Characterization of a Novel Model of Lumbar Ligamentum Flavum Hypertrophy in Bipedal Standing Mice.
Characterization of a Novel Model of Lumbar Ligamentum Flavum Hypertrophy in Bipedal Standing Mice.
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双足站立小鼠腰椎黄韧带肥大新模型的表征
DOI：
10.1111/os.13156
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发表时间：
2021-12
影响因子：
2.1
通讯作者：
Wang L
中科院分区：
文献类型：
--
作者：
Zheng ZY;Li P;Ao X;Qian L;Peng YX;Chu J;Jiang T;Lian ZN;Zhang ZM;Wang L
探讨黄韧带肥大（HLF）发生的主要原因，并探讨采用小鼠双足站立模型模拟人黄韧带肥大病理变化的可能性。32只8周龄C57BL/6雄性小鼠随机分为实验组（n = 16）和对照组（n = 16）。实验组小鼠的恐水症诱导小鼠采取两足站立姿势。实验小鼠每天保持两足站立8小时，间隔2小时进食和饮水。对照组小鼠被放置在类似的环境中，没有两足站立。我们比较了8只18月龄C57BL/6雄性小鼠，以评估衰老因素导致的LF变性。采用三维重建和有限元模型，分析了小鼠下半关节在伸直和两足站立时的应力应变分布。采用苏木精和伊红（HE）、Verhoeff - Van Gieson （VVG）和免疫组化（IHC）染色评价小鼠和人的LF变性。采用RT - qPCR和免疫荧光分析评估COL1A1、COL3A1、α‐SMA、MMP2、IL‐1β和COX‐2的纤维化相关因子和炎症因子的表达。两足站立小鼠的von Mises应力（8.85 × 10−2 MPa）和最大主应变（6.64 × 10−1）分别增加了4944倍和7703倍。HE染色显示，10周龄双足站立组小鼠LF面积（[10.01±2.93]× 104 μm2）大于对照组（[3.76±1.87]× 104 μm2）和18月龄组（[6.09±2.70]× 104 μm2）。VVG染色显示，小鼠（3.23±0.58）和人（2.23±0.31）的HLF弹性纤维损失相似，胶原纤维增加。小鼠（39.63±4.81）和人（23.25±2.05）在HLF过程中细胞密度较高。免疫组化染色显示，小鼠（1.63±0.74）和人（3.50±1.85）HLF中α‐SMA阳性细胞数量显著增加。两足站立组炎症因子和纤维化相关因子COL1A1、COL3A1、α‐SMA、MMP2、IL‐1β、COX‐2的表达均高于对照组。我们的研究表明，三维有限元模型可以帮助分析建模小鼠LF的异常应力和应变分布。与老化相比，机械应力是导致黄韧带肥大的主要原因。两足站立小鼠模型能较好地反映人HLF的病理特征。双足站立小鼠模型可为阐明机械应力诱导HLF的分子机制提供标准化条件。小鼠的恐水症诱导其采取两足站立姿势。采用有限元方法分析了不同体位下黄韧带（LF）的应力应变分布。双足站立小鼠的von Mises应力和Maximum Principal strain分别增加了4944倍和7703倍。我们的研究表明，与衰老相比，机械应力是导致HLF的主要原因，可以用两足站立小鼠来研究HLF的体内机制。
To explore the main causes of hypertrophied ligamentum flavum (HLF) and the possibility of using bipedal standing mouse model to simulate the pathological changes in human HLF. Thirty‐two 8‐week‐old C57BL/6 male mice were randomly assigned to the experimental group (n = 16) and control group (n = 16). In the experimental group, mice were induced to adopt a bipedal standing posture by their hydrophobia. The experimental mice were maintained bipedal standing for 8 h a day with an interval of 2 h to consume food and water. The control mice were placed in a similar environment without bipedal standing. Eight 18‐month‐old C57BL/6 male mice were compared to evaluate the LF degeneration due to aging factor. Three‐dimensional (3D) reconstruction and finite element models were carried out to analyze the stress and strain distribution of the mouse LF in sprawling and bipedal standing postures. Hematoxylin and Eosin (HE), Verhoeff‐Van Gieson (VVG), and immunohistochemistry (IHC) staining were used to evaluate the LF degeneration of mice and humans. RT‐qPCR and immunofluorescence analysis were used to evaluate the expressions of fibrosis‐related factors and inflammatory cytokines of COL1A1, COL3A1, α‐SMA, MMP2, IL‐1β, and COX‐2. The von Mises stress (8.85 × 10−2 MPa) and maximum principal strain (6.64 × 10−1) in LF were increased 4944 and 7703 times, respectively, in bipedal standing mice. HE staining showed that the mouse LF area was greater in the bipedal standing 10‐week‐old group ([10.01 ± 2.93] × 104 μm2) than that in the control group ([3.76 ± 1.87] × 104 μm2) and 18‐month‐old aged group ([6.09 ± 2.70] × 104 μm2). VVG staining showed that the HLF of mice (3.23 ± 0.58) and humans (2.23 ± 0.31) had a similar loss of elastic fibers and an increase in collagen fibers. The cell density was higher during the process of HLF in mice (39.63 ± 4.81) and humans (23.25 ± 2.05). IHC staining showed that the number of α‐SMA positive cells were significantly increased in HLF of mice (1.63 ± 0.74) and humans (3.50 ± 1.85). The expressions of inflammatory cytokines and fibrosis‐related factors of COL1A1, COL3A1, α‐SMA, MMP2, IL‐1β, and COX‐2 were consistently higher in bipedal standing group than the control group. Our study suggests that 3D finite element models can help analyze the abnormal stress and strain distributions of LF in modeling mice. Mechanical stress is the main cause of hypertrophied ligamentum flavum compared to aging. The bipedal standing mice model can reflect the pathological characteristics of human HLF. The bipedal standing mice model can provide a standardized condition to elucidate the molecular mechanisms of mechanical stress‐induced HLF in vivo. Mice were induced to adopt a bipedal standing posture by their hydrophobia. Finite element analysis was carried out to analyze the stress and strain distribution of the ligamentum flavum (LF) in different postures. The von Mises stress and Maximum Principal strain in LF were increased 4944 and 7703 times, respectively, in bipedal standing mice. Our study suggests that mechanical stress is the main cause of HLF compared to aging, and that bipedal standing mice can be used to study the mechanism of HLF in vivo.
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影响因子：
4.1
作者：
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通讯作者：
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影响因子：
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影响因子：
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影响因子：
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作者：
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通讯作者：
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影响因子：
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作者：
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