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首页> 外文期刊>American Journal of Physiology >A 78-kDa glucose-regulated protein is involved in the decrease of interleukin-6 secretion by lead treatment from astrocytes.
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A 78-kDa glucose-regulated protein is involved in the decrease of interleukin-6 secretion by lead treatment from astrocytes.

机译：通过星形胶质细胞的先导处理，一个78 kDa的葡萄糖调节蛋白参与了白细胞介素6分泌的减少。

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摘要

Interleukin (IL)-6 is a cytokine produced mainly by microglia and astrocytes and plays a pleiotropic role in the central nervous system. In this study, we cloned rat IL-6 cDNA into an enhanced green fluorescent protein (EGFP) or a red fluorescent protein (DsRed2) vector and rat 78-kDa glucose-regulated protein (GRP78) cDNA into an EGFP vector to construct IL-6-EGFP, IL-6-DsRed2, and GRP78-EGFP chimeras for the investigation of the mechanism of IL-6 secretion from astrocytes. The data showed that constructed IL-6-EGFP and IL-6-DsRed2 chimeras retained the secretory property, and the secretion of IL-6-EGFP from astrocytes could be attenuated by GRP78 depletion with double-stranded RNA interference. Coexpression of IL-6-DsRed2 and dysfunctional GRP78-EGFP abolished IL-6-DsRed2 secretion, and two chimeric proteins colocalized inside living astrocytes. Coimmunoprecipitation analysis indicated that IL-6 and GRP78 resided in the same complex. The data further revealed that IL-6-EGFP secretion from astrocytes was blocked by the heavy metal lead (Pb) in a concentration-dependent manner. Analysis of the Pb interaction with protein on a Pb-affinity column demonstrated that Pb bound to GRP78 but failed to bind to IL-6. Therefore, these data suggest that IL-6-EGFP or IL-6-DsRed2 chimeras can be used as imaging probes to study IL-6 secretion from living cells, that GRP78 is involved in IL-6 secretion from astrocytes, and that Pb can block IL-6 secretion from astrocytes via targeting GRP78.
机译：白介素（IL）-6是主要由小胶质细胞和星形胶质细胞产生的细胞因子，在中枢神经系统中具有多效作用。在这项研究中，我们将大鼠IL-6 cDNA克隆到增强的绿色荧光蛋白（EGFP）或红色荧光蛋白（DsRed2）载体中，并将​​大鼠78-kDa葡萄糖调节蛋白（GRP78）cDNA克隆到EGFP载体中以构建IL- 6-EGFP，IL-6-DsRed2和GRP78-EGFP嵌合体用于研究星形胶质细胞分泌IL-6的机制。数据显示，构建的IL-6-EGFP和IL-6-DsRed2嵌合体保留了分泌特性，而GRP78消耗和双链RNA干扰可减弱星形胶质细胞中IL-6-EGFP的分泌。 IL-6-DsRed2和功能异常的GRP78-EGFP的共表达消除了IL-6-DsRed2的分泌，并且两个嵌合蛋白共定位在活的星形胶质细胞内部。免疫共沉淀分析表明IL-6和GRP78驻留在同一复合物中。数据进一步揭示，星形胶质细胞分泌的IL-6-EGFP被重金属铅（Pb）浓度依赖性地阻断。在铅亲和柱上分析铅与蛋白质的相互作用，结果表明铅与GRP78结合，但与IL-6结合失败。因此，这些数据表明IL-6-EGFP或IL-6-DsRed2嵌合体可以用作成像探针，以研究活细胞的IL-6分泌，GRP78参与星形胶质细胞的IL-6分泌，而铅可以通过靶向GRP78阻止星形胶质细胞分泌IL-6。

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