MicroRNA-335-5p Plays Dual Roles in Periapical Lesions by Complex Regulation Pathways
MicroRNA-335-5p Plays Dual Roles in Periapical Lesions by Complex Regulation Pathways
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MicroRNA-335-5p 通过复杂的调控途径在根尖周病变中发挥双重作用
DOI：
10.1016/j.joen.2017.03.018
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发表时间：
2017
影响因子：
4.2
通讯作者：
Huang Dingming
中科院分区：
文献类型：
--
作者：
Yue Junli;Wang Puyu;Hong Qingchun;Liao Qian;Yan Li;Xu Weizhe;Chen Xi;Zheng Qinghua;Zhang Lan;Huang Dingming
MicroRNA-335- 5 p可调控骨髓间充质干细胞向成骨细胞和软骨细胞分化。为探讨miR-335- 5 p在根尖周炎（AP）中的作用及其调控机制，采用裂解液法提取人牙周膜成纤维细胞（HPDLFs）、10例AP组织和6例健康牙周膜组织的总RNA。采用实时定量聚合酶链反应检测基因表达。使用双重荧光素酶测定（Promega，麦迪逊，WI）来测试miR-335- 5 p直接靶向的尿激酶型纤溶酶原激活物受体（uPAR）和核因子κ-B配体的受体激活物（RANKL）。Western Blot检测RANKL、uPAR和脆性X相关基因1（FXR 1）的蛋白表达。采用酶联免疫吸附试验检测白细胞介素6、肿瘤坏死因子α和RANKL的分泌。结果miR-335- 5 p在HPDLF炎症中起阳性介导作用（P<0.05）。miR-335- 5 p的两个靶标uPAR和RANKL被鉴定。有趣的是，uPAR在基础水平上被miR-335- 5 p抑制，但当HPDLF受到脂多糖刺激时，它可以从miR-335- 5 p介导的抑制中解脱出来，这被称为去抑制。miR-335- 5 p促进HPDLFs中的RANKL，无论其是否处于炎症条件下（P< .05）。我们证明FXR 1负责miR-335- 5 p对uPAR的去抑制（P< .01）。FXR 1和uPAR均为HPDLF炎症的阳性介质（P< .05）。miR-335- 5 p、uPAR、RANKL和FXR 1在HPDLF炎症组织和AP组织中的表达差异无统计学意义（P> 0.05）。
IntroductionMicroRNA-335-5p has been reported to regulate osteogenic and chondrogenic differentiations of mesenchymal stem cells. The aim of this study was to explore the function and regulation mechanism of miR-335-5p in apical periodontitis (AP).MethodsTotal RNAs were extracted from human periodontal ligament fibroblasts (HPDLFs), 10 AP tissues, and 6 healthy periodontal ligament tissues using lysis buffer. Gene expression was detected using real-time polymerase chain reaction. The Dual Luciferase Assay (Promega, Madison, WI) was used to test miR-335-5p directly targeted urokinase-type plasminogen activator receptor (uPAR) and the receptor activator of nuclear factor kappa-B ligand (RANKL). Western Blot was used to detect protein expressions of RANKL, uPAR, and the fragile X–related 1 gene (FXR1). The enzyme-linked immunosorbent assay was used to detect the secretions of interleukin 6, tumor necrosis factor alpha, and RANKL. Data were analyzed using the Studentttest.ResultsmiR-335-5p acted as a positive mediator in HPDLF inflammation (P< .05). Two targets of miR-335-5p, uPAR and RANKL, were identified. Interestingly, uPAR was repressed by miR-335-5p at the basal level, but it can be relieved from miR-335-5p–mediated repression, which is called derepression, when HPDLFs were subjected to lipopolysaccharide stimulation. miR-335-5p promoted RANKL in HPDLFs regardless of whether or not it was under inflammatory conditions (P< .05). We proved FXR1 was responsible for the derepression of uPAR from miR-335-5p (P< .01). Both FXR1 and uPAR were positive mediators in HPDLF inflammation (P< .05). miR-335-5p, uPAR, RANKL, and FXR1 had the same expression profiles in HPDLF inflammation and AP tissues (P< .05).ConclusionsOur data showed that miR-335-5p may play dual roles in AP, and it might be considered as a target for therapeutic potency in clinical applications.