CAJ | 학술논문

目的观察骨髓间充质干细胞(BMSCs)种植于丝素蛋白多孔支架(PSFSs)后对大鼠脊髓损伤(SCI)的治疗作用.方法将96只SD大鼠随机分成对照组、PSFSs组、BMSCs组及PSFSs±BMSCs移植组,每组24只.移植后l、2、4周采用Basso-Beattie-Bresnahan(BBB)评分法评估大鼠神经功能恢复情况,采用免疫组织化学染色、Western blot分析检测损伤部位生长相关蛋白43(GAP-43)、髓鞘碱性蛋白(MBP)的表达水平,免疫荧光染色检测GAP-43的表达强度.结果术后4周BBB评分实验组较对照组显著提高,各组间差异有统计学意义(分别为5.23±1.13、7.17 ±1.11、7.35±1.59、9.43 ±2.16,P＜0.05).D组GAP43免疫组织化学染色积分吸光度(IA)值高于其他组(分别为13.94±3.63、26.07±5.31、22.14±4.56、33.03±7.58),MBP免疫组织化学染色IA值高于其他组,分别为13.80±3.41、21.16±3.36、32.96±7.74、44.06 ±7.99,各组间差异有统计学意义(P＜0.05).免疫荧光染色示D组损伤区内具有更多表达GAP-43的生长锥.Western blot分析表明各组GAP43的灰度值比值分别为0.16±0.04、0.41 ±0.05、0.33 ±0.06、0.72 ±0.16,MBP的灰度值比值分别为0.96 ±0.11、1.85±0.08、2.14 ±0.16、2.43 ±0.31,差异有统计学意义(P＜0.05).结论种植BMSCs的PSFSs能够促进SCI大鼠轴突再生、髓鞘化及后肢运动功能的恢复,其疗效优于单一方法.
목적 관찰골수간충질간세포(BMSCs)충식우사소단백다공지가(PSFSs)후대대서척수손상(SCI)적치료작용.방법 장96지SD대서수궤분성대조조、PSFSs조、BMSCs조급PSFSs±BMSCs이식조,매조24지.이식후l、2、4주채용Basso-Beattie-Bresnahan(BBB)평분법평고대서신경공능회복정황,채용면역조직화학염색、Western blot분석검측손상부위생장상관단백43(GAP-43)、수초감성단백(MBP)적표체수평,면역형광염색검측GAP-43적표체강도.결과 술후4주BBB평분실험조교대조조현저제고,각조간차이유통계학의의(분별위5.23±1.13、7.17 ±1.11、7.35±1.59、9.43 ±2.16,P＜0.05).D조GAP43면역조직화학염색적분흡광도(IA)치고우기타조(분별위13.94±3.63、26.07±5.31、22.14±4.56、33.03±7.58),MBP면역조직화학염색IA치고우기타조,분별위13.80±3.41、21.16±3.36、32.96±7.74、44.06 ±7.99,각조간차이유통계학의의(P＜0.05).면역형광염색시D조손상구내구유경다표체GAP-43적생장추.Western blot분석표명각조GAP43적회도치비치분별위0.16±0.04、0.41 ±0.05、0.33 ±0.06、0.72 ±0.16,MBP적회도치비치분별위0.96 ±0.11、1.85±0.08、2.14 ±0.16、2.43 ±0.31,차이유통계학의의(P＜0.05).결론 충식BMSCs적PSFSs능구촉진SCI대서축돌재생、수초화급후지운동공능적회복,기료효우우단일방법.
Objective To investigate the therapeutic effects of porous silk fibroin scaffolds (PSFSs) seeded with bone mesenchymal stem cells (BMSCs) on the repair of transversal spinal cord injury (SCI) in rats.Methods Ninety-six SD rats were randomly divided into control group,PSFSs group,BMSCs group and PSFSs plus BMSCs group (24 each).The function repair was evaluated by Basso-Beattie-Bresnahan (BBB) score system at lst,2nd and 4th week.Immunohistochemistry and Western blotting were applied to detect the expression of growth associated protein 43 (GAP-43) and myelin basic protein (MBP) in the injured spinal cord.The intensity of GAP-43 expression was measured by immuno-fluorescence staining.Results At 4th week after injury,the implanted groups exhibited a more significant improvement in hindlimb locomotor function than in control group in BBB scores (P ＜0.05).The BBB sores in control group,PSFSs group,BMSCs group and PSFSs plus BMSCs group were 5.23 ± 1.13,7.17 ±1.11,7.35 ± 1.59 and 9.43 ± 2.16 respectively.Immunohistochemistry showed that the integral absorbance (IA) values of GAP-43 and MBP in implanted groups were significantly higher than those in control group (P ＜ 0.05).In control group,PSFSs group,BMSCs group and PSFSs plus BMSCs group,the IA values of GAP-43 were 13.94 ±3.63,26.07 ±5.31,22.14 ±4.56 and 33.03 ±7.58,and those of MBP were 13.80 ± 3.41,21.16 ± 3.36,32.96 ± 7.74 and 44.06 ± 7.99,respectively.The immuno-fluorescence staining indicated that there were more GAP-43 positive growth cones at the lesion site in PSFSs plus BMSCs group.Western blotting results showed that there was significant difference in the expression of GAP-43 and MBP proteins between control group and implanted groups (P ＜ 0.05).In control group,PSFSs group,BMSCs group and PSFSs plus BMSCs group,the gray value ratio of GAP-432 was 0.16 ± 0.04,0.41 ±0.05,0.33 ±0.06 and 0.72 ±0.16,and that of MBP was 0.96 ±0.11,1.85 ±0.08,2.14 ±0.16 and 2.43 ± 0.31,respectively.Conclusion Transplantation of PSFSs seeded with BMSCs can promote the axonal regeneration,myelination and functional recovery after SCI in rats.