Increased Expression of Y-Box-Binding Protein-1 in Hind-Limb Muscles During Regeneration from Ischemic Injury in Mice
Increased Expression of Y-Box-Binding Protein-1 in Hind-Limb Muscles During Regeneration from Ischemic Injury in Mice
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DOI：
10.1620/tjem.244.53
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发表时间：
2018-01-01
影响因子：
2.2
通讯作者：
Ito, Ken-ichi
中科院分区：
文献类型：
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作者：
Fuke, Megumi;Narita, Makoto;Ito, Ken-ichi
严重肢体缺血(CLI)是外周动脉疾病(PAD)最严重的并发症。了解CLI后组织修复的分子机制对于预防PAD进展是必要的。Y-box结合蛋白-1(YB-1)调节多种基因的表达以响应环境胁迫。我们的目的是确定YB-1是否参与了缺血肌肉的再生。建立小鼠后肢缺血模型，即结扎左后肢的股动脉、隐动脉和月国动脉。右后肢，仅有皮肤伤口，作为对照。分别于术前0d、术后1d、2d、7d、10d、14d检查后肢(n=3~5)，取股二头肌、内收肌、股直肌、股薄肌进行组织病理学和免疫组织化学分析。在缺血肢体中，由肌管增加引发的肌肉生成从第7天开始；此后，再生肌肉的体积逐渐增加。RT-PCR分析显示，肢体缺血损伤后YB-1mRNA表达水平升高，第2天达高峰，随后下降。第7天，缺血肌MyoD和α-SMA的mRNAs表达水平显著高于对照组。免疫组织化学分析显示，7d成肌细胞和肌管YB-1免疫反应增强，14d下降，肌管内α-SMA和肌球蛋白重链一过性增加。这是首次报道在缺血损伤后肌肉再生过程中YB-1的表达增加。
Critical limb ischemia (CLI) is the most severe complication of peripheral arterial disease (PAD). Understanding the molecular mechanisms underlying tissue repair after CLI is necessary for preventing PAD progression. Y-box binding protein-1 (YB-1) regulates the expression of many genes in response to environmental stresses. We aimed to determine whether YB-1 is involved in ischemic muscle regeneration. A mouse ischemic hind-limb model was generated; namely, the femoral, saphenous, and popliteal arteries in the left hind limb were ligated. The right hind limb, with skin incisions alone, served as control. Hind limbs (n = 3-5 for each time point) were examined on day 0 (before the operation) and on postoperative days 1, 2, 7, 10, and 14, and the biceps femoris, adductor, rectus femoris, and gracilis muscles were subjected to histopathological and immunohistochemical analyses. In ischemic limbs, myogenesis, triggered by an increase in myotubes, began on day 7; thereafter, regenerated muscles gradually increased in volume. RT-PCR analysis showed that YB-1 mRNA levels were increased in the limbs after ischemic injury, peaked on day 2, and subsequently decreased. On day 7, expression levels of MyoD and alpha smooth muscle actin (alpha SMA) mRNAs were significantly higher in ischemic muscles than in control muscles. Immunohistochemical analysis revealed increased YB-1 immunoreactivity in myoblasts and myotubes on day 7, which was decreased by day 14. The immunoreactive alpha SMA and smooth muscle myosin heavy chain were transiently increased in myotubes. This is the first report showing the increased expression of YB-1 during muscle regeneration after ischemic injury.