中华内分泌代谢杂志
中華內分泌代謝雜誌
중화내분비대사잡지
CHINESE JOURNAL OF ENDOCRINOLOGY AND METABOLISM
2012年
6期
467-471
,共5页
罗建华%李占伟%黄海%杨冬花%于瑞萍%喻日成%范元硕%刘波
囉建華%李佔偉%黃海%楊鼕花%于瑞萍%喻日成%範元碩%劉波
라건화%리점위%황해%양동화%우서평%유일성%범원석%류파
糖尿病,实验性%小鼠,近交NOD%细胞凋亡%PPARγ%NF-κB
糖尿病,實驗性%小鼠,近交NOD%細胞凋亡%PPARγ%NF-κB
당뇨병,실험성%소서,근교NOD%세포조망%PPARγ%NF-κB
Diabetes mellitus,experimeatal%Mice,inbred NOD%Apoptosis%PPARγ%NF-κB
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-κB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/ml,P=0.045];在培养的脾细胞上清中,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PPARγ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PPARγ,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β细胞凋亡减少.
目的 探討吡格列酮預防非肥胖糖尿病小鼠胰島β細胞凋亡的機製.方法 (1)將4週齡NOD雌鼠分為吡格列酮(21隻)及對照(21隻)組,分彆攝食含0.02％吡格列酮的混閤飼料和普通營養飼料.觀察52週齡的纍積糖尿病髮病率.(2)各組取12週齡未髮病NOD鼠(n=15)胰腺,HE染色觀察胰島炎;TUNEL+SABC法檢測胰島β細胞凋亡.(3)ELISA法測定血清、脾細胞培養上清IFN-γ和IL-4水平及培養脾細胞覈因子PPARγ、NF-κB活性.結果 (1)30、52週齡時,吡格列酮及對照組髮病率分彆為57.1％和76.2％ 、76.2％和90.5％(均P＞0.05)；15週齡時,吡格列酮及對照組髮病率分彆為4.8％和33.3％(P=0.045).(2) 12週齡時,吡格列酮組正常胰島和胰島週圍炎比例(14.73％,26.02％)高于對照組(5.69％,15.72％；均P＜0.01),胰島內炎比例(59.25％)則低于對照組(78.59％,P＜0.01)；吡格列酮組胰島β細胞凋亡率(6.17％±3.62％)低于對照組( 10.62％±4.43％,P=0.008).(3)12週齡NOD鼠吡格列酮組血清IFN-γ水平[(561.05 ±78.61) pg/ml]顯著低于對照組[(666.43±28.42) pg/ml,P=0.045];在培養的脾細胞上清中,吡格列酮組IFN-γ水平[(605.84±65.60) pg/ml]顯著低于對照組[(692.20±44.98)pg/ml,P=0.041].(4)在培養的脾細胞中,吡格列酮組PPARγ活性(0.06±0.01)高于對照組(0.03±0.01,P=0.013),NF-κB活性(0.03±0.01)較對照顯著降低(0.08±0.01,P=0.001).結論 吡格列酮活化PPARγ,抑製NF-κB活性,血清和脾細胞上清IFN-γ下降,Th細胞嚮Th1方嚮分化減少,NOD鼠胰島炎減輕、胰島β細胞凋亡減少.
목적 탐토필격렬동예방비비반당뇨병소서이도β세포조망적궤제.방법 (1)장4주령NOD자서분위필격렬동(21지)급대조(21지)조,분별섭식함0.02％필격렬동적혼합사료화보통영양사료.관찰52주령적루적당뇨병발병솔.(2)각조취12주령미발병NOD서(n=15)이선,HE염색관찰이도염;TUNEL+SABC법검측이도β세포조망.(3)ELISA법측정혈청、비세포배양상청IFN-γ화IL-4수평급배양비세포핵인자PPARγ、NF-κB활성.결과 (1)30、52주령시,필격렬동급대조조발병솔분별위57.1％화76.2％ 、76.2％화90.5％(균P＞0.05)；15주령시,필격렬동급대조조발병솔분별위4.8％화33.3％(P=0.045).(2) 12주령시,필격렬동조정상이도화이도주위염비례(14.73％,26.02％)고우대조조(5.69％,15.72％；균P＜0.01),이도내염비례(59.25％)칙저우대조조(78.59％,P＜0.01)；필격렬동조이도β세포조망솔(6.17％±3.62％)저우대조조( 10.62％±4.43％,P=0.008).(3)12주령NOD서필격렬동조혈청IFN-γ수평[(561.05 ±78.61) pg/ml]현저저우대조조[(666.43±28.42) pg/ml,P=0.045];재배양적비세포상청중,필격렬동조IFN-γ수평[(605.84±65.60) pg/ml]현저저우대조조[(692.20±44.98)pg/ml,P=0.041].(4)재배양적비세포중,필격렬동조PPARγ활성(0.06±0.01)고우대조조(0.03±0.01,P=0.013),NF-κB활성(0.03±0.01)교대조현저강저(0.08±0.01,P=0.001).결론 필격렬동활화PPARγ,억제NF-κB활성,혈청화비세포상청IFN-γ하강,Th세포향Th1방향분화감소,NOD서이도염감경、이도β세포조망감소.
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 splenocyte,the activity of PPARγ and NF-κB 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 infiltrated 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 (692.20+44.98) pg/ml,P=0.041].(4) In cultured splenocyte,PPARγ nuclear protein activity in pioglitazone group was higher than that in control group ( 0.06 ± 0.01 vs 0.03 ± 0.01,P =0.013 ),and NF-κB nuclear protein activity was lower than that in control group ( 0.03 ± 0.01 vs 0.08± 0.01,P =0.001 ).Conclusions Pioglitazone activates PPARγ nuclear protein,inhibits activity of NF-κB nuclear protein,downregulates IFN-γ,diminishes differeutiation of Th cells to Th1,and subsequently prevents insulitis and β-cell apoptosis in NOD mice.
