| 摘要: |
| 目的探讨低氧预处理大鼠骨髓间充质干细胞（BMSCs）移植对大鼠缺血-再灌注（I/R）急性肾损伤（AKI）的修复作用及其可能机制。方法采用200滋mol/L氯化钴（CoCl2）低氧预处理大鼠BMSCs。夹闭双侧肾蒂40min建立大鼠I/RAKI模型，按随机数字表法分为颈动脉注射细胞培养基组（control组）、常氧培养BMSCs移植组（normoxia+BMSCs组）及低氧预处理BMSCs移植组（Co+BMSCs组），每组24只。BMSCs移植再灌注后24、48、72h及1周时每组各处死大鼠6只，采用开胸心腔内采血方法采集血样检测血清尿素氮（BUN）和血肌酐（Scr）水平，取肾脏组织观察形态学变化和肾小管间质损伤程度并评分；采用ELISA法和RT-PCR法检测再灌注后24h大鼠肾脏组织中血管内皮生长因子（VEGF）、碱性成纤维母细胞生长因子（bFGF）、肝细胞生长因子（HGF）、胰岛素样生长因子-1（IGF-1）的表达水平。结果(1)再灌注后24h，Co+BMSCs组、normoxia+BMSCs组大鼠BUN、Scr水平均低于control组（均P＜0.05），且Co+BMSCs组低于normoxia+BMSCs组（P＜0.05）；肾小管间质损伤程度以control组最重，normoxia+BMSCs组次之，Co+BMSCs组最轻；再灌注后48h，Co+BMSCs组大鼠肾小管上皮细胞修复最明显，normoxia+BMSCs组次之；再灌注后1周，Co+BMSCs组大鼠BUN、Scr水平仍低于normoxia+BMSCs组和control组（均P＜0.05），normoxia+BMSCs组和control组出现了明显的肾小管上皮细胞萎缩及间质纤维化等慢性改变，而Co+BMSCs组无此改变。（2）再灌注后24h，normoxia+BMSCs组大鼠bFGF表达水平高于control组（P＜0.05），而Co+BMSCs组的bFGF表达水平明显高于normoxia+BMSCs组（P＜0.05）；normoxia+BMSCs组大鼠HGF表达水平高于control组，但差异无统计学意义（P＞0.05），而Co+BMSCs组的bFGF表达水平均高于normoxia+BMSCs组和control组（均P＜0.05）；3组大鼠VEGF和IGF-1表达水平比较差异均无统计学意义（均P＞0.05）。结论与常氧培养BMSCs移植相比，低氧预处理后的BMSCs移植能更有效改善I/R大鼠肾功能、修复肾脏组织损伤，其作用机制可能与低氧预处理能更有效地促进BMSCs旁分泌HGF和bFGF等有关。 |
| 关键词: 骨髓间充质干细胞 低氧 急性肾损伤 旁分泌 |
| DOI： |
| 分类号: |
| 基金项目:国家自然科学基金；上海青年医师培养资助计划；复旦大学附属中山医院优秀青年计划 |
|
| Effects of hypoxia-preconditioning of bone marrow derived mesenchymal stem cells on rat acute kidney injury |
|
LI Hong, DING Xiaoqiang, WANG Hua, CHEN Hongyu, YU Xiaofang
|
|
Hangzhou Hospital of Traditional Chinese Medicine
|
| Abstract: |
| Objective To investigate the effects and mechanism of hypoxia-preconditioning of bone marrow derived mesenchymal stem cells(BMSCs) on acute ischemia/reperfusion(I/R) injury of rat kidney. Methods Hypoxia-preconditioning was performed on BMSCs by treatment with 200滋mol/L CoCl2 for 24h. I/R model was induced by cross-clasped for 40min followed by reperfusion in 72 SD rats and the animals were divided into 3 groups with 24 in each group. Rats in control group received carotid injection of cell culture medium, rats in normoxic BMSCs group received carotid injection of normoxia-cultured BMSCs and rats in hypoxic BMSCs group received carotid injection of hypoxia-preconditioned BMSCs. The renal function was estimated by measurement of serum creatinine (Scr) and blood urea nitrogen (BUN), and histological changes were observed by hematoxylin
and eosin (HE) staining at 24h, 48h, 72h and 1wk after injection. The protein and mRNA expressions of VEGF, bFGF, HGF and IGF-1 in kidney were detected by ELISA and real-time PCR, respectively. Results At 24h after reperfusion, the BUN and Scr levels in both two BMSCs groups were significantly lower than those in control group (P＜0.05), and the levels in hypoxic BMSCs group was even lower than those in normoxic BMSCs group(P＜0.05), the renal tubulointerstitial damage of hypoxic BMSCs group was significantly lower than that of the normoxic BMSCs group and the control group (P＜0.05). at 48h after reperfusion, the renal tubular epithelial cells of the hypoxic BMSCs group recovered earlier than other two groups at 1 week after reperfusion, the BUN and Scr levels of hypoxic BMSCs group was still significantly lower than those of other two groups (P＜0.05), and there were no chronic changes in the hypoxic BMSCs group. The protein and mRNA levels of HGF and bFGF of hypoxic BMSCs group were significantly higher than those of the other two groups (P＜0.05), and the levels of HGF and bFGF in normoxic BMSCs group was higher than those in the control group. Conclusion Infusion of hypoxia-preconditioned BMSCs can enhance the recovery of IR injure of rat kidney, which may be attributable to the up-regulation of paracrine factors of HGF and bFGF after hypoxia precondi- tioning in BMSCs. |
| Key words: Mesenchymal stem cells Hypoxia Acute kidney injury Paracrine |
