High cholesterol inhibits tendon-related gene expressions in tendon-derived stem cells through reactive oxygen species-activated nuclear factor-κB signaling
High cholesterol inhibits tendon-related gene expressions in tendon-derived stem cells through reactive oxygen species-activated nuclear factor-κB signaling
复制标题
高胆固醇通过活性氧激活的核因子-κB 信号传导抑制肌腱干细胞中肌腱相关基因的表达。
DOI：
10.1002/jcp.28433
复制
发表时间：
2019-10-01
影响因子：
5.6
通讯作者：
Zhang, Kairui
中科院分区：
文献类型：
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作者：
Li, Kaiqun;Deng, Ganming;Zhang, Kairui
临床研究表明，血清胆固醇水平升高会增加高胆固醇血症患者发生肌腱病变的风险，但胆固醇对肌腱源性干细胞（TDSCs）的影响及其潜在机制尚未研究。本研究的目的是在体外和体内研究胆固醇与肌腱病变之间的关系，并探讨其潜在的分子机制。在TDSC中，胆固醇的作用通过定量聚合酶链反应、蛋白质印迹分析和免疫荧光染色来评估。采用流式细胞术检测细胞内活性氧（ROS）水平。使用代表性I κ B激酶（IKK）抑制剂BAY 11-7082评价核因子（NF）-κ B信号传导与胆固醇作用之间的联系。此外，使用苏木精和伊红染色和免疫组织化学对高脂饮食喂养的载脂蛋白E小鼠的跟腱进行组织学评估。我们发现，高胆固醇明显降低表达的肌腱细胞标志物（胶原1，硬化，tenomodulin），并通过NF-κ B B途径在体外和体内升高ROS水平。ROS清除剂N-乙酰半胱氨酸（NAC）和BAY 11-7082可逆转胆固醇对TDSC肌腱相关基因表达的抑制作用。此外，NAC阻断胆固醇诱导的I κ B α和p65磷酸化。高胆固醇血症组跟腱组织学改变明显。这些结果表明，高胆固醇可能通过ROS激活的NF-κ B信号通路抑制TDSC中肌腱相关基因的表达，提示高胆固醇血症中肌腱病变的发病机制，并提示高胆固醇血症诱导肌腱病变的新机制。
Clinical studies have indicated that increased serum cholesterol levels raised the risk of tendinopathy in hypercholesterolemia, but the effect of cholesterol on tendon-derived stem cells (TDSCs) and its underlying mechanism have not been studied. The purpose of this study is to investigate the association between cholesterol and tendinopathy in vitro and in vivo, and its underlying molecular mechanism as well. In TDSCs, the effect of cholesterol was assessed by quantitative polymerase chain reaction, western blot analysis, and immunofluorescence staining. Intracellular levels of reactive oxygen species (ROS) was detected, using flow cytometry. The link between nuclear factor (NF)-kappa B signaling and the effect of cholesterol was evaluated using a representative I kappa B kinase (IKK) inhibitor, BAY 11-7082. In addition, Achilles tendons from apolipoprotein E mice fed with a high-fat diet were histologically assessed using hematoxylin and eosin staining and immunohistochemistry. We found that high cholesterol apparently lowered the expression of tendon cell markers (collagen 1, scleraxis, tenomodulin), and elevated ROS levels via the NF-kappa B pathway both in vitro and in vivo. The ROS scavenger N-acetylcysteine (NAC) and BAY 11-7082 reversed the inhibiting effect of cholesterol on the tendon-related gene expressions of TDSCs. Moreover, NAC blocked cholesterol-induced phosphorylation of I kappa B alpha and p65. Significant histological alternation in vivo was shown in Achilles tendon in the hypercholesterolemic group. These results indicated that high cholesterol may inhibit the tendon-related gene expressions in TDSCs via ROS-activated NF-kB signaling, implying pathogenesis of tendinopathy in hypercholesterolemia and suggesting a new mechanism underlying hypercholesterolemia-induced tendinopathy.