中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2011年
6期
904-906
,共3页
唐礼功%谢森%陈琪%李志雄
唐禮功%謝森%陳琪%李誌雄
당례공%사삼%진기%리지웅
间充质干细胞%炎症%免疫抑制%FK506
間充質榦細胞%炎癥%免疫抑製%FK506
간충질간세포%염증%면역억제%FK506
Mesenchymal stem cells%Inflammation%Immunosuppression%FK506
目的 观察用免疫抑制剂FK506对干扰素(IFN)-γ和肿瘤坏死因子(TNF)-α诱导的间充质干细胞(MSCs)免疫抑制功能的影响及其作用机制.方法 将经过不同炎症因子预处理的小鼠MSCs与脾细胞共培养,并在FK506作用72 h后,以噻唑蓝(MTT)比色法检测脾细胞增殖;以流式细胞术检测脾细胞凋亡;以实时定量聚合酶链反应(Real-time PCR)法检测MSCs发挥免疫抑制作用的主要分子,观察FK506对其影响.结果 IFN-γ和TNF-α联合预处理后的MSCs可以显著抑制脾细胞增殖,促进脾细胞凋亡,凋亡率可达(53.5±2.5)%,明显高于IFN-γ或TNF-α预处理的MSCs组以及对照组(P＜0.05),并且IFN-γ和TNF-α仅可以显著上调MSCs中诱导型一氧化氮合酶(iNOS)的表达,MSCs的免疫抑制作用可以被iNOS特异性抑制剂1400W所拮抗;但FK506则可以抑制MSCs中iNOS的表达,使MSCs对脾细胞增殖的抑制作用显著降低,脾细胞凋亡减少,凋亡率降至(44.3±3.2)%(P＜0.05).结论 FK506通过抑制IFN-γ和TNF-α联合预处理的MSCs中iNOS表达从而抑制其免疫抑制功能.
目的 觀察用免疫抑製劑FK506對榦擾素(IFN)-γ和腫瘤壞死因子(TNF)-α誘導的間充質榦細胞(MSCs)免疫抑製功能的影響及其作用機製.方法 將經過不同炎癥因子預處理的小鼠MSCs與脾細胞共培養,併在FK506作用72 h後,以噻唑藍(MTT)比色法檢測脾細胞增殖;以流式細胞術檢測脾細胞凋亡;以實時定量聚閤酶鏈反應(Real-time PCR)法檢測MSCs髮揮免疫抑製作用的主要分子,觀察FK506對其影響.結果 IFN-γ和TNF-α聯閤預處理後的MSCs可以顯著抑製脾細胞增殖,促進脾細胞凋亡,凋亡率可達(53.5±2.5)%,明顯高于IFN-γ或TNF-α預處理的MSCs組以及對照組(P＜0.05),併且IFN-γ和TNF-α僅可以顯著上調MSCs中誘導型一氧化氮閤酶(iNOS)的錶達,MSCs的免疫抑製作用可以被iNOS特異性抑製劑1400W所拮抗;但FK506則可以抑製MSCs中iNOS的錶達,使MSCs對脾細胞增殖的抑製作用顯著降低,脾細胞凋亡減少,凋亡率降至(44.3±3.2)%(P＜0.05).結論 FK506通過抑製IFN-γ和TNF-α聯閤預處理的MSCs中iNOS錶達從而抑製其免疫抑製功能.
목적 관찰용면역억제제FK506대간우소(IFN)-γ화종류배사인자(TNF)-α유도적간충질간세포(MSCs)면역억제공능적영향급기작용궤제.방법 장경과불동염증인자예처리적소서MSCs여비세포공배양,병재FK506작용72 h후,이새서람(MTT)비색법검측비세포증식;이류식세포술검측비세포조망;이실시정량취합매련반응(Real-time PCR)법검측MSCs발휘면역억제작용적주요분자,관찰FK506대기영향.결과 IFN-γ화TNF-α연합예처리후적MSCs가이현저억제비세포증식,촉진비세포조망,조망솔가체(53.5±2.5)%,명현고우IFN-γ혹TNF-α예처리적MSCs조이급대조조(P＜0.05),병차IFN-γ화TNF-α부가이현저상조MSCs중유도형일양화담합매(iNOS)적표체,MSCs적면역억제작용가이피iNOS특이성억제제1400W소길항;단FK506칙가이억제MSCs중iNOS적표체,사MSCs대비세포증식적억제작용현저강저,비세포조망감소,조망솔강지(44.3±3.2)%(P＜0.05).결론 FK506통과억제IFN-γ화TNF-α연합예처리적MSCs중iNOS표체종이억제기면역억제공능.
Objective To investigate the mechanism of immunosuppressant FK506 on the immunesuppression of mesenchymal stem cells ( MSCs) induced by interferon ( IFN) -γ and tumor necrosis factor (TNF)-α. Methods MSCs pretreated with inflammatory cytokines were co-cultured with mice spleen cells. After incubation with FK506 for 72 h, proliferation of spleen cells was measured by methyl thiazol tetrazolium (MTT) assay and the apoptosis was assessed by flow cytometry. Real-time polymerase chain reaction (PCR) was used to detect the major molecule that mediated the immunosuppression of MSCs and the effect of FK506 on the molecule. Results MSCs pretreated with inflammatory cytokines could inhibit the proliferation of spleen cells and increase the apoptosis of spleen cells, and the apoptosis rate was (53. 5 ±2. 5) % , which was higher than that of MSCs pretreated with IFN-γ or TNF-α and control group ( P ＜0. 05), and IFN-γ and TNF-α upregulated inducible nitric oxide synthase (iNOS) expression in MSCs significantly, and this immunosuppression could also be antagonized by a specific inhibitor 1400W. However,FK506 could inhibit iNOS expression in MSCs pretreated with IFN-γ and TNF-α, the inhibition of spleen cells proliferation by MSCs was reduced significantly, and the apoptosis rate of spleen cells was reduced to (44. 3 ±3. 2)% (P＜0. 05). Conclusion FK506 inhibits the immunosuppression of MSCs through downregulation of iNOS expression in MSCs pretreated with IFN-γ and TNF-α.
