目的 探讨Sox9(sex determining region Y-box9)和β-catenin对先天性马蹄内翻足(congenital talipes equinovarus,CTE)的影响以及Wnt/β-catenin信号通路作用机制。 方法 将孕10 d的SD大鼠随机均分为实验组及对照组,以135 mg/kg全反式维甲酸溶于矿物油对实验组大鼠进行灌胃制作胎鼠CTE模型,对照组予以等量矿物油灌胃处理,取大鼠足踝部组织,通过免疫组化、RT-PCR、Western blot检测β-catenin、Sox9以及磷酸化β-catenin-S552的表达水平。 结果 与对照组相比,HE染色可见CTE模型组织中有较多的胶原组织沉积,免疫组化结果显示实验组标本Sox9与β-catenin表达增高,RT-qPCR表明实验组Sox9与β-catenin的mRNA水平显著增高,Western blot结果显示实验组Sox9与β-catenin的表达增高而磷酸化β-catenin-S552表达降低。 结论 CTE大鼠的足踝部组织中Sox9高表达受Wnt/β-catenin信号通路调控,该通路参与先天性马蹄内翻足畸形的形成。
Abstract
Objective To investigate the effect of sex determining region Y-box 9 (Sox9), β-catenin and Wnt/ β-catenin signaling pathway on the congenital talipes equinovarus (CTE) and its mechanism. Methods SD rats at 10 days of gestation were randomly divided into an experimental group and a control group. All trans retinoic acid (135 mg/kg) was dissolved in mineral oil to make fetal rat CTE model in the experimental group. The control group was given the same amount of mineral oil. Immunohistochemistry, RT-PCR and Western blot were used to detect the expression levels of β-catenin, Sox9 and phosphorylated β-catenin-s552 in ankle soft tissue of control group and model group. Results Compared with the control group, HE staining showed more collagen deposition in CE model tissues, immunohistochemical results showed that the expression of β-catenin and Sox9 in the model group significantly increased. RT-qPCR result showed that the mRNA level of Sox9, β-catenin increased. Western blot result showed that the expression of P-β-catenin-s552 increased, the expression of Sox9 and β-catenin in the experimental group decreased. Conclusions The high expression of Sox9 in the ankle tissue of rats with congenital clubfoot is regulated by Wnt / β-catenin signaling pathway, which is involved in the formation of congenital clubfoot.
关键词
先天性马蹄内翻足 /
Sox9 /
Wnt信号通路 /
β-catenin /
P-β-catenin
Key words
Congenital talipes equinovarus /
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Sox9 /
Wnt signaling pathway /
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β-catenin /
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Phosphorylated-β-catenin
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参考文献
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基金
辽宁省大学生创新创业训练计划(S202010164003)