CAJ | 학술논문

目的观察人雪旺细胞(hSCs)对人脐带间充质干细胞(hUC-MSCs)在脊髓损伤(SCI)体内存活、分化的影响,通过分子生物学技术检测胶质纤维酸性蛋白(GFAP)、髓磷脂碱性蛋白(MBP)与高分子量神经丝蛋白(NF-H)的表达,描述其变化的时间特征.方法分离、培养及纯化hUC-MSCs和hSCs.取8周龄雌性Wistar大鼠60只,以Impactor Model-Ⅱ型打击器制作脊髓胸10(T10)损伤模型.大鼠随机均分为4组:DMEM对照组(A)、hSCs移植组(B)、hUC-MSCs移植组(C)、hUC-MSCs与hSCs联合移植组(D),各组分别于移植后1、2、3、4周取材.结果 D组可见MBP与核因子(NF)-H染色阳性细胞,其他各组为阴性.A组3种神经标志物在蛋白与mRNA水平的表达量随时间延长均逐渐增高,且GFAP在移植后2周明显增高,MBP与NF-H在移植后3周明显增高[2周与1周GFAP条带灰度比值为1.34,而聚合酶链反应(PCR)所测比值为2.86；3周与1周MBP和NF-H条带灰度比值分别为3.43、4.12,而PCR所测比值分别为6.47、6.78].与对照组相应时间点比较,移植组MBP、NF-H的表达量较高而GFAP的表达量较低,其中D组的MBP、NF-H的表达量最高(D组与A组MBP3周条带灰度比值为2.11,而NF-H3周条带比值为2.11),差异有统计学意义(P＜0.05),但D组GFAP各时间点的表达量差异(各周所测条带灰度比值依次为1.00:1.12:1.13:1.14)无统计学意义(P＞0.05).结论 hSCs可促进hUC-MSCs向神经元与少突胶质细胞方向分化并增强其对胶质瘢痕的抑制作用.
목적 관찰인설왕세포(hSCs)대인제대간충질간세포(hUC-MSCs)재척수손상(SCI)체내존활、분화적영향,통과분자생물학기술검측효질섬유산성단백(GFAP)、수린지감성단백(MBP)여고분자량신경사단백(NF-H)적표체,묘술기변화적시간특정.방법 분리、배양급순화hUC-MSCs화hSCs.취8주령자성Wistar대서60지,이Impactor Model-Ⅱ형타격기제작척수흉10(T10)손상모형.대서수궤균분위4조:DMEM대조조(A)、hSCs이식조(B)、hUC-MSCs이식조(C)、hUC-MSCs여hSCs연합이식조(D),각조분별우이식후1、2、3、4주취재.결과 D조가견MBP여핵인자(NF)-H염색양성세포,기타각조위음성.A조3충신경표지물재단백여mRNA수평적표체량수시간연장균축점증고,차GFAP재이식후2주명현증고,MBP여NF-H재이식후3주명현증고[2주여1주GFAP조대회도비치위1.34,이취합매련반응(PCR)소측비치위2.86；3주여1주MBP화NF-H조대회도비치분별위3.43、4.12,이PCR소측비치분별위6.47、6.78].여대조조상응시간점비교,이식조MBP、NF-H적표체량교고이GFAP적표체량교저,기중D조적MBP、NF-H적표체량최고(D조여A조MBP3주조대회도비치위2.11,이NF-H3주조대비치위2.11),차이유통계학의의(P＜0.05),단D조GFAP각시간점적표체량차이(각주소측조대회도비치의차위1.00:1.12:1.13:1.14)무통계학의의(P＞0.05).결론 hSCs가촉진hUC-MSCs향신경원여소돌효질세포방향분화병증강기대효질반흔적억제작용.
Objective To discuss the influence of human schwann cells (hSCs) on the survival and differentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs) in Wistar rats with spinal cord injury (SCI):To detect the expression of glial fibrillary acidic protein (GFAP),myelin basic protein (MBP) and high molecular weight neurofilament ( NF-H),and describe their changes of the time-dependent characteristic.Methods Firstly,the hUC-MSCs and the hSCs were isolated and purified in vitro.Sixty 8-week-old female Wistar rats were subjected to T10 SCI by Impaetor Model- Ⅱ type device and divided into four groups randomly and evenly:DMEM control group (A),hSCs transplantation group (B),hUC-MSCs transplantation group (C),co-transplantation group of hSCs and hUC-MSCs (D).The injured spinal cord tissues were taken out at 1st,2nd,3rd,and 4th week after cell transplantation.Results The MBP and NF-H positive cells could be seen in group D,but not in other groups.The three neural markers in group A were gradually increased with time,and the expression of GFAP was significantly increased at the 2nd week following transplantation,while the rest two markers were significantly increased at the 3rd week (The GFAP band gray-value ratio at 2nd week to that at 1st week was 1.34∶ 1.00,while PCR ratio was 2.86∶ 1.00; The MBP band gray-value ratio at 3rd week to that at 1st week was 3.43∶ 1.00,while PCR ratio was 6.47∶ 1.00; For NF-H,4.12∶1.00 and 6.78∶1.00 respectively).As compared with the group A at the corresponding time point,the expression of both MBP and NF-H was increased in groups B,C and D,while the expression of GFAP was reduced,more precisely,the expression of MBP and NF-H was the highest in group D (The MBP and NF-H band gray-value ratio at 3rd week to that at 1st week in both group D and group A was 2.11∶ 1.00,P＜0.05).However,there was no significant change in the GFAP expression at each time point in group D (The band gray ratio of each week was 1.00∶ 1.12∶ 1.13∶1.14) (P ＞0.05).Conclusion hUC-MSCs can differentiate into neurons and oligodendrocytes with the effect of hSCs,and its inhibitory effect on the formation of glial scar is enhanced.