中华风湿病学杂志
中華風濕病學雜誌
중화풍습병학잡지
CHINESE JOURNAL OF RHEUMATOLOGY
2011年
7期
468-471,后插1
,共5页
刘蕾%孟德芳%汤郁%张继云%施冬艳%王丹丹%孙凌云
劉蕾%孟德芳%湯鬱%張繼雲%施鼕豔%王丹丹%孫凌雲
류뢰%맹덕방%탕욱%장계운%시동염%왕단단%손릉운
红斑狼疮,系统性%细胞凋亡%间质干细胞
紅斑狼瘡,繫統性%細胞凋亡%間質榦細胞
홍반랑창,계통성%세포조망%간질간세포
Lupus erythematosus,systemic%Apoptosis%Mesenchymal stem cells
目的 探讨系统性红斑狼疮(SLE)患者骨髓间充质干细胞(BMSCs)的凋亡变化及凋亡相关因子的表达.方法 密度梯度离心和贴壁筛选法分离培养SLE患者及健康人骨髓MSCs,末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(TUNEL)法检测细胞凋亡;流式细胞术检测细胞表面Fas、Bcl-2表达水平及Caspase 8活性;实时荧光定量聚合酶链反应(PCR)技术榆测Fas、Bcl-2、Bax、Bcl-w、Caspase 8及凋亡细胞蛋白酶激活因子(Apaf)-1 mRNA表达水平;免疫组织化学染色检测胞质细胞色素C表达水平;统计学采用Mann-Whitnev秩和检验.结果 SLE患者BMSCs凋亡明显高于健康对照组[(64±10)%和(14±9)%,U=0,P<0.05];SLE患者BMSCs抗凋亡因子Bcl-2蛋白表达显著低于健康对照组[(11±9)%和(56±18)%,U=0,P<0.05],其mRNA表达水平同样显著低于健康对照(0.2±0.2和2.4±0.7,U=24.P<0.05);SLE患者BMSCs胞质中细胞色素C表达显著增多[(56±21)%和(16±16)%,U=1,P<0.05].SLE患者BMSCs细胞内Caspase 8活性增强[(49±14)%和(16±12)%,U=0,P<0.05],但基因表达水平与健康对照组比较差异无统计学意义(U=28,P>0.05);SLE患者与健康对照组BMSCs均高表达Fas,差异无统计学意义(U=19,P>0.05).结论 SLE患者BMSCs凋亡增多,可能与Bcl-2表达减少、胞质中细胞色素C表达增多和Caspase 8活性增强,从而激活内源性及外源性凋亡途径相关.
目的 探討繫統性紅斑狼瘡(SLE)患者骨髓間充質榦細胞(BMSCs)的凋亡變化及凋亡相關因子的錶達.方法 密度梯度離心和貼壁篩選法分離培養SLE患者及健康人骨髓MSCs,末耑脫氧覈苷痠轉移酶介導的dUTP缺口末耑標記(TUNEL)法檢測細胞凋亡;流式細胞術檢測細胞錶麵Fas、Bcl-2錶達水平及Caspase 8活性;實時熒光定量聚閤酶鏈反應(PCR)技術榆測Fas、Bcl-2、Bax、Bcl-w、Caspase 8及凋亡細胞蛋白酶激活因子(Apaf)-1 mRNA錶達水平;免疫組織化學染色檢測胞質細胞色素C錶達水平;統計學採用Mann-Whitnev秩和檢驗.結果 SLE患者BMSCs凋亡明顯高于健康對照組[(64±10)%和(14±9)%,U=0,P<0.05];SLE患者BMSCs抗凋亡因子Bcl-2蛋白錶達顯著低于健康對照組[(11±9)%和(56±18)%,U=0,P<0.05],其mRNA錶達水平同樣顯著低于健康對照(0.2±0.2和2.4±0.7,U=24.P<0.05);SLE患者BMSCs胞質中細胞色素C錶達顯著增多[(56±21)%和(16±16)%,U=1,P<0.05].SLE患者BMSCs細胞內Caspase 8活性增彊[(49±14)%和(16±12)%,U=0,P<0.05],但基因錶達水平與健康對照組比較差異無統計學意義(U=28,P>0.05);SLE患者與健康對照組BMSCs均高錶達Fas,差異無統計學意義(U=19,P>0.05).結論 SLE患者BMSCs凋亡增多,可能與Bcl-2錶達減少、胞質中細胞色素C錶達增多和Caspase 8活性增彊,從而激活內源性及外源性凋亡途徑相關.
목적 탐토계통성홍반랑창(SLE)환자골수간충질간세포(BMSCs)적조망변화급조망상관인자적표체.방법 밀도제도리심화첩벽사선법분리배양SLE환자급건강인골수MSCs,말단탈양핵감산전이매개도적dUTP결구말단표기(TUNEL)법검측세포조망;류식세포술검측세포표면Fas、Bcl-2표체수평급Caspase 8활성;실시형광정량취합매련반응(PCR)기술유측Fas、Bcl-2、Bax、Bcl-w、Caspase 8급조망세포단백매격활인자(Apaf)-1 mRNA표체수평;면역조직화학염색검측포질세포색소C표체수평;통계학채용Mann-Whitnev질화검험.결과 SLE환자BMSCs조망명현고우건강대조조[(64±10)%화(14±9)%,U=0,P<0.05];SLE환자BMSCs항조망인자Bcl-2단백표체현저저우건강대조조[(11±9)%화(56±18)%,U=0,P<0.05],기mRNA표체수평동양현저저우건강대조(0.2±0.2화2.4±0.7,U=24.P<0.05);SLE환자BMSCs포질중세포색소C표체현저증다[(56±21)%화(16±16)%,U=1,P<0.05].SLE환자BMSCs세포내Caspase 8활성증강[(49±14)%화(16±12)%,U=0,P<0.05],단기인표체수평여건강대조조비교차이무통계학의의(U=28,P>0.05);SLE환자여건강대조조BMSCs균고표체Fas,차이무통계학의의(U=19,P>0.05).결론 SLE환자BMSCs조망증다,가능여Bcl-2표체감소、포질중세포색소C표체증다화Caspase 8활성증강,종이격활내원성급외원성조망도경상관.
Objective To investigate the apoptosis of bone marrow mesenchymal stem cells(BMSCs)from systemic lupus erythematosus(SLE)patients and the expression of apoptotic molecules.Methods BMSCs were isolated from bone marrow of SLE patients and normal controls by density eentrifugation and adhesive culture in vitro.The apoptosis of BMSCs were evaluated by TUNEL assay.The expressions of Fas.Bcl-2 and the activity of Caspase 8 were detected by flow cytometry.Real-time PCR technique was used to determine the gene expressions of Fas,Bcl-2,Bax,Bcl-w,Caspase 8 and Apaf-1.Meanwhile,cytochrome C was detected by immunocytochemistry.Statistical analysis was conducted with t-test and Mann-Whitney rank test.Results The percentage of apoptotic BMSCs increased in SLE patients compared with healthy donors[(64±10)%vs [14±9)%,U=0,P<0.05 ].The expression of Bcl-2 in BMSCs of SLE patients was lower than the normal controls at protein leve[(11±9)%vs(56±18)%,U=0,P<0.05 ],and mRNA level(0.2±0.2vs 2.4±0.7,U=24,P<0.05).More cytochrome C positive pellets in the cytosolic fraction could be detected in BMSCs from SLE patients compared with healthy controls [(56±21)%vs (16±16)%,U=1,P<0.05].The activity of Caspase 8 was enhanced[(49±14)%vs(16±12)%,U=0.P<0.05 ],although with no significant difierence at mRNA Ievel.Both groups expressed Fas but with no significant difference (U=19,P>0.05).Conclusion BMSCs from SLE patients undergo more apoptosis,the mechanisms may be associated with the down regulation of Bcl-2,up-regulation of Cytoehrome C in cytoplasm and the activation of Caspase 8,which directs the intrinsic and extrinsic apoptosis pathways.
