Raw data for "Comparison of the effects between catalase and superoxide dismutase on regulating macrophage inflammatory response and protecting osteogenic function of periodontal ligament cells"

Background: Reactive oxygen species (ROS) have been confirmed closely associated with the pathological process of periodontitis, but the specific roles played by different ROS types are still to be investigated. Catalase (CAT) and Superoxide dismutase (SOD) specifically eliminate hydrogen peroxide (H2O2) and superoxide anion (O2•-), respectively. We for the first time compare the effects and mechanisms of CAT and SOD in protecting periodontal ligament cells (PDLCs) against oxidative damage, reducing the expression of macrophage inflammatory factors, and preserving the osteogenic differentiation function of PDLCs by modulating the inflammatory environment.Methods: CAT or SOD in combination with lipopolysaccharide (LPS) were added to the culture medium of RAW 264.7 and PDLCs. The intracellular ROS level, lipid peroxidation and DNA damage were observed by confocal microscope. Inflammation levels were assessed by real-time quantitative polymerase chain reaction (RT-qPCR) and western blot. A co-culture system of macrophages and PDLCs was established, and the osteogenic differentiation of PDLCs was evaluated by alkaline phosphatase staining, alizarin red S staining, RT-qPCR and Western blot. Finally, differentially expressed genes (DEGs) in CAT and SOD were detected by RNA sequencing and the biological functions and signaling pathways involved were analyzed.Results: CAT or SOD can effectively inhibit intracellular ROS levels, lipid peroxidation and DNA damage, as well as increase the levels of antioxidative molecules and decrease the levels of inflammatory factors. SOD increased the levels of antioxidative molecules more strongly, while CAT reduced inflammatory factors more effectively. The RNA sequencing results indicate that CAT exhibits stronger inhibitory effects on inflammation-related signaling pathways, which could account for the above-mentioned differences.Conclusions: In this study, we observed differential antioxidant and anti-inflammatory effects between CAT and SOD, which may be associated with CAT's better inhibition of the activation of inflammatory pathways. Our study will provide scientific references for the future development of highly selective ROS- scavenging antioxidant drugs.

研究背景：活性氧（Reactive oxygen species, ROS）已被证实与牙周炎的病理进程密切相关，但不同类型ROS所发挥的具体作用仍有待研究。过氧化氢酶（Catalase, CAT）与超氧化物歧化酶（Superoxide dismutase, SOD）可分别特异性清除过氧化氢（Hydrogen peroxide, H₂O₂）与超氧阴离子（Superoxide anion, O₂•⁻）。本研究首次对比了CAT与SOD在保护牙周膜细胞（Periodontal ligament cells, PDLCs）免受氧化损伤、降低巨噬细胞炎症因子表达，以及通过调控炎症环境维持PDLCs成骨分化功能方面的作用与机制。
研究方法：将CAT或SOD与脂多糖（Lipopolysaccharide, LPS）共同添加至RAW 264.7细胞与PDLCs的培养基中。通过共聚焦显微镜观察细胞内ROS水平、脂质过氧化与DNA损伤情况；采用实时定量聚合酶链反应（Real-time quantitative polymerase chain reaction, RT-qPCR）与蛋白质印迹（Western blot）评估炎症水平。构建巨噬细胞与PDLCs的共培养体系，并通过碱性磷酸酶染色、茜素红S染色、RT-qPCR及蛋白质印迹对PDLCs的成骨分化能力进行评价。最终通过RNA测序检测CAT与SOD处理组的差异表达基因（Differentially expressed genes, DEGs），并分析其涉及的生物学功能与信号通路。
研究结果：CAT与SOD均可有效抑制细胞内ROS水平、脂质过氧化与DNA损伤，同时提升抗氧化分子水平并降低炎症因子表达。其中SOD对抗氧化分子水平的提升效果更为显著，而CAT对炎症因子的抑制作用更强。RNA测序结果显示，CAT对炎症相关信号通路的抑制作用更强，这或可解释上述差异。
研究结论：本研究发现CAT与SOD在抗氧化与抗炎作用上存在差异，这可能与CAT更高效地抑制炎症通路激活有关。本研究可为未来开发高选择性ROS清除类抗氧化药物提供科学参考。