Spinal cord injury(SCI)is one of the most devastating traumas,and the aberrant proliferation of astrocytes usually causes neurological deficits.However,the mechanism underlying astrocyte over-proliferation after SCI i...Spinal cord injury(SCI)is one of the most devastating traumas,and the aberrant proliferation of astrocytes usually causes neurological deficits.However,the mechanism underlying astrocyte over-proliferation after SCI is unclear.Grin2c(glutamate ionotropic receptor type 2c)plays an essential role in cell proliferation.Our bioinformatic analysis indicated that Grin2c and Ca^(2+)transport functions were inhibited in astrocytes after SCI.Suppression of Grin2c stimulated astrocyte proliferation by inhibiting the Ca^(2+)/calmodulin-dependent protein kinase 2b(CaMK2b)pathway in vitro.By screening different inflammatory factors,interleukin 1α(IL1α)was further found to inhibit Grin2c/Ca^(2+)/CaMK2b and enhance astrocyte proliferation in an oxidative damage model.Blockade of IL1αusing neutralizing antibody resulted in increased Grin2c expression and the inhibition of astrocyte proliferation post-SCI.Overall,this study suggests that IL1αpromotes astrocyte proliferation by suppressing the Grin2c/Ca^(2+)/CaMK2b pathway after SCI,revealing a novel pathological mechanism of astrocyte proliferation,and may provide potential targets for SCI repair.展开更多
基金supported by the National Natural Science Foundation of China(82071362,82002899)the Basic Research Project of Shenzhen Science and Technology Innovation Commission(JCYJ202205303001577,JCYJ20190809165201646)Basic Research Projects of Shenzhen Science and Technology Program(JCYJ20180307150610733).
文摘Spinal cord injury(SCI)is one of the most devastating traumas,and the aberrant proliferation of astrocytes usually causes neurological deficits.However,the mechanism underlying astrocyte over-proliferation after SCI is unclear.Grin2c(glutamate ionotropic receptor type 2c)plays an essential role in cell proliferation.Our bioinformatic analysis indicated that Grin2c and Ca^(2+)transport functions were inhibited in astrocytes after SCI.Suppression of Grin2c stimulated astrocyte proliferation by inhibiting the Ca^(2+)/calmodulin-dependent protein kinase 2b(CaMK2b)pathway in vitro.By screening different inflammatory factors,interleukin 1α(IL1α)was further found to inhibit Grin2c/Ca^(2+)/CaMK2b and enhance astrocyte proliferation in an oxidative damage model.Blockade of IL1αusing neutralizing antibody resulted in increased Grin2c expression and the inhibition of astrocyte proliferation post-SCI.Overall,this study suggests that IL1αpromotes astrocyte proliferation by suppressing the Grin2c/Ca^(2+)/CaMK2b pathway after SCI,revealing a novel pathological mechanism of astrocyte proliferation,and may provide potential targets for SCI repair.
文摘目的:制备针对白血病干细胞(LSC)表面标志蛋白IL1RAP(IL-1 receptor accessory protein)的特异性单克隆抗体并对其进行鉴定。方法:杂交瘤细胞3H6E10、10D8A7种植BALB/c小鼠腹腔,收获腹水,纯化后得到特异性抗人IL1RAP的单克隆抗体。分别应用非变性聚丙烯酰胺凝胶电泳(Nondenaturing-PAGE)、ELISA和Western blot法对纯化单克隆抗体分别进行抗体纯度、效价和敏感性检测。结果:获得了两种IL1RAP纯化单克隆抗体,命名为3H6E10 Mc Ab和10D8A7 Mc Ab,其纯度分别为95%和94%;两种纯化的单克隆抗体效价为1∶81000,且能特异性识别IL1RAP纯化蛋白、正常人及白血病病人的细胞内源性蛋白。结论:本研究成功制备了能特异性结合人IL1RAP的纯化单克隆抗体,为将来有效清除体内LSC提供了新途径。
