中华风湿病学杂志
中華風濕病學雜誌
중화풍습병학잡지
CHINESE JOURNAL OF RHEUMATOLOGY
2014年
3期
155-159,后插1
,共6页
高晋芳%张莉芸%李小峰%辛磊%杨艳丽
高晉芳%張莉蕓%李小峰%辛磊%楊豔麗
고진방%장리예%리소봉%신뢰%양염려
关节炎,类风湿%间质干细胞%增殖能力%端粒酶逆转录酶
關節炎,類風濕%間質榦細胞%增殖能力%耑粒酶逆轉錄酶
관절염,류풍습%간질간세포%증식능력%단립매역전록매
Arthritis,rheumatoid%Bone mesenchymal stem cells%Proliferation capability%Human telomerase reverse transcriptase
目的 比较RA患者与健康人骨髓间充质干细胞(BMSCs)的增殖能力及端粒酶反转录酶(hTERT)的表达,探讨RA患者BMSCs是否存在缺陷或变异,能否用于自体移植.方法 取8例RA患者与8名健康志愿者5 ml骨髓,分离、培养、传代,用流式细胞术对其表面标记进行鉴定,四甲基偶氮唑蓝(MTT)法测定P3、P6代BMSCs生长曲线,计算细胞群体倍增时间;流式细胞术测定细胞周期,计算增殖指数;实时荧光定量PCR测定hTERT的表达.组间数据比较采用t检验.结果 RA患者BMSCs在形态学上与健康对照组无差异.经流式细胞仪检测,证明获得细胞为BMSCs.健康对照组与RA组P3代细胞群体倍增时间(4.7±0.3)d,(4.8±0.3)d,2组差异无统计学意义(P＞0.05);P6代健康对照组细胞群体倍增时间(5.7±0.4)d,RA组(6.3±0.3)d,RA组BMSCs群体倍增时间长于健康对照组,差异有统计学意义(P＜0.001).健康对照组P3代BMSCs增殖指数(17.9±2.1)％,RA组(16.9±2.1)％,2组差异无统计学意义(P＞0.05);P6代健康对照组BMSCs增殖指数(10.7±1.8)％,RA组(6.2±2.2)％,RA组增殖指数小于对照组,差异有统计学意义(P＜0.01).健康对照组P6代MSCs hTERT表达量为(10.6±1.5),RA组为(3.1±1.0),RA患者hTERT表达水平较正常对照组低,差异有统计学意义(P＜0.01).结论 2组人群P3代BMSCs细胞增殖能力的差异无统计学意义,P6代时RA患者BMSCs增殖能力滞后于正常组,推断RA患者BMSCs较健康人衰老速度加快;hTERT测定显示RA组BMSCs的hTERT表达量低于正常对照组,说明这种提前老化可能与hTERT表达过低有关.
目的 比較RA患者與健康人骨髓間充質榦細胞(BMSCs)的增殖能力及耑粒酶反轉錄酶(hTERT)的錶達,探討RA患者BMSCs是否存在缺陷或變異,能否用于自體移植.方法 取8例RA患者與8名健康誌願者5 ml骨髓,分離、培養、傳代,用流式細胞術對其錶麵標記進行鑒定,四甲基偶氮唑藍(MTT)法測定P3、P6代BMSCs生長麯線,計算細胞群體倍增時間;流式細胞術測定細胞週期,計算增殖指數;實時熒光定量PCR測定hTERT的錶達.組間數據比較採用t檢驗.結果 RA患者BMSCs在形態學上與健康對照組無差異.經流式細胞儀檢測,證明穫得細胞為BMSCs.健康對照組與RA組P3代細胞群體倍增時間(4.7±0.3)d,(4.8±0.3)d,2組差異無統計學意義(P＞0.05);P6代健康對照組細胞群體倍增時間(5.7±0.4)d,RA組(6.3±0.3)d,RA組BMSCs群體倍增時間長于健康對照組,差異有統計學意義(P＜0.001).健康對照組P3代BMSCs增殖指數(17.9±2.1)％,RA組(16.9±2.1)％,2組差異無統計學意義(P＞0.05);P6代健康對照組BMSCs增殖指數(10.7±1.8)％,RA組(6.2±2.2)％,RA組增殖指數小于對照組,差異有統計學意義(P＜0.01).健康對照組P6代MSCs hTERT錶達量為(10.6±1.5),RA組為(3.1±1.0),RA患者hTERT錶達水平較正常對照組低,差異有統計學意義(P＜0.01).結論 2組人群P3代BMSCs細胞增殖能力的差異無統計學意義,P6代時RA患者BMSCs增殖能力滯後于正常組,推斷RA患者BMSCs較健康人衰老速度加快;hTERT測定顯示RA組BMSCs的hTERT錶達量低于正常對照組,說明這種提前老化可能與hTERT錶達過低有關.
목적 비교RA환자여건강인골수간충질간세포(BMSCs)적증식능력급단립매반전록매(hTERT)적표체,탐토RA환자BMSCs시부존재결함혹변이,능부용우자체이식.방법 취8례RA환자여8명건강지원자5 ml골수,분리、배양、전대,용류식세포술대기표면표기진행감정,사갑기우담서람(MTT)법측정P3、P6대BMSCs생장곡선,계산세포군체배증시간;류식세포술측정세포주기,계산증식지수;실시형광정량PCR측정hTERT적표체.조간수거비교채용t검험.결과 RA환자BMSCs재형태학상여건강대조조무차이.경류식세포의검측,증명획득세포위BMSCs.건강대조조여RA조P3대세포군체배증시간(4.7±0.3)d,(4.8±0.3)d,2조차이무통계학의의(P＞0.05);P6대건강대조조세포군체배증시간(5.7±0.4)d,RA조(6.3±0.3)d,RA조BMSCs군체배증시간장우건강대조조,차이유통계학의의(P＜0.001).건강대조조P3대BMSCs증식지수(17.9±2.1)％,RA조(16.9±2.1)％,2조차이무통계학의의(P＞0.05);P6대건강대조조BMSCs증식지수(10.7±1.8)％,RA조(6.2±2.2)％,RA조증식지수소우대조조,차이유통계학의의(P＜0.01).건강대조조P6대MSCs hTERT표체량위(10.6±1.5),RA조위(3.1±1.0),RA환자hTERT표체수평교정상대조조저,차이유통계학의의(P＜0.01).결론 2조인군P3대BMSCs세포증식능력적차이무통계학의의,P6대시RA환자BMSCs증식능력체후우정상조,추단RA환자BMSCs교건강인쇠로속도가쾌;hTERT측정현시RA조BMSCs적hTERT표체량저우정상대조조,설명저충제전노화가능여hTERT표체과저유관.
Objective By comparing the proliferation and the human telomerase reverse transcriptase expression of bone marrow mesenchymal stem cells (BMSCs) from rheumatoid arthritis (RA) patients and normal human to explore the existence of BMSCs defects or variation in RA patients,and whether these cells could be used in autologous transplantation.Methods BMSCs were obtained from bone marrow of eight RA patients and healthy volunteers.The cells were isolated andcultured by gradient centrifugation and adherence cultivation,and the surface markers of BMSCs were identitfied by flow cytometertry.Firstly,BMSCs reproductive activity of the two groups were compared:the auxodrome and cell cycle of BMSCs were measured respectively,then the generation doubling time of cell colony and proliferation index were calculated in order to see whether there was a difference of the proliferation capability between the two groups.Furthermore,the hTERT were measured.Results BMSCs were no significant differences between morphology of RA patients and healthy control group.The cells were obtained proof MSCs by flow cytometry.The difference was not statistically significant (P＞0.05),RA group and the healthy control group P3-generation cell population doubling time (4.7±0.3) d,(4.8±0.3) d.The difference was statistically significant (P＜0.01),P6 generation of healthy control group cell population has doubling time (5.7±0.4) d,RA group (6.3±0.3) d,buut the group RA BMSCs population doubling time is longer than the healthy control group.The difference was not statistically significant (P＞0.05),on one hand of the healthy control group P3 BMSCs proliferation index (17.9±2.1)％,on the other hand RA group is (16.9±2.1)％.The difference was statistically significant (P＜0.01).P6-generation BMSCs proliferation index in healthy control group (10.7±1.8)％,RA group (6.2±2.2)％,RA group proliferation index than the control group.The difference was also significant (P＜0.01).The healthy control group P6 generation of MSCs hTERT expression was (10.6±1.5),RA group is (3.1±1.0),hTERT RA patients expression is lower compared with normal control group.Conclusion There is no significant difference in the proliferative capacity of the two groups at P3,but the BMSCs proliferation at P6 lags behind in the normal group in RA patients,which inferred that the senescences of the BMSCs of RA patients are earlier than healthy donors.The determination results of hTERT have shown that the difference between the two groups is statistically significant,and it proves that the premature aging may be associated with hTERT expression.
