Hu Haixia, Zhu Xiaoqin, Lin Xinjun
Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, 1 Qiuyang Road, Minhou Shangjie, Fuzhou, China/Fujian Key Laboratory of Rehabilitation Technology, 13 Hudong branch Road, Fuzhou, Fujian, China/Fujian Key Laboratoty of Integrative Medicine on Geriatrics, Fujian University of Traditional Chinese Medicine, 1 Qiuyang Road, Minhou Shangjie, Fuzhou, Fujian, China.
Academy of Integrative Medicine, Fujian University of Traditional Chinese Medicine, 1 Qiuyang Road, Minhou Shangjie, Fuzhou, China/Fujian Key Laboratoty of Integrative Medicine on Geriatrics, Fujian University of Traditional Chinese Medicine, 1 Qiuyang Road, Minhou Shangjie, Fuzhou, Fujian, China.
Pak J Pharm Sci. 2020 Jan;33(1(Special)):403-408.
Inflammatory response that occur post-ischemia is a serious problem in the treatment of ischemic brain disease. MicroRNA-155 is a brain-specific or brain-enriched miRNA, which mediates inflammatory reactions in cerebral ischemic tissue by regulating inflammatory signal and the expression level of SOCS1. The present study was aimed to assess the effect of GuaLou GuiZhi Decoction (GLGZD) on miR-155 expression in activated microglia following inflammation and further explore the role of GLGZD on expression of the inflammation-related gene. BV2 cells were used to simulated by LPS to make the inflammatory model. Expression level of miR-155 was detected by Real-Time PCR. BV2 cells after simulated by LPS were then transfected with miR-155 mimic and its negative controls. Cytokines release were measured by corresponding purchased ELISA kits, respectively. Then target protein expression of miR-155 were detected by western blotting assay. After miRNA over expression transfections, expressions of inflammation-related factors, SOCS-1 and SAMD in BV2 cells after activation were measured by Western blot assay. Results showed that in BV2 cells after simulated by LPS, miR-155 was upregulated. The elevated miR-155 expression enhanced the inflammatory cytokine release. miR-155 directly target and negatively regulated SOCS-1 and SMAD-1 expression. Over expression of SOCS-1 and SMAD reduced inflammatory action that was enhanced by miR-155 mimic transfection. miR-155 was positively related with activation of NF-ϰB signal pathways via SOCS-1 and SMAD. In conclusion, GuaLou GuiZhi Decoction (GLGZD) might exert its anti-inflammatory action by inhibiting the expression of miR-155, indicating that miR-155 may be used as a treatment target in clinical treatment with GuaLou GuiZhi Decoction (GLGZD) in ischemic brain.
缺血后发生的炎症反应是缺血性脑疾病治疗中的一个严重问题。微小RNA-155是一种脑特异性或脑富集的微小RNA,它通过调节炎症信号和SOCS1的表达水平来介导脑缺血组织中的炎症反应。本研究旨在评估瓜蒌桂枝汤(GLGZD)对炎症后活化小胶质细胞中miR-155表达的影响,并进一步探讨GLGZD对炎症相关基因表达的作用。采用BV2细胞经脂多糖(LPS)模拟建立炎症模型。通过实时荧光定量PCR检测miR-155的表达水平。经LPS模拟后的BV2细胞再转染miR-155模拟物及其阴性对照。分别用相应购买的酶联免疫吸附测定(ELISA)试剂盒检测细胞因子释放。然后通过蛋白质免疫印迹法检测miR-155的靶蛋白表达。在进行微小RNA过表达转染后,通过蛋白质免疫印迹法检测活化后BV2细胞中炎症相关因子、SOCS-1和SAMD的表达。结果显示,在经LPS模拟后的BV2细胞中,miR-155表达上调。miR-155表达升高增强了炎症细胞因子的释放。miR-155直接靶向并负调控SOCS-1和SMAD-1的表达。SOCS-1和SMAD的过表达减少了miR-155模拟物转染增强的炎症作用。miR-155通过SOCS-1和SMAD与核因子-κB信号通路的激活呈正相关。综上所述,瓜蒌桂枝汤(GLGZD)可能通过抑制miR-155的表达发挥其抗炎作用,表明miR-155可能作为瓜蒌桂枝汤(GLGZD)治疗缺血性脑疾病临床治疗的靶点。