壮骨1号方对大鼠骨性关节炎的抗损伤作用

Study on anti-injury effect and pathomorphology changes of Zhuangguyihao Fang for osteoarthritis

目的:观察壮骨1号方对大鼠骨性关节炎的抗损伤作用及病理形态学的影响。方法:SD大鼠,以0.05ml(1.6U)/关节剂量,双侧膝关节注射4%木瓜蛋白酶-泊洛沙姆温敏型原位凝胶控释剂复制大鼠骨性关节炎模型。选成功模型鼠48只,随机分为骨性关节炎模型组、壮骨1号方2.20 g/kg、1.10g/kg、0.55 g/kg组、壮骨关节丸组和氨基葡萄糖组,另设正常对照组,每组8只,按设定剂量每天灌胃1次,连续治疗8周。测定关节液前列腺素E2(PGE2)和肿瘤坏死因子α(TNF-α)含量;测定关节软骨诱导型一氧化氮合成酶(iNOs)mRNA、转化生长因子β1(TGF-β1)mRNA表达;观察关节软骨病理组织学变化并进行病理损伤评价。结果:模型组大鼠关节液PGE2和TNF-α含量明显升高(P<0.01),治疗8周后,壮骨1号方2.20、1.10 g/kg组关节液PGE2和TNF-α含量明显降低(P<0.01)。模型组大鼠软骨iNOs mRNA、TGF-β1 mRNA表达明显增强(P<0.05或P<0.01),壮骨1号方2.20,1.10,0.55 g/kg治疗8周后,iNOs mRNA、TGF-β1 mRNA表达均减弱(P<0.05或P<0.01)。模型组大鼠关节软骨病理损伤明显,病理总评分为2.75±1.03,用壮骨1号方2.20,1.10g/kg治疗8周后,关节软骨损伤明显减轻,病理总评分分别为1.62±0.91和1.87±0.64(P<0.05或P<0.01)。结论:壮骨1号方具有一定的抗软骨损伤作用,能延缓软骨退行性变的进程,这种抗损伤作用可能与通过壮骨1号方的补益肝肾功能,调节PGE2、TNF-α、iNOS、TGF-β1的作用相关。

Objective： To explore the anti-injury effect and pathomorphology changes of Zhuangguyihao Fang for osteoarthritis（ OA）. Methods：healthy male SD rats were selected and the OA rat model was induced by sustained releasing agent（ 4% Papain and poloxamer）. 0. 05ml of the sustained releasing agent（ 4% Papain and poloxamer） was injected into both knee joint compartments of the rats to injury cartilage for a period of 2h at 50℃. 48 of rats with established osteoarthritis were randomly divided into 6 groups： model group,Zhuangguyihao Fang（ 2.20,1. 10,0. 55 g /kg） groups,Zhuanggu Guanjie Pill（ 1. 10 g /kg） group and glucosamine group（ 0. 03 g /kg）. The numbers of rats in each group were 8. In addition,8 healthy rats were selected as normal group. The drugs mentioned above were administered to the rats with established osteoarthritis by gastric perfusion once a day for 8 weeks. Isovolumic distilled water was administered to the normal group and the model group by gastric perfusion once a day for 8 weeks. The contents of PGE2and TNF-α in synovial fluid were detected. The expressions of iNOS and TGF-β1 mRNA in cartilages were verified and the changes in cartilages were examined and histopathology evaluations were conducted. Results： Compared with the normal group,the contents of PGE2and TNF-α in the model group was significantly increased（ P ＆lt; 0. 01）.After 8 weeks’ treatment,compared with the model group,Zhuangguyihao Fang（ 1. 10,2. 20 g /kg） treatments significantly reduced the contents of PGE2and TNF-α（ P ＆lt; 0. 01）. Compared with the normal group,the expressions of iNOS and TGF-β1 mRNA in cartilages of the model group were up-regulated（ P ＆lt; 0. 05,P ＆lt; 0. 01）. Compared with the model group,Zhuangguyihao Fang（ 0. 55,1. 10,2. 20 g /kg） treatments significantly down-regulated the expressions of iNOS and TGF-β1 mRNA in cartilages（ P ＆lt; 0. 05,P ＆lt; 0. 01）. Apparent pathology lesions had been shown in both knee articular cartilages of model group,the grading score of cartilage histopathology was 2. 75 ± 1. 03. Zhuangguyihao Fang（ 1. 10 and 2. 20 g /kg respectively） treatments significantly reduced pathology lesions in cartilages and the grading scores of cartilage histopathology,which were 1. 62 ± 0. 91 and 1. 87 ± 0. 64 respectively（ P ＆lt; 0. 05,P ＆lt; 0. 01）. Conclusion： Zhuangguyihao Fang has been shown to exhibit the anti-injury effect in cartilage and attenuate the progression of OA,which is related to the regulation of PGE2,TNF-α,iNOS and TGF-β1.