中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2010年
1期
87-89,后插2
,共4页
杨新玲%马青平%贾丛康%张晨
楊新玲%馬青平%賈叢康%張晨
양신령%마청평%가총강%장신
帕金森病%胰岛素%骨髓基质细胞%移植
帕金森病%胰島素%骨髓基質細胞%移植
파금삼병%이도소%골수기질세포%이식
Parkinson's disease%Insulin%Bone marrow stromal cells%Transplantation
目的 观察胰岛素联合骨髓基质干细胞(BMSCs)治疗帕金森病(PD)模型鼠的疗效.方法 采用贴壁法培养、纯化BMSCs,流式细胞仪检测第3代BMSCs细胞的表面标志表达情况;将6-OHDA单侧损毁成功的PD模型大鼠21只随机分为3组,每组各7只,分别移植BMSCs 12 μl(细胞密度为1×10~5个/μl)、同数量的BMSCs+胰岛素50 nmol/L和生理盐水12μl于模型鼠右侧纹状体.在干预后4周分别观察模型大鼠阿朴吗啡诱导的旋转行为的变化,运用免疫荧光观察每组2只模型大鼠脑内BMSCs细胞存活情况以及高效液相HPLC法检测每组5只模型人鼠脑内双侧黑质-纹状体区DA含量的变化.结果 (1)第3代的BMSCs表面标志物CD29、CD34、CD44和CD45表达分别为97.1%、0.0%、99.0%和0.5%;(2)细胞移植治疗4周后联合组旋转次数较BMSCs组(P<0.05)和生理盐水组(P<0.01)明显减少;(3)免疫荧光观察模型鼠脑内BMSCs细胞存活情况:BrdU阳性细胞数量胰岛素+BMSCs组干预4周后BrdU阳性细胞数比BMSCs组明显增多(P<0.01);(4)HPLC结果示:胰岛素联合组模型鼠脑内黑质.纹状体区DA含量虽较其他两组增加,但差异无统计学意义(P>O.05).结论 胰岛素联合BMSCs移植治疗PD模型的疗效总体好于BMSCs组和生理盐水组.
目的 觀察胰島素聯閤骨髓基質榦細胞(BMSCs)治療帕金森病(PD)模型鼠的療效.方法 採用貼壁法培養、純化BMSCs,流式細胞儀檢測第3代BMSCs細胞的錶麵標誌錶達情況;將6-OHDA單側損燬成功的PD模型大鼠21隻隨機分為3組,每組各7隻,分彆移植BMSCs 12 μl(細胞密度為1×10~5箇/μl)、同數量的BMSCs+胰島素50 nmol/L和生理鹽水12μl于模型鼠右側紋狀體.在榦預後4週分彆觀察模型大鼠阿樸嗎啡誘導的鏇轉行為的變化,運用免疫熒光觀察每組2隻模型大鼠腦內BMSCs細胞存活情況以及高效液相HPLC法檢測每組5隻模型人鼠腦內雙側黑質-紋狀體區DA含量的變化.結果 (1)第3代的BMSCs錶麵標誌物CD29、CD34、CD44和CD45錶達分彆為97.1%、0.0%、99.0%和0.5%;(2)細胞移植治療4週後聯閤組鏇轉次數較BMSCs組(P<0.05)和生理鹽水組(P<0.01)明顯減少;(3)免疫熒光觀察模型鼠腦內BMSCs細胞存活情況:BrdU暘性細胞數量胰島素+BMSCs組榦預4週後BrdU暘性細胞數比BMSCs組明顯增多(P<0.01);(4)HPLC結果示:胰島素聯閤組模型鼠腦內黑質.紋狀體區DA含量雖較其他兩組增加,但差異無統計學意義(P>O.05).結論 胰島素聯閤BMSCs移植治療PD模型的療效總體好于BMSCs組和生理鹽水組.
목적 관찰이도소연합골수기질간세포(BMSCs)치료파금삼병(PD)모형서적료효.방법 채용첩벽법배양、순화BMSCs,류식세포의검측제3대BMSCs세포적표면표지표체정황;장6-OHDA단측손훼성공적PD모형대서21지수궤분위3조,매조각7지,분별이식BMSCs 12 μl(세포밀도위1×10~5개/μl)、동수량적BMSCs+이도소50 nmol/L화생리염수12μl우모형서우측문상체.재간예후4주분별관찰모형대서아박마배유도적선전행위적변화,운용면역형광관찰매조2지모형대서뇌내BMSCs세포존활정황이급고효액상HPLC법검측매조5지모형인서뇌내쌍측흑질-문상체구DA함량적변화.결과 (1)제3대적BMSCs표면표지물CD29、CD34、CD44화CD45표체분별위97.1%、0.0%、99.0%화0.5%;(2)세포이식치료4주후연합조선전차수교BMSCs조(P<0.05)화생리염수조(P<0.01)명현감소;(3)면역형광관찰모형서뇌내BMSCs세포존활정황:BrdU양성세포수량이도소+BMSCs조간예4주후BrdU양성세포수비BMSCs조명현증다(P<0.01);(4)HPLC결과시:이도소연합조모형서뇌내흑질.문상체구DA함량수교기타량조증가,단차이무통계학의의(P>O.05).결론 이도소연합BMSCs이식치료PD모형적료효총체호우BMSCs조화생리염수조.
Objective To observe the curative effectiveness of insulin in combination with bone marrow stromal cells (BMSCs) for parkinson' s disease (PD) rat models. Methods (1) From BMSCs cultivating pints of view, Adherent method was used to raise and repeatedly purify BMSCs. In addition flow cytometry was exploited to detect expression of CD29, CD34, CD44 and CD45, which all the markers exis ted on the cell' s surface of the BMSCs in the third generation; (2) The succeed PD rat models (n = 21) were randomizefy divided to three groups,which were employed as injected BMSCs (1.2×10~6 cells/each model) ,insulin (50 nmol/L) associates BMSCs and normal saline group (12 μl), respectively. Four weeks later,the rotating frequency was calculated and the cell' s survival was tested by the immunofluores cent method. The DA level in the brain (substantia nigra and striatum) was detected by HPLC. Results (1) CD29, CD34, CD44 and CD45 BMSCs in the third generation were expressed 97. 1% ,0. 0% , 99.0% ,0.5%. (2) The rotating frequencies in the insulin group combined BMSCs was much more de creased in comparson with the BMSCs (P <0. 05) and the normal saline group (P < 0. 01). (3) The cell' survival was to observe. BrdU positive cells in the group interfered by insulin combined BMSCs were much more than the BMSCs group (P <0. 01). (4) DA concentration in the group of insulin combined BMSCs were increased compared to the other 2 groups, however it did not have the statistical significance (P >0.05). Conclusion The curative effect of the insulin combined BMSCs interfered the rat model was better than BMSCs treated group alone. It is essencial to provide further evidence in vivo on insulin protect do paminergic neurons.
