益气化瘀方对膝骨关节炎大鼠关节软骨退变的防治作用

目的：研究益气化瘀方对膝骨关节炎大鼠模型关节软骨退变的防治作用及其机制。 方法：1月龄SD大鼠90只，随机分为正常对照组、模型组和益气化瘀方组，每组各30只。模型组，益气化瘀方组大鼠通过肩关节离断术＋直立位诱导法建立大鼠膝骨关节炎模型。益气化瘀方组大鼠分别于5、7、9月龄（即造模术后4、6、8月）开始用药，连续用药1个月。3组大鼠分别于6、8、10月龄（即造模术后5、7、9月）处死，每次10只。处死后取大鼠膝关节，行番红O／固绿染色观察关节形态学变化，实时荧光定量聚合酶链式反应（polymerase chain reaction，PCR）检测各组关节软骨内Ⅱ型胶原基因（type Ⅱ collagen gene，Col2A1）、聚集蛋白聚糖（aggrecan-1，Agc1）、基质金属蛋白酶-13（matrix metalloproteinase-13，MMP-13）以及金属蛋白酶组织抑制剂-1（tissue inhibitor of matrix metalloproteinase-1，TIMP-1）mRNA表达的情况。 结果：模型组关节软骨结构破坏严重，而益气化瘀方组大鼠关节软骨退变较模型组明显减轻。实时荧光定量PCR结果显示，益气化瘀方组6和10月龄大鼠膝关节软骨Col2A1、Agc1以及TIMP-1 mRNA表达量高于模型组（P〈0．01，P〈0．05），MMP-13 mRNA表达与模型组比较差异无统计学意义；益气化瘀方组8月龄大鼠Agcl mRNA表达明显高于模型组（P〈0。01），MMP-13 mRNA表达明显低于模型组（P〈0．01）。 结论：益气化瘀方可通过促进软骨合成聚集蛋白聚糖与Ⅱ型胶原，延缓膝骨关节炎大鼠关节软骨的退变。

Preventive and therapeutic effects of Yiqi Huayu Recipe on degeneration of articular cartilage in rats with osteoarthritis

Objective： To study the efficacy and possible mechanism of Yiqi Huayu Recipe （YQHYR）, a compound traditional Chinese herbal medicine, in preventing and treating degeneration of the articular cartilage in rats with osteoarthritis （OA）.
Methods. A total of 90 one-month-old SD rats were randomly divided into normal control group, untreated group and YQHYR group, with 30 rats in each group. The osteoarthritis was induced by shoulder disarticulation plus upright posture in rats. The rats in YQHYR group were treated with YQHYR at 5-, 7- and 9-month old （4, 6 and 8 months after the surgery） for one month. Ten rats in each subgroup were sacrificed at 6-, 8- and 10-month old （5, 7 and 9 months after the surgery） respectively and the knee joint samples were harvested for detection. Safranin-O/fast green staining was performed to examine the morphology of articular cartilage. The expressions of type Ⅱ collagen （Co12A1）, aggrecan-1 （Agc1）, matrix metalloproteinase-13 （MMP-13） and tissue inhibitor of metalloproteinase-1 （TIMP-1） mRNAs were evaluated by real-time fluorescent quantitation polymerase chain reaction. Results： Histological analysis showed that the structure of articular cartilage was seriously destroyed in rats in the untreated group. On the contrary, the degenerative changes of the articular cartilage in the YQHYR group were dropped off. Real-time fluorescent quantitation polymerase chain reaction showed that expressions of Agc1, TIMP-1 and Co12A1 mRNAs were up-regulated in 6- and 10-month-old rats in the YQHYR group as compared with those in the untreated group （P〈0.01, P〈0.05）, but there was no significant difference in expression of MMP-13 mRNA between the YQHYR group and the untreated group. The expression of Agcl mRNA was up-regulated and the expression of MMP-]3 mRNA was down-regulated in the 8-month-old rats in YQHYR group as compared with those in the untreated group （P〈0.01）.
Conclusion： YQHYR can promote the synthesizing of aggrecan and type Ⅱ collagen in chondrocytes and delay articular cartilage degradation in OA rats.