橙皮素抑制氧化应激影响软骨细胞的炎性退变  

Effect of hesperetin on inflammatory degeneration of chondrocytes by inhibiting oxidative stress

作　　者：罗善超 唐继仁[2] Luo Shanchao;Tang Jiren(Guangxi University of Chinese Medicine,Nanning 530011,Guangxi Zhuang Autonomous Region,China;Yulin Orthopedic Hospital of Integrated Traditional Chinese and Western Medicine,Yulin 537000,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西中医药大学,广西壮族自治区南宁市530011 [2]玉林市中西医结合骨科医院,广西壮族自治区玉林市537000

出　　处：《中国组织工程研究》2024年第26期4184-4188,共5页Chinese Journal of Tissue Engineering Research

基　　金：广西自然科学基金资助项目(2019GXNSFAA185060),项目参与人:罗善超。

摘　　要：背景:研究表明,橙皮素可以通过多种机制或者信号通路对软骨细胞产生保护作用,但其保护机制尚未完全阐明。目的:探讨橙皮素对脂多糖诱导的软骨细胞炎性退变的影响。方法:体外提取、培养SD乳鼠关节软骨细胞,采用番红O染色鉴定。通过MTT法检测细胞毒性确定最佳橙皮素干预浓度。将软骨细胞随机分为3组:对照组、模型组和实验组,后2组采用脂多糖诱导软骨细胞建立骨关节炎细胞模型,实验组造模后给予橙皮素干预24 h。采用Calcein-AM/EthD-Ⅰ染色检测细胞活性,免疫组化染色检测软骨特异性Ⅱ型胶原的表达,活性氧检测试剂盒检测软骨细胞内活性氧水平,ELISA检测软骨细胞内抗氧化剂总谷胱甘肽水平,qRT-PCR检测炎症基因白细胞介素1β、白细胞介素6、肿瘤坏死因子α和软骨特异性基因Ⅱ型胶原基因的表达。结果与结论:①番红O染色结果表明,提取的细胞为软骨细胞;②细胞毒性实验表明0.5μmol/L橙皮素干预软骨细胞活力最明显;③与对照组比较,模型组软骨细胞增殖能力降低,活性氧水平升高,总谷胱甘肽水平下降,Ⅱ型胶原降解增加,白细胞介素1β、白细胞介素6和肿瘤坏死因子α基因表达水平升高,软骨特异性Ⅱ型胶原基因表达水平下降,差异均有显著性意义(P<0.05);④与模型组比较,实验组软骨细胞增殖能力上升,活性氧水平下降,总谷胱甘肽水平上升,Ⅱ型胶原降解减少,白细胞介素1β、白细胞介素6和肿瘤坏死因子α基因表达水平下降,软骨特异性Ⅱ型胶原基因表达水平升高(P<0.05)。结果表明:橙皮素对脂多糖诱导的骨关节炎软骨细胞炎性退变具有保护作用,其机制可能与橙皮素抑制活性氧介导的氧化应激有关。BACKGROUND:Studies have shown that hesperetin exerts protective effects on chondrocytes through a variety of mechanisms or signaling pathways,but the protective mechanisms have not been fully elucidated.OBJECTIVE:To investigate the effect of hesperetin on lipopolysaccharide-induced inflammatory degeneration of chondrocytes.METHODS:Knee joint chondrocytes from suckling Sprague-Dawley rats were isolated and cultured in vitro,and identified by Safranine O staining.The cytotoxicity of hesperetin on chondrocytes was determined by the MTT assay to ensure the optimal concentration of hesperetin.Chondrocytes were randomly divided into three groups,including the control group,model group,and experimental group.The cellular model of osteoarthritis was established in the latter two groups by simulating chondrocytes with lipopolysaccharide.Chondrocytes in the experimental group was then intervened with hesperetin for 24 hours.Calcein-AM/EthD-I staining was used to detect cell viability.Immunohistochemical staining was performed to determine the expression of type Ⅱ collagen in chondrocytes.Intracellular reactive oxygen species level was detected by a reactive oxygen species detection kit.Total glutathione level was detected by ELISA.Real-time fluorescent PCR was employed to detect the expression of interleukin 1β,interleukin 6,tumor necrosis factorαand type Ⅱ collagen.RESULTS AND CONCLUSION:Safranine O staining results showed that the cells extracted were chondrocytes.Cytotoxicity test results showed 0.5μmol/L hesperetin had the most significant effect on chondrocyte vitality.Compared with the control group,the model group showed a decrease in chondrocyte proliferation ability,an increase in reactive oxygen species levels,a decrease in total glutathione levels,an increase in type Ⅱ collagen degradation,an increase in the levels of interleukin 1β,interleukin 6,and tumor necrosis factorα,and a decrease in the expression of type Ⅱ collagen(P<0.05).Compared with the model group,in the experimental group,the prolifer

关 键 词：软骨细胞 橙皮素 活性氧 氧化应激 炎症 软骨退变 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R684.3