/ （ kg.d） and 1 000IU/ （ kg.d） respectively. The supplementation period was 4 weeks. At the end of the trial, rats were anesthetized to painless death, samples of whole blood （about 5mL） were collected and centrifuged. LC-MS/MS was used to detect serum vitamin D3. CCK - 8 method was used to detect lymphocyte proliferation activity and flow cytometry （FCM） was used to detect phagocytosis function of mononuclear cell. Result] After 4 weeks supplementation of VD3, ser- um vitamin D concentration in VD3 1, VD3 2 and VD3 3 groups were 14. 8, 34. 7, and 60. 7 ng/mL respectively. Along with the increasing of supplementation doses, the amount of absorbed VD3 also increased remarkably. After supplementation of VD3, lymphocyte proliferation activity （OD） in group VD3 2 was 3.04 significantly higher than that of group VD3 1 which was 2.64 （ P 〈 0. 05 ）, while there were no sig- nificant differences between group VD3 3 and group VD3 1 （P 〉 0. 05 ） . Phagocytosis function of mononuclear cell （fluorescence intensity） were detected by FCM. The results showed that fluorescence intensity of group VD3 2 was 19. 98 significantly higher than that of group VD3 1 which was 15.48, while there were no significant increase of phagocytosis function of mononuclear cell in group VD3 3. Conclusion] Appro- priate dose of VD3 200IU/ （kg.d） ] supplementation might enhance lymphocyte proliferation activity and phagocytic function of mononu- clear cells, while exorbitant doses supplementation （50-fold physiological dose of VD3 ） failed to observe significant improvement in im- mune function.