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-1αcKO）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-1αcKO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,ColⅡ、ColX、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（ColⅡ）和Sox9表达降低,X型胶原（ColX）、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ColⅡ蛋白与Sox9表达进一步降低,ColX、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ColⅡ和Sox9蛋白的表达升高,ColX、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。

英文摘要：

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 ob- tained 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,Col Ⅱ ,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 delaythe degeneration of knee cartilage of HIF-1?