| 益气化瘀方减轻HIF-1α条件性基因敲除小鼠膝关节软骨退变的研究 |
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投稿时间：2012-11-16
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| 作者 | Author | 单位 | Address | E-Mail |
| 王晶 |
WANG Jing |
上海中医药大学脊柱病研究所 上海中医药大学附属龙华医院, 上海 200032 |
Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China |
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| 董芳芳 |
DONG Fang-fang |
上海中医药大学脊柱病研究所 上海中医药大学附属龙华医院, 上海 200032 |
Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China |
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| 李晓锋 |
LI Xiao-feng |
上海中医药大学脊柱病研究所 上海中医药大学附属龙华医院, 上海 200032 |
Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China |
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| 舒冰 |
SHU Bing |
上海中医药大学脊柱病研究所 上海中医药大学附属龙华医院, 上海 200032 |
Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China |
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| 施杞 |
SHI Qi |
上海中医药大学脊柱病研究所 上海中医药大学附属龙华医院, 上海 200032 |
Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China |
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| 王拥军 |
WANG Yong-Jun |
上海中医药大学脊柱病研究所 上海中医药大学附属龙华医院, 上海 200032 |
Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China |
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| 周重建 |
ZHOU Chong-jian |
上海中医药大学脊柱病研究所 上海中医药大学附属龙华医院, 上海 200032 |
Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China |
zhouchongjian@hotmail.com |
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| 期刊信息：《中国骨伤》2013年，第26卷，第6期，第486-492页 |
| DOI：10.3969/j.issn.1003-0034.2013.06.011 |
| 基金项目:国家自然基金面上项目(编号:30973760);上海市高校创新团队计划(编号:沪教委科[2009]6号);国家十二五“重大新药创制”专项项目(编号:2011ZX09302-006-04) |
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中文摘要:
目的: 观察低氧诱导因子-1α(HIF-1α)条件性基因敲除(HIF-1α cKO)小鼠膝关节软骨退变情况,以及中药复方益气化瘀方对减缓HIF-1α cKO小鼠椎间盘软骨退变的作用。
方法: ①将杂交繁殖获得同窝的HIF-1α +/+小鼠和HIF-1α-/-小鼠,分为HIF-1α +/+小鼠4月龄组、HIF-1α-/-小鼠4月龄组,HIF-1α +/+小鼠6月龄组、HIF-1α-/-小鼠6月龄组,每组3只。在4、6月龄依次处死相应的小鼠,取膝关节软骨,分别进行Mankin评分、藏红固绿、HE及免疫组化相关指标染色及分析,研究软骨组织的改变情况。②将2月龄HIF-1α-/-小鼠随机分为生理盐水组和益气化瘀方组,以生理盐水组小鼠作为正常对照,每组6只,共12只。灌胃给药2个月后,取各组小鼠的膝关节,分别进行Mankin评分、HE染色和藏红固绿染色,Col Ⅱ、Col X、VEGF、MMP-13和Sox9免疫组化染色及分析。
结果: ①HIF-1α-/-小鼠4月龄组膝关节软骨组织出现破损和骨化,细胞分布不均匀,软骨细胞减少。HIF-1α-/-小鼠6月龄组膝关节软骨损伤更加严重。HIF-1α-/-小鼠4月龄组椎间盘软骨中Ⅱ型胶原(Col Ⅱ)和Sox9表达降低,X型胶原(Col X)、基质金属蛋白酶-13(MMP-13)、VEGF蛋白表达增加。HIF-1α-/-小鼠6月龄组Col Ⅱ蛋白与Sox9表达进一步降低,Col X、MMP13、VEGF蛋白表达进一步上升。②与生理盐水组比较,益气化瘀方干预后膝关节软骨部位的骨化和缺损减轻,软骨细胞分布较均匀,细胞总数增加。免疫组化染色显示,益气化瘀方组Col Ⅱ和Sox9蛋白的表达升高,Col X、VEGF、MMP-13蛋白表达减少。
结论: HIF-1α基因敲除小鼠出现了膝关节软骨退变,并且这种退变随着小鼠增龄而加重;益气化瘀方可以减轻HIF-1α基因敲除小鼠的膝关节软骨退变。 |
| 【关键词】HIF-1α基因 小鼠 膝关节 软骨 中草药 |
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| Effects of Yiqi Huayu prescription(益气化瘀方) on knee cartilage degeneration in HIF-1α gene knockout mice |
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ABSTRACT
Objective: To study the role of hypoxia-inducible factor 1 alpha (HIF1α) on knee cartilage degeneration,and to explore the effects and mechanisms of Chinese herbal compound Yiqi Huayu prescription(益气化瘀方) on HIF-1αgene knockout mice on knee cartilage degeneration.
Methods: The 4-month and 6-month HIF-1α gene knock out mice were obtained by interbreeding,and divided into HIF-1α +/+ 4-month mice group,HIF-1α-/-4-month mice group,HIF-1α +/+ 6-month mice group and HIF-1α-/-6-month mice group,3 mice in each group. And then the 2-month-old HIF-1α gene knock out mice were randomly divided into Yiqi Huayu prescription(益气化瘀方) group and physiological saline group. There were 6 mice in each group. After 2 months' drug administration,the knee joint of mice was collected,and the Mankin score were evaluated;Safranine-fast green staining,HE Staining,and immunohistochemistry analysis for VEGF,Col X,ColⅡ,MMP-13 and Sox-9 were performed erespectively.
Results: ①Compared to the results in the HIF-1α+/+ mice groups,the HIF-1α-/-mice developed aging related cartilage loss and bony tissue appearance,cartilage defects increased,and cells reduced. In HIF-1α-/-4-month mice and 6-month mice group,the expresion of Col Ⅱand Sox9 decreased,and the expression of Col X,MMP-13 and VEGF increased. ②Compared to the physiological saline group,the ossification and defect of knee joint cartilage reduced of mice in the Yiqi Huayu prescription(益气化瘀方) group,the cartilage cell distribution was more uniform,and the total number of cells increased. The expression of type Ⅱ collagen and Sox9 protein increased,expression of Col X,MMP-13 and VEGF protein decreased of mice in the Yiqi Huayu prescription(益气化瘀方) group.
Conclusion: The knee cartilage degenerates in the HIF-1α cKO mice,and the degeneration increased with age adding. The Yiqi Huayu prescription(益气化瘀方) can delay the degeneration of knee cartilage of HIF-1α cKO mice. |
| KEY WORDS HIF-1α gene Mice Knee joint Cartilage Drugs,Chinese herbal |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 王晶,董芳芳,李晓锋,舒冰,施杞,王拥军,周重建.益气化瘀方减轻HIF-1α条件性基因敲除小鼠膝关节软骨退变的研究[J].中国骨伤,2013,26(6):486~492 |
| 英文格式: | WANG Jing,DONG Fang-fang,LI Xiao-feng,SHU Bing,SHI Qi,WANG Yong-Jun,ZHOU Chong-jian.Effects of Yiqi Huayu prescription(益气化瘀方) on knee cartilage degeneration in HIF-1α gene knockout mice[J].zhongguo gu shang / China J Orthop Trauma ,2013,26(6):486~492 |
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