吡格列酮预防非肥胖糖尿病鼠胰岛β细胞凋亡的机制研究  

Prevention of islet β-cell apoptosis by pioglitazone in NOD mice and its mechanism

作　　者：罗建华[1] 李占伟[3] 黄海[3] 杨冬花 于瑞萍[1] 喻日成[1] 范元硕[1] 刘波[1] 

机构地区：[1]贵州省人民医院内分泌科,贵阳550002 [2]贵州省人民医院干医科,贵阳550002 [3]贵阳医学院附属医院检验科

出　　处：《中华内分泌代谢杂志》2012年第6期467-471,共5页Chinese Journal of Endocrinology and Metabolism

基　　金：贵州省优秀科技教育人才省长专项基金[黔省专合字（2007）60号];贵州省社会发展攻关项目（黔科合SY[2008]3051号）;贵州省高层次人才科研特助基金（TZJF-2007年-47号）

摘　　要：目的探讨吡格列酮预防非肥胖糖尿病小鼠胰岛β细胞凋亡的机制。方法（1）将4周龄NOD雌鼠分为吡格列酮（21只）及对照（21只）组，分别摄食含0．02％吡格列酮的混合饲料和普通营养饲料。观察52周龄的累积糖尿病发病率。（2）各组取12周龄未发病NOD鼠（n=15）胰腺，HE染色观察胰岛炎；TUNEL＋SABc法检测胰岛β细胞凋亡。（3）ELISA法测定血清、脾细胞培养上清IFN-γ和IL-4水平及培养脾细胞核因子PPARγ、NF—kB活性。结果（1）30、52周龄时，吡格列酮及对照组发病率分别为57．1％和76．2％、76．2％和90．5％（均P〉0．05）；15周龄时，吡格列酮及对照组发病率分别为4．8％和33．3％（P=0．045）。（2）12周龄时，吡格列酮组正常胰岛和胰岛周围炎比例（14．73％，26．02％）高于对照组（5．69％，15．72％；均P〈0．01），胰岛内炎比例（59．25％）则低于对照组（78．59％，P〈0．01）；吡格列酮组胰岛β细胞凋亡率（6．17％±3．62％）低于对照组（10．62％±4．43％，P=0．008）。（3）12周龄NOD鼠吡格列酮组血清IFN-γ水平[（561．05±78．61）pg/ml]显著低于对照组[（666．43±28．42）pg／m1，P=0．045]；在培养的脾细胞上清巾，吡格列酮组IFN-γ水平[（605．84±65．60）pg/ml]显著低于对照组[（692．20±44．98）pg／ml，P=0．041]。（4）在培养的脾细胞中，吡格列酮组PPA脚活性（0．06±0．01）高于对照组（0．03±0．01，P=0．013），NF—κB活性（0．03±0．01）较对照显著降低（0．08±0．01，P=0．001）。结论吡格列酮活化PPAR吖，抑制NF—κB活性，血清和脾细胞上清IFN-γ下降，Th细胞向Th1方向分化减少，NOD鼠胰岛炎减轻、胰岛8细胞捌亡减少。Objective To investigate the mechanism of preventing islet β-cell apoptosis in NOD mice with pioglitazone. Methods Female NOD mice at 4 weeks of age were divided into pioglitazone group （ n = 21, 0.02% pioglitazone was added into the feed） and control group （ n = 21, fed with regular diet）. The accumulative incidence of diabetes was followed-up to 52 weeks of age in each group of NOD mice. Pancreas was removed from NOD mice at 12 weeks of age in each group （ n = 15 ） to score severity of insulitis by routine H-E staining. The apoptotic β-cells in islets were observed with double-labeling technique of TUNEL in situ combined with standard sensitive avidin-biotin complex （sABC） immunohistochemical method. The spleens were taken for cell culture; IL-4 and IFN-γ levels in sera and supernatants of cultured splenoeyte, the activity of PPAR＇y and NF-KB nuclear proteins in cultured splenocyte were measured by ELISA. Results （ 1 ） At 30 and 52 weeks of age, the respective incidences of diabetes were 57. 1% and 76.2% in pioglitazone group, and 76.2% and 90.5% in control group （ all P〉0. 05 ）. At 15 weeks of age, the incidence became 4.8% in pioglitazone group, and 33.3% in control group （ P = 0. 045 ）. （2） At 12 weeks of age, the percentages of non infihrated islet and peri-insulitis islet in pioglitazone group were higher than those in control group （ 14.73% vs 5.69% , P〈0. 01; and 26.02% vs 15.72% , P〈0. 01 ） , and that of intraislet insulitis was lower than that in control group （59.25% vs 78.59% , P〈0. 01 ）. The percentage of apoptotic β-cell in pioglitazone group was lower＇ than that in control group （ 6.17 % ± 3.62 % vs 10.62 % ±4.43 %, P = 0. 008 ）. （ 3 ） In sera, IFN-γ, level in pioglitazone group was lower than that in control group [ （ 561.05 ± 78.61 ） vs （ 666.43 ± 28.42 ） pg/ml, P = 0. 045 ]. In cultured splenocyte supernatant, the level of IFN-γ in pioglitazone group was lower than that in control group[ （605.84± 65.60 ） vs （69

关 键 词：糖尿病 实验性 小鼠 近交NOD 细胞凋亡 PPARΓ NF—κB 

分 类 号：R587.1[医药卫生—内分泌]