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Apelin 通过 PI-3K/Akt 和 MEK/Erk 通路激活小胶质细胞 BV2 细胞中炎症细胞因子的表达。

Apelin activates the expression of inflammatory cytokines in microglial BV2 cells via PI-3K/Akt and MEK/Erk pathways.

机构信息

Department of Ophthalmology, People' Hospital, Peking University, Beijing, 100044, China.

出版信息

Sci China Life Sci. 2015 Jun;58(6):531-40. doi: 10.1007/s11427-015-4861-0. Epub 2015 May 8.

Abstract

This paper aims to observe the changes of the inflammatory cytokines in microglial BV2 cells stimulated by apelin, and investigate the mechanism of inflammatory cytokines secretion after apelin stimulation. Immunofluorescence and quantitative real-time PCR were performed to observe expression of TNF-α, IL-1β, IL-10, MIP-1α, and MCP-1 in BV2 cells. Western blot was used to investigate the expression of phosphorylation PI-3K/Akt and phosphorylation Erk signaling pathways in BV2 cells after stimulation by apelin. Furthermore, PI-3K/Akt inhibitor (LY294402) and Erk inhibitor (U0126) were used as antagonists to detect the secretion mechanisms of cytokines in BV2 cells stimulated by apelin. Exogenous recombinant apelin activated the expression of TNF-α, IL-1β, MCP-1 and MIP-1α in BV2 cells by the detection of fluorescence expression and mRNA. Apelin also unregulated the protein expression of p-PI-3K/Akt and p-Erk in BV2 cells induced by apelin. LY294002 and U0126 inhibited activation of p-PI-3K/Akt and p-Erk expression by Western blot and attenuated the expression of inflammation factors in BV2 cells by fluorescence staining. This study demonstrates that apelin is a potential activator of inflammation factors through the PI3K/Akt and Erk signaling pathway and is potential therapeutically relevant to inflammatory responses of microglia cells.

摘要

本文旨在观察促胃液素原(apelin)刺激小胶质细胞 BV2 后炎症细胞因子的变化,探讨促胃液素原刺激后炎症细胞因子分泌的机制。采用免疫荧光和实时定量 PCR 观察 TNF-α、IL-1β、IL-10、MIP-1α 和 MCP-1 在 BV2 细胞中的表达。采用 Western blot 检测促胃液素原刺激后 BV2 细胞中磷酸化 PI-3K/Akt 和磷酸化 Erk 信号通路的表达。进一步采用 PI-3K/Akt 抑制剂(LY294002)和 Erk 抑制剂(U0126)作为拮抗剂,检测促胃液素原刺激的 BV2 细胞中细胞因子的分泌机制。外源性重组促胃液素原通过荧光表达和 mRNA 检测激活 BV2 细胞中 TNF-α、IL-1β、MCP-1 和 MIP-1α 的表达。促胃液素原还可调节 BV2 细胞中促胃液素原诱导的 p-PI-3K/Akt 和 p-Erk 蛋白表达。LY294002 和 U0126 通过 Western blot 抑制 p-PI-3K/Akt 和 p-Erk 表达的激活,并通过荧光染色减弱 BV2 细胞中炎症因子的表达。本研究表明,促胃液素原通过 PI3K/Akt 和 Erk 信号通路是炎症因子的潜在激活剂,对小胶质细胞的炎症反应具有潜在的治疗相关性。

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