miR-145 通过靶向 Arf6 改善巨噬细胞介导的炎症反应。
MiR-145 improves macrophage-mediated inflammation through targeting Arf6.
机构信息
Department of Endocrinology, Huashan Hospital, Fudan University, Shanghai, 200040, China.
Institute of Endocrinology and Diabetology, Fudan University, Shanghai, 200040, China.
出版信息
Endocrine. 2018 Apr;60(1):73-82. doi: 10.1007/s12020-018-1521-8. Epub 2018 Jan 31.
PURPOSE
To explore the relationship between miR-145 and ADP ribosylation factor 6 (Arf6) in regulating macrophage-mediated inflammation.
METHODS
THP-1 cells were induced by 160 nM of phorbol 12myristate 13-acetate (PMA) for 48 h to differentiate to macrophages and then were treated with LPS (100 ng/ml) for 8 h to simulate chronic metabolic inflammation in vitro. Dual-luciferase reporter assay was performed. MiR-145 siRNA and LV-ARF6-RNAi were used to up or down regulate miR-145 and Arf6 expression in THP-1 cells, respectively. Omental adipose tissue from patients in surgical ward were collected to detect the expression of miR-145, Arf6 and production of proinflammatory cytokines. Patients were divided into three groups according to their body mass index and history of diabetes.
RESULTS
Dual-luciferase reporter assays showed the direct down-regulation of Arf6 by miR-145. Forty-eight-hour-transfection of miR-145 inhibitor resulted in significant increase of Arf6, IL-1beta, TNF-alpha and IL-6 as well as phosphorylation of p65 in NF-kappaB pathway in THP-1 cells, which, inversely, were reversed by overexpressing miR-145. In addition, down-regulation of Arf6 in macrophages reduced expression and secretion of cytokines. Expression of miR-145 was found to be attenuated in the omental adipose tissue of obese patients and diabetics with greater Arf6 expression, confirming the role of miR-145 in regulating macrophage-mediated inflammation targeting Arf6.
CONCLUSIONS
By means of reducing the expression of Arf6 and subsequent signal transduction via NF-kappaB, miR-145 plays a role in inhibiting the secretion of inflammatory factors and then improving the inflammatory status. MiR-145 might be one of the candidates for anti-inflammatory treatment for metabolic diseases.
目的
探讨 miR-145 与 ADP 核糖基化因子 6（Arf6）在调节巨噬细胞介导的炎症中的关系。
方法
用 160nM 的佛波醇 12-肉豆蔻酸 13-乙酸酯（PMA）诱导 THP-1 细胞 48 小时分化为巨噬细胞，然后用 LPS（100ng/ml）处理 8 小时模拟体外慢性代谢性炎症。进行双荧光素酶报告基因检测。用 miR-145 siRNA 和 LV-ARF6-RNAi 分别上调或下调 THP-1 细胞中 miR-145 和 Arf6 的表达。从外科病房的患者腹部脂肪组织中检测 miR-145、Arf6 的表达和促炎细胞因子的产生。根据患者的体重指数和糖尿病史将患者分为三组。
结果
双荧光素酶报告基因检测显示 miR-145 可直接下调 Arf6。miR-145 抑制剂转染 48 小时后，THP-1 细胞中 Arf6、IL-1β、TNF-α 和 IL-6 以及 NF-κB 通路中 p65 的磷酸化显著增加，而 miR-145 的过表达则相反。此外，巨噬细胞中 Arf6 的下调减少了细胞因子的表达和分泌。在肥胖患者和糖尿病患者的大网膜脂肪组织中发现 miR-145 的表达减弱，Arf6 表达增加，证实了 miR-145 通过靶向 Arf6 调节巨噬细胞介导的炎症的作用。
结论
通过降低 Arf6 的表达及其随后通过 NF-κB 的信号转导，miR-145 在抑制炎症因子的分泌从而改善炎症状态中发挥作用。miR-145 可能是代谢性疾病抗炎治疗的候选药物之一。
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