补肾活血汤干预骨质疏松并骨关节炎模型兔软骨多种细胞因子mRNA的表达

Bushen Huoxue Decoction affects mRNA expression of various cytokines in cartilage of the rabbit models of osteoporotic osteoarthritis

背景:临床疗效观察发现补肾活血汤对骨质疏松与骨关节炎均有良好的临床疗效,但关于其作用机制研究较少。目的:探讨补肾活血汤对骨质疏松并骨关节炎新西兰雌性大白兔模型膝部软骨细胞因子白细胞介素1、白细胞介素6、胰岛素样生长因子1、转化生长因子βmRNA表达的影响。方法:(1)30只新西兰雌性大白兔被随机分为骨质疏松症组、骨性关节炎组及骨质疏松症并骨性关节炎组(OA+OP组),造模成功后均提取兔右膝关节软骨组织并分离培养软骨细胞;(2)另取7只兔制备7组含药血清:正常血清组、低剂量(补肾活血汤)组、中剂量(补肾活血汤)组、高剂量(补肾活血汤)组、盐酸氨基葡萄糖组、阿仑膦酸钠组、盐酸氨基葡萄糖+阿仑磷酸钠组;(3)MTT法观察7组含药血清对兔软骨细胞增殖的影响;PCR法检测7组药物对兔软骨细胞中细胞因子白细胞介素1、白细胞介素6、胰岛素样生长因子1、转化生长因子βmRNA表达的影响。结果与结论:(1)MTT检测结果显示:随着药物作用时间的延长至第7天,除骨性关节炎组低剂量组外,其他各剂量组与正常组对比吸光度值均增高(P <0.05),3组呈剂量-时间-效益关系;(2)PCR结果显示:软骨细胞中白细胞介素1mRNA表达量OA+OP组>骨性关节炎组>骨质疏松症组(P<0.05);白细胞介素6mRNA表达量OA+OP组>骨性关节炎组>骨质疏松症组,骨质疏松症组与OA+OP组及骨性关节炎组对比,差异有显著性意义(P<0.05);胰岛素样生长因子1、转化生长因子βmRNA表达量骨质疏松症组>骨性关节炎组>OA+OP组,骨质疏松症组与OA+OP组对比差异有显著性意义(P <0.05);与正常组比较,各剂量组均有降低软骨细胞内白细胞介素1、白细胞介素6 mRNA表达及升高胰岛素样生长因子1、转化生长因子βmRNA表达的作用,呈现剂量-效益关系,以高剂量组作用最明显;(3)结果说明,补肾活血汤有促软骨细胞增殖的作用;骨性关节炎和骨质疏松症的发病与白细胞介素1、白细胞介素6、胰岛素样生长因子1、转化生长因子βmRNA表达相关,骨性关节炎与骨质疏松症呈正相关关系;补肾活血汤能间接或直接保护及修复软骨细胞。

BACKGROUND:Bushen Huoxue Decoction has been found to be effective for osteoarthritis and osteoporosis,but the underlying mechanisms are little reported.OBJECTIVE:To explore the effects of Bushen Huoxue Decoction on the mRNA expression levels of interleukin 1,interleukin 6,and transforming growth factor-βin the knee chondrocytes of female New Zealand female rabbit models of osteoporotic osteoarthritis.METHODS:Thirty female New Zealand rabbits were randomly divided into osteoporosis,osteoarthritis,and osteoporotic osteoarthritis groups.The right knee articular cartilages were removed,and the chondrocytes were isolated and cultured after modeling.Another seven rabbits were selected to prepare seven groups of serum containing drugs:normal group,low-,medium-and high-dose Bushen Huoxue Decoction groups,glucosamine hydrochloride group,alendronate sodium group and glucosamine hydrochloride plus alendronate sodium group.The chondrocyte proliferation in each group was detected by MTT assay.The mRNA expression levels of interleukin 1,interleukin 6,insulin-like growth factor-1(IGF-1)and transforming growth factor-β(TGF-β)in the knee chondrocytes were detected by PCR.RESULTS AND CONCLUSION:MTT results showed that with the administration time increasing to day 7,the absorbance values in each group were significantly higher than those in the normal group(P<0.05),except osteoarthritis and low-dose groups.Bushen Huoxue Decoction exerted effect in a concentration-dependent manner.PCR results:the order of mRNA expression level of interleukin 1 was as follows:osteoporotic osteoarthritis group>osteoarthritis group>osteoporosis group(P<0.05).The mRNA expression level of interleukin 6 was the highest in the osteoporotic osteoarthritis group,followed by osteoarthritis group,the lowest in the osteoporosis group,and the level in the osteoporotic osteoarthritis and osteoarthritis groups was significantly higher than that in the osteoporosis group(P<0.05).The order of mRNA expression levels of IGF-1 and TGF-βwere the highest in the osteoporosis group,followed by osteoarthritis group,the lowest in the osteoporotic osteoarthritis group,and the difference was significant between osteoporosis and osteoporotic osteoarthritis groups(P<0.05).Compared with the normal group,Bushen Huoxue Decoction significantly decreased the mRNA expression levels of interleukin 1 and interleukin 6,and significantly increased the mRNA expression levels of IGF-1 and TGF-βin a concentration-dependent manner.In summary,Bushen Huoxue Decoction can promote the proliferation of chondrocytes.The occurrence of osteoarthritis and osteoporosis is related to interleukin 1,interleukin 6,IGF-1 and TGF-βmRNA expression.There is a positive correlation between osteoarthritis and osteoporosis.Bushen Huoxue Decoction indirectly or directly protects and repairs injured chondrocytes.