CAJ | 학술논문

目的观察静脉输注骨髓间充质干细胞(BMSC)对脊髓横断后大鼠下尿路功能的影响,并探讨其机制.方法获取SD大鼠BMSC,传代培养至5代时对BMSC行CD45和CD90检测,移植前将BMSC经3μg/ml的5-溴脱氧尿嘧啶(5-BrdU)共培养孵育3d,备移植用.采用随机数字表法将24只雄性SD大鼠分3组,每组8只.实验组大鼠建立脊髓横断模型后经尾静脉输注BMSC,对照组大鼠建立脊髓横断模型后经尾静脉输注培养基,假手术组大鼠仅作麻醉、切开切口及咬除椎板等操作.移植28 d后行尿动力学检查,取脊髓组织行HE染色病理检查及5-BrdU免疫组织化学检测.结果大鼠BMSC体外培养5代后,CD90阳性细胞达到99.4％,CD45阳性细胞数仅为5.94％.与对照组相比,实验组的残余尿量显著减少(1.54±0.66) ml和(0.85±0.16) ml,P＜0.01)],漏尿点压力显著降低(40.83±3.85) mmHg和(33.85±3.73) mmHg,P＜0.01)],逼尿肌无抑制性收缩次数显著减少9.71±3.84和2.17±1.12,P＜0.01)],排尿有效率显著增加(2.68±1.47)％和(26.37±6.97)％,P＜0.01)].对照组大鼠只见强直性肌电活动,脉冲性肌电活动消失,而实验组大鼠排尿期仍然可见这两种肌电图.与假手术组脉冲性肌电活动中静息电位时间(132.79±17.11) ms相比,实验组的(68.86±13.78) ms和脊髓损伤对照组(33.4) ms均显著变短(P＜0.01).免疫组织化学检测发现5-BrdU标记的BMSC除迁移于脊髓损伤区外,还定植于腰髓和骶髓.结论静脉输注的大鼠BSMC通过迁徙、定植过程促进脊髓损伤大鼠腰髓和骶髓形成新的排尿传导通路,减少导致膀胱功能障碍的神经突触形成,从而改善脊髓横断损伤大鼠的下尿路功能.
목적 관찰정맥수주골수간충질간세포(BMSC)대척수횡단후대서하뇨로공능적영향,병탐토기궤제.방법 획취SD대서BMSC,전대배양지5대시대BMSC행CD45화CD90검측,이식전장BMSC경3μg/ml적5-추탈양뇨밀정(5-BrdU)공배양부육3d,비이식용.채용수궤수자표법장24지웅성SD대서분3조,매조8지.실험조대서건립척수횡단모형후경미정맥수주BMSC,대조조대서건립척수횡단모형후경미정맥수주배양기,가수술조대서부작마취、절개절구급교제추판등조작.이식28 d후행뇨동역학검사,취척수조직행HE염색병리검사급5-BrdU면역조직화학검측.결과 대서BMSC체외배양5대후,CD90양성세포체도99.4％,CD45양성세포수부위5.94％.여대조조상비,실험조적잔여뇨량현저감소(1.54±0.66) ml화(0.85±0.16) ml,P＜0.01)],루뇨점압력현저강저(40.83±3.85) mmHg화(33.85±3.73) mmHg,P＜0.01)],핍뇨기무억제성수축차수현저감소9.71±3.84화2.17±1.12,P＜0.01)],배뇨유효솔현저증가(2.68±1.47)％화(26.37±6.97)％,P＜0.01)].대조조대서지견강직성기전활동,맥충성기전활동소실,이실험조대서배뇨기잉연가견저량충기전도.여가수술조맥충성기전활동중정식전위시간(132.79±17.11) ms상비,실험조적(68.86±13.78) ms화척수손상대조조(33.4) ms균현저변단(P＜0.01).면역조직화학검측발현5-BrdU표기적BMSC제천이우척수손상구외,환정식우요수화저수.결론 정맥수주적대서BSMC통과천사、정식과정촉진척수손상대서요수화저수형성신적배뇨전도통로,감소도치방광공능장애적신경돌촉형성,종이개선척수횡단손상대서적하뇨로공능.
Objective To determine whether intravenously transplanted bone marrow stromal cells (BMSCs) could promote recovery of lower urinary tract function in rats with spinal cord transaction.Method The BMSCs were harvested from rats and labeled with 5-bromo-2'-deoxyuridine (BrdU) (30 g/mL) in vitro for 3 days for intravenous transplantation.The BMSCs of 5th passage were characterized by flow cytometry of CD45 and CD90.Twenty-four male Sprague-Dawley rats were divided randomly into three goups.Sixteen rats received a spinal cord transection and a transplantation of cell culture medium (OP-controls,n =8) or BMSCs (n =8).Eight rats were justly given T8-T9 laminectomy (CNS-intact group,n =8).Urodynamic tests,HE staining and immunohistochemistry of the spinal cord with anti-BrdU at 28th day after transplantation were performed.Result The cultured BMSCs of 5th passage expressed 5.94％ of CD45 and 99.4％ of CD90 on flow cytometry.BMSC rats showed significant decrease in residual urine volume (0.85 ± 0.16 mL vs.1.54 ± 0.66 mL,P＜ 0.05),leak point pressure (33.85 ± 3.73 mmHg vs.40.83 ± 3.85 mmHg,P＜0.01),episodes of non-inhibitory contraction (NVC) (2.17 ± 1.12 vs.9.71 ± 3.84,P＜0.01) and increase in voiding efficiency (26.37 ± 6.97％ vs.2.68 ± 1.47,P＜0.01) compared to OP-control rats.In OP-control rats,only tonic EUS activity was remained and busting EUS activity was abolished during bladder filling.While,the tonic EUS activity and busting EUS activity were remained in BMSCs rats.Otherwise,the silent period between EUS bursts was shorter in BMSCs rats (68.86 ± 13.78 ms,P＜ 0.05) and OP-control rats (33.4 ms) than CNS-intact rats (132.79 ± 17.11 ms).Cells derived from BMSCs,identified by 5-BrdU immunoreactivity,survived and were distributed at the damaged tissue (T10),lumbar and sacral spinal cord.Conclusion Intravenously transplanted BMSCs survived in the injured spinal cord,lumbar and sacral cord for rebuilding new voiding reflex and inhibiting afferent sprouting leading to baldder dysfunction.So they had beneficial effects on the recovery of lower urinary tract function in rats with spinal cord transection.