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首页> 外文期刊>Cellular Physiology and Biochemistry >Advanced Oxidation Protein Products Inhibit Proliferation and Differentiation of Rat Osteoblast-like Cells via NF-kappa B Pathway
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Advanced Oxidation Protein Products Inhibit Proliferation and Differentiation of Rat Osteoblast-like Cells via NF-kappa B Pathway

机译：先进的氧化蛋白产物通过NF-κB途径抑制大鼠成骨样细胞的增殖和分化

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Background/Aims: Advanced oxidation protein products (AOPPs), a novel marker for oxidative protein damage, regulated cell behavior in many cell types. Although previous reports have revealed that oxidative stress resulted in an increased AOPPs content in cultured osteoblast-like cells, the effects of AOPPs on osteoblast-like cell function remain unclear. This study aimed to investigate the effect of AOPPs on the proliferation and differentiation of rat osteoblast-like ( ROB) cells in vitro. Methods: ROB cells isolated from calvarias of newborn rats were incubated with AOPPs-modified rat serum albumin (AOPPs-RSA) prepared in vitro by incubation of RSA with hypochlorous acid. Cell proliferation was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyl tetrazolium bromide (MTT) assay. Alkaline phosphatase activity was measured. Expression of osteocalcin mRNA and protein were examined using reverse transcriptase-polymerase chain reaction and Western blot, respectively. Reactive oxygen species (ROS) generation was detected by flow cytometry and fluorescent microscope. Phosphorylation of nuclear factor (NF)-kappa B p65 subunit was detected by Western blot. Results: Exposure of ROB cells to AOPPs-RSA significantly inhibited cell proliferation, decreased alkaline phosphatase activity, down-regulated the expression of osteocalcin mRNA and protein in both a concentration- and time-dependent manner. AOPPs challenge induced ROS generation and NF-kappa B p65 phosphorylation, which were inhibited by superoxide dismutase, catalase and NADPH oxidase inhibitors diphenyleneiodonium and apocynin. Furthermore, NF-kappa B inhibitor SN50 could reverse the AOPPs-induced inhibition of ROB cell proliferation and differentiation. Conclusion: These results suggest that AOPPs can inhibit proliferation and differentiation of ROB cells through ROS-dependent NF-kappa B pathway. Copyright
机译：背景/目的：先进的氧化蛋白产品（AOPPs），一种氧化蛋白损伤的新标记，可调节多种细胞类型的细胞行为。尽管以前的报道表明氧化应激导致培养的成骨细胞样细胞中AOPPs含量增加，但是AOPPs对成骨细胞样细胞功能的影响仍不清楚。这项研究旨在调查AOPPs在体外对大鼠成骨样（ROB）细胞增殖和分化的影响。方法：将从新生大鼠颅盖分离出的ROB细胞与AOPPs修饰的大鼠血清白蛋白（AOPPs-RSA）一起孵育，该蛋白是通过将RSA与次氯酸孵育而体外制备的。通过3-（4，5-二甲基噻唑-2-基）-2，5-二苯基溴化四唑（MTT）测定法评估细胞增殖。测量碱性磷酸酶活性。分别使用逆转录酶-聚合酶链反应和蛋白质印迹法检测骨钙素mRNA和蛋白的表达。通过流式细胞仪和荧光显微镜检测活性氧（ROS）的产生。通过蛋白质印迹检测核因子（NF）-κBp65亚基的磷酸化。结果：ROB细胞暴露于AOPPs-RSA显着抑制细胞增殖，降低碱性磷酸酶活性，并以浓度和时间依赖性方式下调骨钙素mRNA和蛋白的表达。 AOPP挑战诱导的ROS生成和NF-κB p65磷酸化，这被超氧化物歧化酶，过氧化氢酶和NADPH氧化酶抑制剂二苯并碘鎓和载脂蛋白抑制。此外，NF-κB抑制剂SN50可以逆转AOPPs对ROB细胞增殖和分化的抑制作用。结论：这些结果表明，AOPPs可以通过ROS依赖性的NF-κB途径抑制ROB细胞的增殖和分化。版权

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