| 体外IL-β诱导股骨头建立软骨退变模型 |
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Received:October 13, 2014
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| 作者 | Author | 单位 | Unit | E-Mail |
| 丁道芳 |
DING Dao-fang |
上海中医药大学附属曙光医院石氏伤科医学中心, 上海 201203
上海市中医药研究院骨伤科研究所, 上海 201203 |
Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai 201203, China |
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| 庞坚 |
PANG Jian |
上海中医药大学附属曙光医院石氏伤科医学中心, 上海 201203
上海市中医药研究院骨伤科研究所, 上海 201203 |
Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai 201203, China |
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| 宋奕 |
SONG Yi |
苏州市中西医结合医院, 江苏 苏州 215101 |
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| 杜国庆 |
DU Guo-qing |
上海中医药大学附属曙光医院石氏伤科医学中心, 上海 201203
上海市中医药研究院骨伤科研究所, 上海 201203 |
Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai 201203, China |
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| 曹月龙 |
CAO Yue-long |
上海中医药大学附属曙光医院石氏伤科医学中心, 上海 201203
上海市中医药研究院骨伤科研究所, 上海 201203 |
Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai 201203, China |
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| 詹红生 |
ZHAN Hong-sheng |
上海中医药大学附属曙光医院石氏伤科医学中心, 上海 201203
上海市中医药研究院骨伤科研究所, 上海 201203 |
Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai 201203, China |
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| 郑昱新 |
ZHENG Yu-xin |
上海中医药大学附属曙光医院石氏伤科医学中心, 上海 201203
上海市中医药研究院骨伤科研究所, 上海 201203 |
Shi's Center of Orthopedics and Traumatology, Shuguang Hospital Affiliated to Shanghai University of TCM, Shanghai 201203, China |
sg_zyx1728@126.com |
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| 期刊信息：《中国骨伤》2015年28卷，第7期，第648-653页 |
| DOI：10.3969/j.issn.1003-0034.2015.07.015 |
| 基金项目:国家自然科学基金 (No. 81073114,81072830,81373665);上海市教委创新项目(No. 11YZ64);上海市高校青年教师培养资助计划(No. ZZszy12017) |
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目的:IL-1β诱导体外培养大鼠的股骨头致其快速退变,为药物筛选和治疗提供可靠的软骨退变模型。
方法:取2周龄SD幼鼠的股骨头,左右股骨头对应为对照组和实验组,分别培养于正常培养基和诱导培养基中(其中含50 ng/ml的IL-1β),每2 d换液1次,至第3天取出股骨头,固定于4%多聚甲醛,经过脱钙和脱水、石蜡包埋、切片后经甲苯胺蓝和臧红-固绿染色及细胞免疫荧光检测软骨标志基因Ⅱ型胶原和Sox9以及软骨退变相关基因MMP13和ADAMTS5等基因的原位表达。
结果:实验组的股骨头外缘,在培养3 d后甲苯胺蓝染色明显淡染,臧红-固绿染色阳性,提示软骨退变。免疫荧光显示Ⅱ型胶原和Sox9表达减弱,MMP13和ADAMTS5表达增强。
结论:IL-1β培养大鼠股骨头3 d 即诱导其退变,建立软骨组织体外培养诱导其退变的模型。 |
| [关键词]:股骨头 染色 蛋白表达 软骨退变 |
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| Etablishment of cartilage degeneration model by IL-1β in vitro |
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Abstract:
Objective: To establish a reliable model for drug screening and therapy by culturing rat femoral head and inducing cartilage degeneration quickly in vitro.
Methods: The femoral heads from the same SD rats of two-month old were divided into control group and experimental group respectively. They were cultured with DMEM medium plus 10% fetal bovine serum or DMEM medium plus 10% fetal bovine serum plus 50 ng/ml IL-1β for three days. Femoral heads were fixed in 4% paraformaldehyde,decalcified,dehydrated,embedded in paraffin and cut into slices. Specimens were stained with Toluidine blue and Safranine O-Fast Green FCF. The protein expression levels of type Ⅱ collagen,MMP13,Sox9 and ADAMTS5 were analyzed by immunofluorescence.
Results: Both the Toluidine blue and Safranine O staining were pale in the margin of femoral heads which were stimulated with IL-1β for three days compared to that in control group. The Fast Green FCF staining was positive at the edge of the femoral head in experimental group,which indicated that cartilage became degenerated. The expression levels of both type Ⅱ collagen and Sox9 were decreased significantly while the expression levels of MMP13 and ADAMTS5 were increased in experimental group.
Conclusion: The model of cartilage degeneration is established by culturing and inducing the degeneration of the femoral heads quickly in vitro. |
| KEYWORDS:Femoral head Staining Protein expression Cartilage degeneration |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 丁道芳,庞坚,宋奕,杜国庆,曹月龙,詹红生,郑昱新.体外IL-β诱导股骨头建立软骨退变模型[J].中国骨伤,2015,28(7):648~653 |
| 英文格式: | DING Dao-fang,PANG Jian,SONG Yi,DU Guo-qing,CAO Yue-long,ZHAN Hong-sheng,ZHENG Yu-xin.Etablishment of cartilage degeneration model by IL-1β in vitro[J].zhongguo gu shang / China J Orthop Trauma ,2015,28(7):648~653 |
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