磷酸二酯酶抑制剂Rolipram对NOD小鼠免疫应答的影响  

The role of Rolipram in immune response in NOD mice

作　　者：路文盛[1] 师布朵 王鲁梅[3] 张志利[4] 

机构地区：[1]中山大学附属第二医院内分泌科,广州510120 [2]山西省同德医院内分泌科 [3]广东省东莞市人民医院皮肤科 [4]山西医科大学第一临床医学院内分泌研究所

出　　处：《中华微生物学和免疫学杂志》2005年第5期405-410,共6页Chinese Journal of Microbiology and Immunology

摘　　要：目的　探讨磷酸二酯酶Ⅳ型抑制剂Rolipram对非肥胖糖尿病(NOD)小鼠免疫应答的影响机制,为临床通过免疫干预治疗糖尿病提供理论依据。方法　动物模型分为Rolipram干预组和磷酸盐缓冲液(PBS)对照组,分别经DNAladder、免疫组化检测T细胞凋亡;流式细胞、RT- PCR测定T细胞亚群的漂移。结果　与PBS对照组比较,Rolipram干预组外周T淋巴细胞凋亡率增加(Bcl- 2表达减少:0 .3439±0 .0 739vs 0 .736 1±0 .0 839,t=2 .6 82 ,P <0 .0 1;Bax表达增加:0 .7380±0 .1372vs 0 .32 2 7±0 .0 4 2 8,t=2 .6 79,P <0 .0 1;同时DNAladder也呈典型的2 0 0bp及其倍数大小的条带) ;CD4 + TH1型淋巴细胞减少(IL- 2A2 6 0 βactinA2 6 0 :0 .0 33±0 .0 0 1vs 0 .2 87±0 .0 0 6 ,t =2 .70 1,P <0 .0 1)而CD4 + TH2型细胞增加(IL 10A2 6 0 βactinA2 6 0 :0 .32 3±0 .0 0 4vs 0 .0 72±0 .0 0 3,t=2 .135 ,P <0 .0 5 ) ;胰岛炎症积分明显减少(Z =- 1.882 ,P <0 .0 1)。结论　磷酸二酯酶Ⅳ型抑制剂Rolipram能引起T细胞功能抑制,减轻NOD小鼠自身免疫反应程度,保护胰岛β细胞功能,其机制可能是通过T细胞亚群由TH1型向TH2型定向漂移实现的。Objective To study the effect of Rolipram in n on -obese diabetic mice by means of DNA ladder, immunohistochemical method, flow c ytometry quantitive analysis and reverse transcript PCR technology. M ethods Ninety female NOD/Lt mice(20 g) were randomly divided into two groups. Treatment group received intraperitoneal injection of Rolipram (8 mg /kg) twice a day. The control group received intraperitoneal injection of PBS. A ll experimental mice got intraperitoneal injection of cyclophosphamide (200 mg/k g) to accelerate the process of diabetes on day 1 and 15. All mice were killed o n day 30 to study different expression of Bcl-2/Bax in lymph and pancreas, and the distribution of T lymphocytes in thymus and spleen, the apoptosis rate of T lymphocytes in spleen, which was detected by flow cytometry quantitive analysis and DNA ladder qualitative analysis technology. The subpopulation shift of T H 1/T H2 was detected by reverse transcript PCR technology. Results The ratio of Bcl-2/Bax was higher in pancreasthan in lymph (P<0. 01); the apoptosis rate of T lymphocytes and the quantity of CD3+(P<0.01), CD4+ T cells and IL-10 lymphocyte in spleen was higher(P<0.05); the quan tity of CD8+ T cells was invariable; IL-2 lymphocyte in spleen was decreased (P<0.01); the number of CD4+CD8- T cells and CD4-CD8+ T cells increa sed markedly (P<0.01), and CD4-CD8- double negative T cells and CD4+CD 8+ double positive T cells were invariable in thymus. Conclusion Rolipram can prevent NOD/Lt mice from developing into type 1 diabet es by increasing apoptosis of T lymphocyte or by increased shifting from CD4+ T H1 to CD4+ T H2 cells; decrease expression of Bcl-2 in lymphatic tissue, on the contrary, increase it in pancreatic tissue; increase CD4+CD8- and d ecrease CD4-CD8+ in thymus and increase CD3+, CD4+ in spleen. However, C D4+CD8+, CD4-CD8- and CD8+ aren′t associated with Rolipram. Rolipram can increase expression of IL-10 mRNA and decrease IL-2 mRNA to protect pancre atic islets β cells.

关 键 词：磷酸二酯酶抑制剂 ROLIPRAM NOD小鼠 免疫应答 糖尿病 免疫干预 治疗 

分 类 号：R96[医药卫生—药理学]