CAJ | 학술논문

目的：观察非肥胖性糖尿病（ non-obese diabetic， NOD）小鼠自然病程不同阶段CD19＋CD5＋CD1dhigh B细胞及Th1、Th17细胞数量的变化，探讨B10细胞与NOD小鼠自身免疫性糖尿病发生的关系。方法采用流式细胞术分别检测4周（A组，n＝10）、8周（B组，n＝10）及糖尿病（C组，n＝10）NOD小鼠及同龄C57BL／6小鼠（对照组，n＝20）脾脏、肠系膜淋巴结、外周淋巴结、胰腺淋巴结中CD19＋CD5＋CD1dhigh B、CD19＋IL-10＋B、CD4＋IFN-γ＋Th1、CD4＋IL-17＋Th17细胞及CD4＋CD25＋Foxp3＋T细胞的比例；分离NOD小鼠胰腺组织作病理切片以评估胰岛炎的严重程度。结果（1）胰腺组织HE切片：A组无胰岛炎，4周；B组发生胰岛炎，8周；C组发生明显胰岛炎且无完整胰岛，糖尿病。（2）不同病程阶段NOD小鼠B10细胞比较：B及C组B10细胞较A组增高，而C组B10细胞则较B组显著下降，差异有统计学意义（P＜0．01）；A组B10细胞各组织间无明显差异（P＞0．05），B组B10细胞以胰腺淋巴结最高，C组B10细胞以外周淋巴结最高且胰腺淋巴结B10细胞较B组显著下降，差异有统计学意义（P＜0．01）。（3）不同病程阶段NOD小鼠效应性T细胞比较：C组Th1及Th17细胞较A、B组显著增加，差异有统计学意义（P＜0．01）。（4）不同病程阶段CD4＋CD25＋Foxp3＋T细胞比较：C组Treg较A、B组无明显变化，且NOD小鼠Treg较同龄C57BL／6小鼠亦无明显变化（ P＞0．05）。结论随NOD小鼠年龄增长及胰岛炎进展，B10细胞存在数量变化及组织分布的差异；B10细胞数量减少可能参与NOD小鼠自身免疫性糖尿病的发生。
목적：관찰비비반성당뇨병（ non-obese diabetic， NOD）소서자연병정불동계단CD19＋CD5＋CD1dhigh B세포급Th1、Th17세포수량적변화，탐토B10세포여NOD소서자신면역성당뇨병발생적관계。방법채용류식세포술분별검측4주（A조，n＝10）、8주（B조，n＝10）급당뇨병（C조，n＝10）NOD소서급동령C57BL／6소서（대조조，n＝20）비장、장계막림파결、외주림파결、이선림파결중CD19＋CD5＋CD1dhigh B、CD19＋IL-10＋B、CD4＋IFN-γ＋Th1、CD4＋IL-17＋Th17세포급CD4＋CD25＋Foxp3＋T세포적비례；분리NOD소서이선조직작병리절편이평고이도염적엄중정도。결과（1）이선조직HE절편：A조무이도염，4주；B조발생이도염，8주；C조발생명현이도염차무완정이도，당뇨병。（2）불동병정계단NOD소서B10세포비교：B급C조B10세포교A조증고，이C조B10세포칙교B조현저하강，차이유통계학의의（P＜0．01）；A조B10세포각조직간무명현차이（P＞0．05），B조B10세포이이선림파결최고，C조B10세포이외주림파결최고차이선림파결B10세포교B조현저하강，차이유통계학의의（P＜0．01）。（3）불동병정계단NOD소서효응성T세포비교：C조Th1급Th17세포교A、B조현저증가，차이유통계학의의（P＜0．01）。（4）불동병정계단CD4＋CD25＋Foxp3＋T세포비교：C조Treg교A、B조무명현변화，차NOD소서Treg교동령C57BL／6소서역무명현변화（ P＞0．05）。결론수NOD소서년령증장급이도염진전，B10세포존재수량변화급조직분포적차이；B10세포수량감소가능삼여NOD소서자신면역성당뇨병적발생。
Objective To study the alterations of CD19+CD5+CD1dhigh B, Th1 and Th17 cells in non-obese diabetic ( NOD) mice and the correlation between B10 cells and type 1 diabetes in NOD mice. Methods Flow cytometry ( FCM) was used to measure the levels of CD19+CD5+CD1dhigh B, CD19+IL-10+B, CD4+IFN-γ+Th1, CD4+IL-17+Th17 and CD4+CD25+Foxp3+T cells in NOD mice ( 4 weeks old NOD mice:group A, n=10;8 weeks old NOD mice:group B, n=10; NOD mice with diabetes: group C, n=10) and age-matched C57BL/6 mice ( control group, n=20 ) .Hematoxylin-eosin staining of pancreatic tissues was performed for histopathological assessment of the development of insulitis in NOD mice.Results (1) Histopathological analysis showed that mice from A, B and C groups respectively showed no insulitis, insulitis and obvious insulitis with no intact islets.(2) The highest levels of B10 cells in NOD mice were ob-served in group B, followed by those in group C and group A (P<0.01).No significant differences with the levels of B10 cells were found among different tissues of 4 weeks old NOD mice (P>0.05).More B10 cells were detected in pancreatic lymph nodes than in other tissues of 8 weeks old NOD mice, the levels of which were also higher than those in pancreatic lymph nodes of mice form group C ( P<0.01) .The highest levels of B10 cells were detected in peripheral lymph nodes among all tissues samples collected from NOD mice with diabetes (P<0.01).(3) The levels of Th1 and Th17 cells in mice from group C were remarkably in-creased as compared with those in mice from group A and B (P<0.01).(4) The percentages of CD4+CD25+Foxp3+T cells in mice from group C showed no differences with those in mice from A and B groups. No significant difference with Treg cells were observed between NOD mice and age-matched C57BL/6 mice (P>0.05).Conclusion The percentages and distribution of B10 cells in NOD mice changed with age and the development of insulitis.The decrease of B10 cells might participate in the development of type 1 diabe-tes in NOD mice.