肿瘤坏死因子α诱导大鼠软骨细胞凋亡模型的建立及鉴定

被引:12
作者
陈后煌 [1 ]
邵翔 [1 ]
李俐 [2 ]
吴明霞 [2 ]
李西海 [1 ]
机构
[1] 福建中医药大学中西医结合研究院
[2] 福建中医药大学附属第二人民医院
关键词
软骨细胞; 肿瘤坏死因子α; 凋亡模型; 炎症; 组织工程; 组织构建; MTT; DAPI; Ⅱ型胶原鉴定; 国家自然科学基金;
D O I
暂无
中图分类号
R684 [关节疾病及损伤];
学科分类号
100220 [骨科学];
摘要
背景:肿瘤坏死因子α是诱导骨关节炎软骨细胞凋亡的主要细胞因子,对骨关节炎的病理进程具有重要调节作用。目的:比较不同质量浓度肿瘤坏死因子α诱导大鼠软骨细胞凋亡模型,为进一步探讨药物对软骨细胞凋亡的调控作用提供理论支持。方法:清洁级4周龄SD大鼠,采用机械-Ⅱ型胶原酶消化法获取膝关节软骨细胞,并用不同质量浓度的肿瘤坏死因子α(0,10,20,30,40μg/L)诱导建立软骨细胞的凋亡模型,倒置相差显微镜观察软骨细胞的形态结构,Ⅱ型胶原免疫组化鉴定软骨细胞,MTT检测软骨细胞的活性,DAPI检测软骨细胞的凋亡情况。结果与结论:(1)体外培养软骨细胞,Ⅱ型胶原免疫组化染色可见胞浆呈棕黄色,为阳性细胞;(2)软骨细胞凋亡率,干预48 h肿瘤坏死因子α10,20,30μg/L明显高于0μg/L(P<0.01),肿瘤坏死因子α40μg/L明显高于10μg/L(P<0.01);(3)软骨细胞活性,干预48 h肿瘤坏死因子α10,20,40μg/L明显低于0μg/L(P<0.01),肿瘤坏死因子α20μg/L明显低于10μg/L(P<0.05),肿瘤坏死因子α40μg/L明显低于10μg/L(P<0.01)、20μg/L(P<0.05)。(4)结果说明,20μg/L肿瘤坏死因子α能成功建立炎症介导软骨细胞凋亡模型。
引用
收藏
页码:527 / 531
页数:5
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