The role of interleukin-6/interleukin-6 receptor signaling in the mechanical stress-induced extracellular matrix remodeling of bladder smooth muscle
The role of interleukin-6/interleukin-6 receptor signaling in the mechanical stress-induced extracellular matrix remodeling of bladder smooth muscle
复制标题
IL-6/IL-6受体信号在机械应力诱导的膀胱平滑肌细胞外基质重塑中的作用
DOI：
10.1016/j.abb.2020.108674
复制
发表时间：
2021-03-31
影响因子：
3.9
通讯作者：
Wang, Kunjie
中科院分区：
文献类型：
--
作者：
He, Qing;Lin, Yifei;Wang, Kunjie
细胞外基质（ECM）重塑与膀胱出口梗阻（BOO）引起的病理变化密切相关。在这项研究中，我们研究了白细胞介素-6（IL-6）在机械牵张诱导的膀胱平滑肌ECM重塑中的作用。为了建立BOO动物模型，雌性Sprague？将道利大鼠部分结扎。此外，增加静水压力和机械拉伸施加到人膀胱平滑肌细胞（HBSMCs）作为体外模型。使用DNA微阵列分析大鼠炎症基因的表达。采用定量RT-PCR和免疫组织化学方法检测大鼠膀胱平滑肌中IL-6的表达。为了确定IL-6的特异性，将小干扰核糖核酸（siRNA）转染和IL-6受体抑制剂（SC 144）应用于HBSMCs。还使用siRNA转染的qRT-PCR来确定下游信号传导的特异性。此外，进行蛋白质印迹以验证表达结果。在动物模型中，ECM组分和炎症基因的表达显著上调。BOO大鼠IL-6在mRNA水平和蛋白水平的表达均增加。在体外，静水压力，和机械拉伸促进MMP 7和MMP 11的表达。此外，在静水压力组和机械拉伸组中均出现III型胶原的下调。然而，纤维连接蛋白的表达表现出相反的模式之间的静水压力和机械拉伸组。应用靶向siRNA转染和靶向IL-6的抑制剂（SC 144）显著逆转了机械应力下MMP 7和MMP 11的变化，并部分增加了III型胶原和纤连蛋白的表达。综上所述，IL-6参与了机械应力下HBSMCs的ECM重塑，提示IL-6可能在BOO中起重要作用。
Extracellular matrix (ECM) remodeling is strongly associated with pathological changes induced by bladder outlet obstruction (BOO). In this study, we investigated the role of interleukin-6 (IL-6) in mechanical stretchinduced ECM remodeling of bladder smooth muscle. To construct a BOO animal model, the urethras of female Sprague?Dawley rats were partially ligated. In addition, increased hydrostatic pressure and mechanical stretching were applied to human bladder smooth muscle cells (HBSMCs) as an in vitro model. The expression of rat inflammatory genes was analyzed using DNA microarrays. We used quantitative RT-PCR (qRT-PCR) and immunohistochemical staining to detect IL-6 in the bladder smooth muscle of rats. To determine the specificity of IL-6, small interfering ribonucleic acid (siRNA) transfection and IL-6 receptor inhibitor (SC144) were applied to HBSMCs. qRT-PCR with siRNA transfection was also used to determine the specificity of downstream signaling. Moreover, western blotting was conducted to verify the expression results. In the animal model, the expression of ECM components and inflammatory genes was significantly upregulated. The expression of IL-6 was increased at both the mRNA level and the protein level in BOO rats. In vitro, hydrostatic pressure, and mechanical stretching both promoted MMP7 and MMP11 expression. Additionally, downregulation of collagen III occurred in both the hydrostatic pressure group and the mechanical stretch group. However, the expression of fibronectin exhibited opposing patterns between the hydrostatic pressure and mechanical stretch groups. The application of targeted siRNA transfection and an inhibitor (SC144) that targeted IL-6 significantly reversed the changes in MMP7 and MMP11 under mechanical stress and partially increased the expression of collagen III and fibronectin. In summary, IL-6 participated in the ECM remodeling of HBSMCs under mechanical stress, indicating that IL-6 may play an essential role in BOO.