中华核医学杂志
中華覈醫學雜誌
중화핵의학잡지
CHINESE JOURNAL OF NUCLEAR MEDICINE
2010年
3期
180-184
,共5页
胡硕%兰晓莉%曹卫%曹国祥%张国鹏%张斌青%吴涛%常伟%张永学
鬍碩%蘭曉莉%曹衛%曹國祥%張國鵬%張斌青%吳濤%常偉%張永學
호석%란효리%조위%조국상%장국붕%장빈청%오도%상위%장영학
大鼠%心肌%骨髓%干细胞%移植%钠碘转运体%放射性核素显像
大鼠%心肌%骨髓%榦細胞%移植%鈉碘轉運體%放射性覈素顯像
대서%심기%골수%간세포%이식%납전전운체%방사성핵소현상
Rats%Myocardium%Bone marrow%Stem cells%Transplantation%Sodium/iodide symporter%Radionuclide imaging
目的 研究大鼠钠碘转运体(rNIS)作为报告基因监测大鼠骨髓间充质干细胞(rBMSC)移植至大鼠心肌的可行性.方法 构建含rNIS/增强绿色荧光蛋白(EGFP)基因的重组腺病毒载体(Ad/rNIS/EGFP),转染rBMSC后应用荧光显微镜观察细胞表达EGFP情况.按随机数字表法将健康大鼠分为3组,rNIS组大鼠心肌内移植转染rNIS的rBMSC;抑制组大鼠心肌内移植转染rNIS的rBMSC,γ显像前口服高氯酸纳;对照组大鼠心肌内移植未转染病毒的rBMSC.细胞移植到大鼠心肌后,以99TcmO4-作为报告探针对大鼠进行γ显像,随后进行生物学分布实验,用实时聚合酶链反应(PCR)和Western blot分别检测心肌组织中rNIS基因和蛋白表达水平,免疫组织化学检测心肌组织中CD11b、CD29、CD34、CD44、CD45和CD90表达情况.组间计量资料的差异用t检验分析.结果 重组腺病毒转染后,荧光显微镜下可见到高表达EGFP的rBMSC.rNIS转染的rBMSC移植到大鼠心肌后,γ显像能清晰显示移植部位的心肌,心肌/右前肢放射性比值(RUR)为6.7±0.4,对照组RUR为3.0±0.2,二者之间比较差异有统计学意义,t=2.78,P=0.03.生物学分布实验中,对照组心肌组织的每克组织百分注射剂量率(%ID/g)为2.5±0.4,明显低于rNIS组的60.2±20.8,差异有统计学意义,t=7.13,P＜0.001.移植心肌组织中rNIS的基因和蛋白都高表达,而对照组心肌组织rNIS表达较低.移植转染细胞的心肌组织中,CD29、CD44、CD90表达阳性,CD11b表达弱阳性,CD34、CD45表达阴性.结论 rNIS可作为报告基因有效监测rBMSC移植大鼠心肌.
目的 研究大鼠鈉碘轉運體(rNIS)作為報告基因鑑測大鼠骨髓間充質榦細胞(rBMSC)移植至大鼠心肌的可行性.方法 構建含rNIS/增彊綠色熒光蛋白(EGFP)基因的重組腺病毒載體(Ad/rNIS/EGFP),轉染rBMSC後應用熒光顯微鏡觀察細胞錶達EGFP情況.按隨機數字錶法將健康大鼠分為3組,rNIS組大鼠心肌內移植轉染rNIS的rBMSC;抑製組大鼠心肌內移植轉染rNIS的rBMSC,γ顯像前口服高氯痠納;對照組大鼠心肌內移植未轉染病毒的rBMSC.細胞移植到大鼠心肌後,以99TcmO4-作為報告探針對大鼠進行γ顯像,隨後進行生物學分佈實驗,用實時聚閤酶鏈反應(PCR)和Western blot分彆檢測心肌組織中rNIS基因和蛋白錶達水平,免疫組織化學檢測心肌組織中CD11b、CD29、CD34、CD44、CD45和CD90錶達情況.組間計量資料的差異用t檢驗分析.結果 重組腺病毒轉染後,熒光顯微鏡下可見到高錶達EGFP的rBMSC.rNIS轉染的rBMSC移植到大鼠心肌後,γ顯像能清晰顯示移植部位的心肌,心肌/右前肢放射性比值(RUR)為6.7±0.4,對照組RUR為3.0±0.2,二者之間比較差異有統計學意義,t=2.78,P=0.03.生物學分佈實驗中,對照組心肌組織的每剋組織百分註射劑量率(%ID/g)為2.5±0.4,明顯低于rNIS組的60.2±20.8,差異有統計學意義,t=7.13,P＜0.001.移植心肌組織中rNIS的基因和蛋白都高錶達,而對照組心肌組織rNIS錶達較低.移植轉染細胞的心肌組織中,CD29、CD44、CD90錶達暘性,CD11b錶達弱暘性,CD34、CD45錶達陰性.結論 rNIS可作為報告基因有效鑑測rBMSC移植大鼠心肌.
목적 연구대서납전전운체(rNIS)작위보고기인감측대서골수간충질간세포(rBMSC)이식지대서심기적가행성.방법 구건함rNIS/증강록색형광단백(EGFP)기인적중조선병독재체(Ad/rNIS/EGFP),전염rBMSC후응용형광현미경관찰세포표체EGFP정황.안수궤수자표법장건강대서분위3조,rNIS조대서심기내이식전염rNIS적rBMSC;억제조대서심기내이식전염rNIS적rBMSC,γ현상전구복고록산납;대조조대서심기내이식미전염병독적rBMSC.세포이식도대서심기후,이99TcmO4-작위보고탐침대대서진행γ현상,수후진행생물학분포실험,용실시취합매련반응(PCR)화Western blot분별검측심기조직중rNIS기인화단백표체수평,면역조직화학검측심기조직중CD11b、CD29、CD34、CD44、CD45화CD90표체정황.조간계량자료적차이용t검험분석.결과 중조선병독전염후,형광현미경하가견도고표체EGFP적rBMSC.rNIS전염적rBMSC이식도대서심기후,γ현상능청석현시이식부위적심기,심기/우전지방사성비치(RUR)위6.7±0.4,대조조RUR위3.0±0.2,이자지간비교차이유통계학의의,t=2.78,P=0.03.생물학분포실험중,대조조심기조직적매극조직백분주사제량솔(%ID/g)위2.5±0.4,명현저우rNIS조적60.2±20.8,차이유통계학의의,t=7.13,P＜0.001.이식심기조직중rNIS적기인화단백도고표체,이대조조심기조직rNIS표체교저.이식전염세포적심기조직중,CD29、CD44、CD90표체양성,CD11b표체약양성,CD34、CD45표체음성.결론 rNIS가작위보고기인유효감측rBMSC이식대서심기.
Objective To investigate the feasibility of rat sodium/iodide symporter (rNIS) as a reporter gene monitoring rat bone marrow mesenchymal cells (rBMSC) transplanted to rat myocardium in vivo.Methods Recombinated adenovirus vector was constructed by rNIS/enhanced green fluorescence protein (EGFP) (Ad-rNIS/EGFP).rBMSC transfected by Ad-rNIS/EGFP were studied using fluorescence microscope.Fifteen rats were transplanted with rBMSC and randomly divided into three groups:rNIS group (with rNIS transfection), blocked group (with rNIS transfection) by oral intake of perchloric sodium before planar imaging(GE Millennium MPR SPECT), and control group (without rNIS transfection).All rats underwent 99Tcm-pertechnetate planar imaging.The biological distribution of 99Tcm-pertechnetate was studied.The expressions of rNIS gene and protein in myocardium were measured by real time polymerase chain reaction (PCR) and western blot, respectively.The expressions of CD29, CD44, CD90, CD11b, CD34 and CD45 were measured by immunohistochemistry.Results rBMSC transfected by Ad-rNIS/EGFP showed EGFP expression under fluorescence microscope.The transplanted rat myocardium could be visualized on 99Tcm-pertechnetate planar imaging in rNIS group.The relative uptake ratio( Rheart/Rhmb, RUR) was 6.7 ±0.4.RUR in control group (3.0 ±0.2) was lower than that in rNIS group (t =2.78, P=0.03).The percentage injection dose per gram of tissue (% ID/g) of the transplanted myocardium was 60.2 ± 20.8 in rNIS group,which was higher than that (2.5 ± 0.4) % ID/g of control group ( t = 7.13, P＜0.001 ).rNIS gene and protein were highly expressed in transplanted myocardium in rNIS group but less expressed in control group.The expressions of CD29, CD44 and CD90 were positive, CD45 and CD45 negative CD11b mildly positive in the myocardium transplanted with infective rBMSC.Conclusion rNIS can efficiently monitor rBMSC transplanted to rat myocardium.
