中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2011年
2期
209-211
,共3页
李煜环%宋振顺%范子扬%张福琴
李煜環%宋振順%範子颺%張福琴
리욱배%송진순%범자양%장복금
糖尿病%骨髓间充质干细胞%T细胞%免疫调节
糖尿病%骨髓間充質榦細胞%T細胞%免疫調節
당뇨병%골수간충질간세포%T세포%면역조절
Diabetes mellitus%Bone marrow mesenchymal stem cells%T cells%Immunoregulation
目的 观察骨髓间充质干细胞(MSCs)体外对1型糖尿病(T1DM)大鼠淋巴细胞表型及增殖能力的影响,探讨其抑制淋巴细胞增殖的机制.方法 分离、培养和鉴定大鼠MSCs,噻唑蓝(MIT)比色法观察该细胞对淋巴细胞增殖能力的影响,应用流式细胞术分析MSCs对植物血凝素(PHA)作用下淋巴细胞凋亡,周期水平和CD4+CD25+调节性T细胞亚群(CD4+CD25+Tregs)比例的影响.结果 大鼠MSCs表型为CD29+、CD90+、CD106+、CD34-、CD45-,对PHA刺激的淋巴细胞增殖有抑制作用,以淋巴细胞:MSCs为1∶1时(C组)抑制作用最强;共培养体系中,大部分淋巴细胞处于G0/G1期;C组淋巴细胞凋亡水平(58.05±0.89)%显著高于对照组(43.35±0.86)%(P＜0.05);CD4+CD25+Tregs的比例C组(22.76±1.15)%显著高于对照组(5.80±0.68)%(P＜0.05).结论 MSCs体外可显著抑制PHA刺激的T1DM大鼠淋巴细胞的增殖,其机制与CD4+CD25+Tregs比例增高密切相关.
目的 觀察骨髓間充質榦細胞(MSCs)體外對1型糖尿病(T1DM)大鼠淋巴細胞錶型及增殖能力的影響,探討其抑製淋巴細胞增殖的機製.方法 分離、培養和鑒定大鼠MSCs,噻唑藍(MIT)比色法觀察該細胞對淋巴細胞增殖能力的影響,應用流式細胞術分析MSCs對植物血凝素(PHA)作用下淋巴細胞凋亡,週期水平和CD4+CD25+調節性T細胞亞群(CD4+CD25+Tregs)比例的影響.結果 大鼠MSCs錶型為CD29+、CD90+、CD106+、CD34-、CD45-,對PHA刺激的淋巴細胞增殖有抑製作用,以淋巴細胞:MSCs為1∶1時(C組)抑製作用最彊;共培養體繫中,大部分淋巴細胞處于G0/G1期;C組淋巴細胞凋亡水平(58.05±0.89)%顯著高于對照組(43.35±0.86)%(P＜0.05);CD4+CD25+Tregs的比例C組(22.76±1.15)%顯著高于對照組(5.80±0.68)%(P＜0.05).結論 MSCs體外可顯著抑製PHA刺激的T1DM大鼠淋巴細胞的增殖,其機製與CD4+CD25+Tregs比例增高密切相關.
목적 관찰골수간충질간세포(MSCs)체외대1형당뇨병(T1DM)대서림파세포표형급증식능력적영향,탐토기억제림파세포증식적궤제.방법 분리、배양화감정대서MSCs,새서람(MIT)비색법관찰해세포대림파세포증식능력적영향,응용류식세포술분석MSCs대식물혈응소(PHA)작용하림파세포조망,주기수평화CD4+CD25+조절성T세포아군(CD4+CD25+Tregs)비례적영향.결과 대서MSCs표형위CD29+、CD90+、CD106+、CD34-、CD45-,대PHA자격적림파세포증식유억제작용,이림파세포:MSCs위1∶1시(C조)억제작용최강;공배양체계중,대부분림파세포처우G0/G1기;C조림파세포조망수평(58.05±0.89)%현저고우대조조(43.35±0.86)%(P＜0.05);CD4+CD25+Tregs적비례C조(22.76±1.15)%현저고우대조조(5.80±0.68)%(P＜0.05).결론 MSCs체외가현저억제PHA자격적T1DM대서림파세포적증식,기궤제여CD4+CD25+Tregs비례증고밀절상관.
Objective To observe the effects of bone marrow mesenchymal stem cells (MSCs) on the lymphocytes of rats with type 1 diabetes mellitus (T1DM) in vitro, and investigate the inhibitory effect of MSCs on lymphocytes proliferation and the underlying mechanism. Methods MSCs were isolated from SD rats, cultured in vitro, purified and then identified by testing the phenotypes with flow cytometry (FCM). The third-generation MSCs were planted in 24-well plates. After treated with mitomycin C, MSCs were co-cultured for 72 h with the T1 DM rat's lymphocytes activated by phytohemagglutinin (PHA). The proliferation of lymphocyte was measured by methyl thiazol tetrazolium (MTT) method. FCM analysis was done to investigate the apoptosis, cell cycle and the proportion of CD4+ CD25+ regulatory T cells of the T1 DM rat's lymphocytes after co-cultivation. Results The phenotypes of MSCs from normal SD rats were CD29 + , CD90 +, CD106 + , CD34-, CD45 -. MSCs obviously inhibited the lymphocyte proliferation stimco-culture system, most of the lymphocytes were arrested at G0/G1 phase. The apoptosis rate of lymphocytes (58.05 ± 0. 89)% in group C was increased significantly as compared with the control group (43.35± 0.86 ) % ( P ＜ 0. 05 ) as well as the proportion of CD4 + CD25 + regulatory T cells (22.76 ± 1.15 ) % vs (5.80 ± 0. 68) %. Conclusion In vitro, MSCs can obviously inhibit the T1 DM rat' s lymphocytes proliferation stimulated with PHA via increasing the proportion of CD4 + CD25 + regulatory T cells.
