中国组织工程研究2017，Vol.21Issue(4)：527-531,5.DOI:10.3969/j.issn.2095-4344.2017.04.006
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
Establishment and identification of the rat models of chondrocyte apoptosis induced by tumor necrosis factor-alpha
摘要
Abstract
BACKGROUND:Tumor necrosis factor-α(TNF-α), a main cytokine inducing chondrocyte apoptosis of osteoarthritis, plays a regulatory role in the process of osteoarthritis. OBJECTIVE:To compare the rat models of chondrocyte apoptosis induced by different mass concentrations of TNF-α, thus providing theoretical basis for further study on the regulation of drugs on chondrocyte apoptosis. METHODS:Chondrocytes were isolated from the knee cartilage of 4-week-old Sprague-Dawley rats of clean grade by mechanical l col agenase digestion and were then incubated with different mass concentrations of TNF-αto induce apoptosis. The morphology of chondrocytes was observed under inverted phase contrast microscope, cel s were identified using immunohistochemical staining of type II col agen, as wel as the cel viability and apoptosis were detected by MTT and DAPI, respectively. RESULTS AND CONCLUSION:(1) In vitro, the cytoplasm of chondrocytes was stained brown-yel ow by using immunohistochemical staining of type II col agen. (2) At 48 hours, the apoptosis rate of chondrocytes induced by 10, 20 and 30μg/L TNF-αwas significantly higher than that of the 0μg/L TNF-α(P<0.01), and the apoptosis rate of chondrocytes induced by 40μg/L TNF-αwas significantly higher than that of the 10μg/L TNF-α(P<0.01). (3) The viability of chondrocytes induced by 10, 20 and 40μg/L TNF-αwas significantly lower than that of the 0μg/L TNF-α(P<0.01). In detail, the viability of chondrocytes induced by 20μg/L TNF-αwas lower than that of the 10μg/L TNF-α(P<0.05);the viability of chondrocytes induced by 40μg/L TNF-αwas significantly lower than that of the 10 and 20μg/L TNF-α(P<0.01, P<0.05). (4) These results suggest that 20μg/L TNF-αcan successful y induce the chondrocyte apoptosis model.关键词
组织构建/软骨细胞/肿瘤坏死因子α/MTT/DAPI/Ⅱ型胶原鉴定/凋亡模型/炎症/国家自然科学基金分类
医药卫生引用本文复制引用
陈后煌,邵翔,李俐,吴明霞,李西海..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[J].中国组织工程研究,2017,21(4):527-531,5.基金项目
国家自然科学基金(81373719)；陈可冀中西医结合发展基金(CKJ2015009)；福建省科技厅科技计划项目(2015J01479) （81373719）
