[1]高皖皎,邓秋狄,白殊同,等.佐剂型关节炎大鼠滑膜成纤维细胞模型建立及特征分析[J].中国药理学通报,2015,(12):1693-1698.[doi:10.3969/j.issn.1001.1978.2015.12.013]
 GAO Wan-jiao,DENG Qiu-di,BAI Shu-tong,et al.Establishment and characteristic analysis of fibroblast-like synoviocytes in rats with adjuvant arthritis[J].Chinese Pharmacological Bulletin,2015,(12):1693-1698.[doi:10.3969/j.issn.1001.1978.2015.12.013]
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佐剂型关节炎大鼠滑膜成纤维细胞模型建立及特征分析()
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《中国药理学通报》[ISSN:/CN:]

卷:
期数:
2015年12期
页码:
1693-1698
栏目:
论 著
出版日期:
2015-12-30

文章信息/Info

Title:
Establishment and characteristic analysis of fibroblast-like synoviocytes in rats with adjuvant arthritis
作者:
高皖皎邓秋狄白殊同佟 丽
南方医科大学中医药学院,广东 广州 510515
Author(s):
GAO Wan-jiao DENG Qiu-di BAI Shu-tong TONG Li
Dept of TMC, Southern Medical University, Guangzhou 510515,China
关键词:
滑膜成纤维细胞 类风湿性关节炎 佐剂型关节炎大鼠 Bcl-2 Bax Cleave-caspase-3
Keywords:
fibroblast-like synoviocytes rheumatoid arthritis adjuvant arthritis rat Bcl-2 Bax Cleave-caspase-3
分类号:
R-332; R322.72; R329.24; R392.12; R593.22; R684.3
DOI:
10.3969/j.issn.1001.1978.2015.12.013
文献标志码:
A
摘要:
目的 建立佐剂型关节炎大鼠滑膜成纤维细胞(AA-FLS)原代培养方法及其特性分析,探讨AA-FLS作为RA体外研究细胞模型的可行性。方法 采用热杀死结核分枝杆菌H37Ra(Mtb)免疫SD 大鼠,建立佐剂型关节炎大鼠模型,取大鼠双侧膝关节滑膜组织进行原代细胞培养,显微镜下观察细胞形态; 免疫细胞化学方法鉴定细胞; CCK-8法检测细胞增殖活性; ELISA方法测定细胞上清中TNF-α和IL-1β水平; Hochest 33258法检测细胞凋亡状态; Western bolt方法检测AA FLS细胞线粒体凋亡通路重要调控蛋白Bcl-2和Bax及Pro-caspase-3和Cleave-caspase-3水平的表达。结果 胶原酶法分离获得的滑膜成纤维细胞经传代纯化,呈梭形的细胞占0.95以上,免疫细胞化学鉴定显示滑膜成纤维细胞vimentin表达呈阳性,CD68表达为阴性; AA模型大鼠滑膜成纤维细胞增殖活性较正常大鼠滑膜成纤维细胞增殖活性明显提高,细胞凋亡率降低; 在培养的细胞上清中TNF-α和IL-1β水平明显升高; 线粒体凋亡通路调控蛋白Bcl-2水平明显升高,而Bax蛋白表达水平明显降低,同时AA-FLS细胞中pro-caspase-3表达量增加。结论 AA-FLS具有增殖活跃、凋亡抑制和分泌高水平促炎性细胞因子的生物特性,可作为体外筛选抗RA药物的细胞模型。
Abstract:
Aims To establish the methods of primary culture of fibroblast-like synoviocytes in rats with adjuvant arthritis(AA-FLS)and analyze the feature and to investigate the possibility of AA-FLS as the model for the RA in vitro. Methods The synovial cellsobtained from the SD rats were immunized by the Mtb and identified by morphology and immunocytochemistry. The viability of AA-FLS was assessed by Cell Counting Kit-8. ELISA was applied to detect TNF-α and IL-1β in cell media. Apoptosis was measured by Hochest 33258.The expressions of mitochondrial apoptosis-related molecules, including Bcl-2, Bax, Pro-caspase-3 and Cleave-caspase-3 were determined by Western Blot. Result In isolated primary synovial cells, more than 95% of AA-FLSs were fusiform. Immunocytochemistry result showed a positive expression of vimentin and a negative expression of CD68 in AA-FLS. Cell proliferation of AA-FLS was higher than FLS and cell apoptosis of AA-FLS was curbed. Western blot data demonstrated that the protein expressions of Bcl-2, Bax were regulated and the expression of caspase-3 was activated in AA-FLS. Conclusions AA-FLS is biologically characterized by high level proliferation activity and inflammatory cytokines and apoptosis suppression. AA-FLS can be used as the model for the RA in vitro.

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更新日期/Last Update: 1900-01-01