中华风湿病学杂志
中華風濕病學雜誌
중화풍습병학잡지
CHINESE JOURNAL OF RHEUMATOLOGY
2011年
11期
763-766
,共4页
张继云%刘蕾%孟德芳%汤郁%施冬艳%王丹丹%刘慧青%李霞%孙凌云
張繼雲%劉蕾%孟德芳%湯鬱%施鼕豔%王丹丹%劉慧青%李霞%孫凌雲
장계운%류뢰%맹덕방%탕욱%시동염%왕단단%류혜청%리하%손릉운
红斑狼疮,系统性%细胞运动%细胞增殖%间质干细胞%IκB激酶
紅斑狼瘡,繫統性%細胞運動%細胞增殖%間質榦細胞%IκB激酶
홍반랑창,계통성%세포운동%세포증식%간질간세포%IκB격매
Lupus erythematosus,systemic%Cell movement%Cell proliferation%Mesenchymalstemcells%IKK-β
目的 观察系统性红斑狼疮(SLE)患者骨髓间充质干细胞(BMSCs)迁移、增殖变化,并探讨IKB激酶复合物(IKK-β)对SLE患者BMSCs迁移及增殖的影响.方法 采用密度梯度离心和贴壁分离法分离培养6例SLE患者及6名健康人BMSCs；通过细胞划痕实验和构建侵袭小室检测BMSCs迁移、活细胞计数(CCK-8)检测BMSCs增殖能力；实时荧光定量聚合酶链反应技术检测IKK-β mRNA表达水平；蛋白印迹法检测IKK-β、磷酸化IKK-β表达水平；观察加入IKK-β抑制剂对SLE患者BMSCs增殖、迁移的影响.采用t检验或Mann-Whitney秩和检验.结果 ①体外培养过程中,SLE患者BMSCs迁移率(5.2±3.8)‰与增殖能力(0.21±0.49)显著低于健康对照组BMSCs迁移率(7.0±2.9)‰及增殖能力(1.00±0.35),差异有统计学意义(P＜0.05);②SLE患者BMSCs IKK-β基因表达水平(1.9±1.4)与健康人(1.9±2.4)相比差异无统计学意义(P＞0.05)；SLE患者BMSCs磷酸化IKK-β蛋白表达水平(1.41±0.19)显著高于健康人(0.93±1.24),差异有统计学意义(P＜0.05)；③IKK-β抑制剂能够增加SLE患者BMSCs的迁移率(3.3±1.6)‰和增殖能力(1.13±0.26),高于未加IKK-β抑制剂组迁移率(2.3±1.1)‰与增殖能力(0.8l±0.17),差异有统计学意义(P＜0.05).结论 SLE患者BMSCs在体外培养过程中,迁移、增殖能力较健康人显著降低,其机制可能是SLE患者体内IKK-β的表达增加,通过活化核因子-KB信号通路,影响迁移与增殖相关基因的表达,从而抑制了BMSCs的迁移与增殖.
目的 觀察繫統性紅斑狼瘡(SLE)患者骨髓間充質榦細胞(BMSCs)遷移、增殖變化,併探討IKB激酶複閤物(IKK-β)對SLE患者BMSCs遷移及增殖的影響.方法 採用密度梯度離心和貼壁分離法分離培養6例SLE患者及6名健康人BMSCs；通過細胞劃痕實驗和構建侵襲小室檢測BMSCs遷移、活細胞計數(CCK-8)檢測BMSCs增殖能力；實時熒光定量聚閤酶鏈反應技術檢測IKK-β mRNA錶達水平；蛋白印跡法檢測IKK-β、燐痠化IKK-β錶達水平；觀察加入IKK-β抑製劑對SLE患者BMSCs增殖、遷移的影響.採用t檢驗或Mann-Whitney秩和檢驗.結果 ①體外培養過程中,SLE患者BMSCs遷移率(5.2±3.8)‰與增殖能力(0.21±0.49)顯著低于健康對照組BMSCs遷移率(7.0±2.9)‰及增殖能力(1.00±0.35),差異有統計學意義(P＜0.05);②SLE患者BMSCs IKK-β基因錶達水平(1.9±1.4)與健康人(1.9±2.4)相比差異無統計學意義(P＞0.05)；SLE患者BMSCs燐痠化IKK-β蛋白錶達水平(1.41±0.19)顯著高于健康人(0.93±1.24),差異有統計學意義(P＜0.05)；③IKK-β抑製劑能夠增加SLE患者BMSCs的遷移率(3.3±1.6)‰和增殖能力(1.13±0.26),高于未加IKK-β抑製劑組遷移率(2.3±1.1)‰與增殖能力(0.8l±0.17),差異有統計學意義(P＜0.05).結論 SLE患者BMSCs在體外培養過程中,遷移、增殖能力較健康人顯著降低,其機製可能是SLE患者體內IKK-β的錶達增加,通過活化覈因子-KB信號通路,影響遷移與增殖相關基因的錶達,從而抑製瞭BMSCs的遷移與增殖.
목적 관찰계통성홍반랑창(SLE)환자골수간충질간세포(BMSCs)천이、증식변화,병탐토IKB격매복합물(IKK-β)대SLE환자BMSCs천이급증식적영향.방법 채용밀도제도리심화첩벽분리법분리배양6례SLE환자급6명건강인BMSCs；통과세포화흔실험화구건침습소실검측BMSCs천이、활세포계수(CCK-8)검측BMSCs증식능력；실시형광정량취합매련반응기술검측IKK-β mRNA표체수평；단백인적법검측IKK-β、린산화IKK-β표체수평；관찰가입IKK-β억제제대SLE환자BMSCs증식、천이적영향.채용t검험혹Mann-Whitney질화검험.결과 ①체외배양과정중,SLE환자BMSCs천이솔(5.2±3.8)‰여증식능력(0.21±0.49)현저저우건강대조조BMSCs천이솔(7.0±2.9)‰급증식능력(1.00±0.35),차이유통계학의의(P＜0.05);②SLE환자BMSCs IKK-β기인표체수평(1.9±1.4)여건강인(1.9±2.4)상비차이무통계학의의(P＞0.05)；SLE환자BMSCs린산화IKK-β단백표체수평(1.41±0.19)현저고우건강인(0.93±1.24),차이유통계학의의(P＜0.05)；③IKK-β억제제능구증가SLE환자BMSCs적천이솔(3.3±1.6)‰화증식능력(1.13±0.26),고우미가IKK-β억제제조천이솔(2.3±1.1)‰여증식능력(0.8l±0.17),차이유통계학의의(P＜0.05).결론 SLE환자BMSCs재체외배양과정중,천이、증식능력교건강인현저강저,기궤제가능시SLE환자체내IKK-β적표체증가,통과활화핵인자-KB신호통로,영향천이여증식상관기인적표체,종이억제료BMSCs적천이여증식.
Objective To investigate the effect of IκB kinase (IKK-β) on migration and proliferation of bone marrow mesenchymal stem cells (BMSCs) from patients with systemic lupus erythematosus (SLE).Methods Human bone marrow aspirates were collected from iliac of six donors and six SLE patients and cultured in vitro.Migration of BMSCs were observed by wound healing and transwell migration assays.Proliferation of BMSCs was quantified by cell counting kit-8 assay.Total RNA was extracted and reverse transcribed into complementary DNA.Real-time PCR technique was used to determine the gene expression of IKK-β at transcription level.The expression of IKK-β and phospho-IKK-β (p-IKK-β) protein were determined by Western blotting analysis.Statistical analysis was conducted with or Mann-Whitney rank test.Results ① The migration rate of BMSCs from SLE patients (5.2±3.8)‰ were significantly reduced as compared with normal controls (7.0±2.9)‰(P＜0.05 ).The proliferation of BMSCs of SLE patients (0.21±0.49)was lower than that from healthy controls ( 1.00±0.35 )(P＜0.05 ).②) No difference in IKK-β3 mRNA expression between SLE ( 1.9± 1.4) subjects and normal controls (1.9±2.4) (P＞0.05).IKK-β protein expression of BMSCs from SLE patients (1.41 ±0.19) increased significantly compared with healthy controls (0.93±1.24) (P＜0.05).③ Inhibitor of IKK-β caused a significant increase in cell migration (3.3±1.6)‰ and proliferation (1.13±0.26) of BMSCs from SLE patients compared with untreated cells (2.3±1.1)‰ and (0.81±0.17),respectively (P＜0.05).Conclusion Migration and proliferation of BMSCs are significantly decreased in SLE patients.IKK-β may be involved in migration and proliferation of BMSCs.
