来源：风湿病与关节炎,2018,7（6）:5-9,14.

狗脊多糖对硝普钠诱导退变大鼠软骨细胞氧自由基影响的研究（临床研究）

付长龙^1,2，梅阳阳³，李　民^1,2，黄志强⁴，黄艳峰⁴，林　晴⁴，郑春松⁴，叶锦霞⁴

　　【摘　要】目的：观察狗脊多糖对硝普钠（SNP）诱导退变大鼠软骨细胞氧自由基的影响，为其在骨关节炎的临床应用提供理论依据。方法：采用水提醇沉法、Sevag法除蛋白提取纯化获得狗脊多糖，苯酚-硫酸比色法测定其多糖含量；酶联免疫吸附测定法分别检测体外培养大鼠软骨细胞空白对照组、模型对照组、各剂量狗脊多糖组（100，200，400，800 μg·mL^-1）的超氧化物歧化酶（SOD）、一氧化氮（NO）、丙二醛（MDA）的含量。结果：狗脊多糖含量为76.91%，精密度试验RSD = 1.86%，表明仪器精密度良好，且稳定性试验RSD = 1.98%，平均回收率试验为95.4%，RSD = 1.83%；酶联免疫吸附法检测经SNP造模后软骨细胞氧自由基表达结果显示，与空白对照组比较，模型对照组SOD、NO、MDA含量表达差异均有统计学意义（P < 0.01），提示软骨细胞退变模型建立；与模型对照组比较，各剂量狗脊多糖组（100，200，400，800 μg·mL^-1）干预48 h后，可上调SOD含量，下调NO、MDA含量，其中以200 μg·mL^-1组表达最显著（P < 0.01）。结论：苯酚-硫酸比色法可作为狗脊多糖的含量测定方法；狗脊多糖可降低经SNP造模后软骨细胞中异常氧自由基含量表达，上调抗氧化因子水平，发挥抗氧化作用。

　　【关键词】　骨关节炎，膝；狗脊多糖；软骨细胞；氧自由基；大鼠

Effects of Rhizoma Cibotii Polysaccharide on Oxygen Free Radicals in Rats' Degenerative Chondrocytes Induced by Sodium Nitroprusside

FU Chang-long,MEI Yang-yang,LI Min,HUANG Zhi-qiang,HUANG Yan-feng,LIN Qing,ZHENG Chun-song,YE Jin-xia

　　【ABSTRACT】Objective:To observe the effects of Rhizoma Cibotii polysaccharides on oxygen free radicals in rats' degenerative chondrocytes induced by sodium nitroprusside（SNP）and provide a theoretical basis for its clinical application in osteoarthritis.Methods:Polysaccharide was extracted and purified with water extraction and Sevag methods,and the polysaccharide content was determined by the phenol sulfuric acid colorimetry.The enzyme linked immunosorbent assay was used to detect the contents of superoxide dismutase（SOD）,nitric oxide（NO）and malondialdehyde（MDA）respectively in the blank group,the model group and the Rhizoma Cibotii polysaccharides group（100,200,400,800 μg·mL^-1）.Results:The Rhizoma Cibotii polysaccharide content was 76.91% and the precision experiment showed RSD = 1.86%,which showed that the precision of the instrument was good.The stability test showed RSD = 1.98%,the average recovery test was 95.4%,and RSD was 1.83%.The enzyme linked immunosorbent assay showed that the differences between the contents of SOD,NO and MDA in the model group and thosein the blank group were statistically significant（P < 0.01）,suggesting that the chond-rocyte degeneration model was established.Compared with the model group,after the intervention to the experimental group（100,200,400,800 μg·mL^-1）for 48 h,the contents of SOD could be up regulated and the content of NO and MDA could be down regulated,with the group of 200 µg·mL^-1 the most significant（P < 0.01）.Conclusion:The phenol sulfuric acid colorimetry can be used to determine the content of the Rhizoma Cibotii polysaccharide.Rhizoma Cibotii polysaccharide can reduce the expression of the abnormal oxygen free radical content in the chondrocytes after the SNP model,up-regulating the level of antioxidant factors and playing an antioxidant role.

　　【Keywords】 osteoarthritis,knee;Rhizoma Cibotii polysaccharide;chondrocytes;oxygen free radicals;rats