刺老苞根皮黄酮对抗骨性关节炎软骨细胞氧化损伤及炎性的影响研究

Effect of Aralia echinocauIis Hand.Mazz Flavonoids against Oxidative Damage and Inflammatory of Osteoarthritis Cartilage Cells

目的研究刺老苞根皮黄酮对骨关节炎软骨细胞氧化损伤及炎性的逆转作用。方法选15只5月龄新西兰兔,采用内侧副韧带、前后交叉韧带切断并切除内侧半月板的Hulth骨性关节炎动物模型制作方法,分离培养软骨细胞,其中模型组分别加入10,20,30μg/ml刺老苞根皮黄酮低、中、高剂量处理。从第9周开始,采用DCFH-DA法检测细胞内活性氧水平,Griess法测定细胞培养上清中NO、SOD和GSH-Px含量,RT-PCR法检测各组细胞相关炎性因子IL-1β、诱导型一氧化氮合酶iNOS、转录调节因子p53基因mRNA的表达。结果与模型组比较,刺老苞根皮黄酮各剂量组细胞内活性氧水平、NaNO2含量、IL-1β/GAPDH表达水平、iNOS/GAPDH表达水平、p53/GAPDH表达水平均有所降低,且差异均有统计学意义(P<0.01),而SOD、GSH-Px均有所升高,且差异均有统计学意义(P<0.01)。结论刺老苞根皮黄酮对骨关节炎软骨细胞的氧化损伤及炎性有逆转作用,具有研发成为骨性关节炎治疗药物的潜能。

Objective To study the reverse effect of the oxidative damage and inflammatory on cartilage cells by Aralia echinocaulis Hand. Mazz flavonoids （AHF）. Methods Fifteen New Zealand white rabbits of 5 months old were selected. Animal model was estab- lished by method of Hulth osteoarthritis animal model. The anterior and posterior cruciate ligament and medial collateral ligament were cut off and medial meniscus were cut articular cartilage cell cultured in vitro. Osteoarthritis cartilage cells in the model groups were added AHF 10,20,30 μg/ml respectively. To begin with the ninth week, the reactive oxygen species level in chondrocytes were detected by DCFH -DA,the content of NO and the activity of SOD and GSH -Px in cell culture supernatant were detected by Griess method. The related expression content of IL- I[3/GAPDH, iNOS/GAPDH and p53/GAPDH was detected by RT- PCR. Re- suits Compared with the model group, the content of intracellular reactive oxygen species,the content of NaNO2 ,the content of IL - 1 β/GAPDH expression,the content of iNOS/GAPDH expression,the content of p53/GAPDH expression were decreased and showed statistically significant difference（ P 〈 0.01 ） in AHF groups with different dosages. At the same time ,the activity of SOD and GSH - Px in model groups were increased and showed statistically significant difference （P 〈 0.01 ）, compared with the model group. Con- clusion The AHF has anti - oxidative damage and anti - inflammatory effect of osteoarthritis cartilage cells within a certain dose range. It may be the main mechanism for Aralia echinocaulis Hand. Mazz flavonoids to treat osteoarthritis.