CAJ | 학술논문

目的评价缺氧预处理骨髓间充质干细胞(BMSC)对大鼠脊髓缺血再灌注时血红素氧合酶-1(HO-1)表达的影响.方法健康清洁级SD雄性大鼠30只,6～8周龄,体重200～ 250 g,采用随机数字表法,将其分为5组(n=6):假手术组(S组)、缺血再灌注组(I/R组)、PBS组、BMSC移植组(BMSC组)和缺氧预处理组(HP).HP组BMSC置于3％O2-5％ CO2-92％N2的培养箱中培养24 h,进行缺氧预处理.PBS组、BMSC组和HP组分别鞘内注射PBS、BMSC和缺氧预处理的BMSC,60 min后采用主动脉阻断法建立大鼠脊髓缺血再灌注损伤模型.分别于再灌注6、12、24、48 h和7d时行神经功能评分.于再灌注7d时,处死大鼠,取脊髓组织,采用Western blot法测定HO-1蛋白表达水平,采用RT-PCR法测定HO-1 mRNA表达水平.结果与S组比较,其余各组再灌注各时点神经功能评分降低,脊髓组织HO-1 mRNA及其蛋白表达上调(P＜0.05);与I/R组比较,BMSC组和HP组再灌注各时点神经功能评分升高,脊髓组织HO-1 mRNA及其蛋白表达上调(P＜0.05),PBS组再灌注各时点神经功能评分、脊髓组织HO-1 mRNA及其蛋白表达差异无统计学意义(P＞0.05);与BMSC组比较,HP组再灌注各时点神经功能评分升高,脊髓组织HO-1 mRNA及其蛋白表达上调(P＜0.05).结论缺氧预处理增强BMSC移植减轻大鼠脊髓缺血再灌注损伤作用的机制可能与HO-1表达进一步上调有关.
목적 평개결양예처리골수간충질간세포(BMSC)대대서척수결혈재관주시혈홍소양합매-1(HO-1)표체적영향.방법 건강청길급SD웅성대서30지,6～8주령,체중200～ 250 g,채용수궤수자표법,장기분위5조(n=6):가수술조(S조)、결혈재관주조(I/R조)、PBS조、BMSC이식조(BMSC조)화결양예처리조(HP).HP조BMSC치우3％O2-5％ CO2-92％N2적배양상중배양24 h,진행결양예처리.PBS조、BMSC조화HP조분별초내주사PBS、BMSC화결양예처리적BMSC,60 min후채용주동맥조단법건립대서척수결혈재관주손상모형.분별우재관주6、12、24、48 h화7d시행신경공능평분.우재관주7d시,처사대서,취척수조직,채용Western blot법측정HO-1단백표체수평,채용RT-PCR법측정HO-1 mRNA표체수평.결과 여S조비교,기여각조재관주각시점신경공능평분강저,척수조직HO-1 mRNA급기단백표체상조(P＜0.05);여I/R조비교,BMSC조화HP조재관주각시점신경공능평분승고,척수조직HO-1 mRNA급기단백표체상조(P＜0.05),PBS조재관주각시점신경공능평분、척수조직HO-1 mRNA급기단백표체차이무통계학의의(P＞0.05);여BMSC조비교,HP조재관주각시점신경공능평분승고,척수조직HO-1 mRNA급기단백표체상조(P＜0.05).결론 결양예처리증강BMSC이식감경대서척수결혈재관주손상작용적궤제가능여HO-1표체진일보상조유관.
Objective To evaluate the effect of hypoxia-preconditioned bone marrow mesenchymal stem cells (BMSCs) on the expression of HO-1 during spinal ischemia-reperfusion (I/R) in rats.Methods Thirty healthy adult Sprague-Dawley rats,weighing 200-250 g,were randomly divided into 5 groups with 6 animals in each group using a random number table:sham operation group (group S),I/R group,PBS group,BMSC transplantation group (BMSC group) and hypoxic preconditioning group (group HP).In group HP,BMSCs were exposed to 3％O2-5％CO2-92％N2 for 24 h for hypoxia preconditioning.In PBS,BMSC and HP groups,PBS,BMSCs and BMSCs for hypoxic preconditioning were injected intrathecally,respectively,and 60 min later spinal I/R was induced by clamping the aorta in the rats anesthetized with chloral hydrate.At 6,12,24 and 48 h and 7 days of reperfusion,the neurological function was evaluated and scored.At day 7 of reperfusion,the rats were sacrificed,and spinal cord tissues were obtained for determination of HO-1 expression (by Western blot) and HO-1 mRNA expression (using real-time PCR).Results Compared with group S,the neurological function score was significantly decreased at each time point of reperfusion,and the expression of HO-1 mRNA and protein in spinal cord tissues was up-regulated in the other groups.Compared with group I/R,the neurological function score was significantly increased at each time point of reperfusion,and the expression of HO-1 mRNA and protein in spinal cord tissues was upregulated in BMSC and HP groups,and no significant change was found in group PBS in the parameters mentioned above.Compared with group BMSC,the neurological function score was significantly increased at each time point of reperfusion,and the expression of HO-1 mRNA and protein in spinal cord tissues was up-regulated in group HP.Conclusion The mechanism by which hypoxic preconditioning enhances protection of spinal cord by BMSC transplantation may be related to up-regulation of HO-1 expression in rats.