葡萄籽原花青素提取物对兔激素性股骨头坏死的保护作用及机制研究

Mechanism of grape seed procyanidins extract in protecting steroid-induced femoral head necrosis

目的探讨葡萄籽原花青素提取物(GSPE)对兔激素性股骨头坏死模型的保护作用及其相关机制。方法 50只新西兰大白兔随机分为3组,模型组20只,干预组20只,对照组10只。模型组肌肉注射单次剂量(40 mg/kg)甲基强的松龙,干预组在注射甲基强的松龙的同时加用GSPE(200 mg/kg)灌胃,对照组肌肉注射与甲基强的松龙等量的生理盐水。2周后所有兔被处死并收集双侧股骨头组织,采用HE染色检测骨坏死发生率;采用免疫组化检测DNA损坏标志物8-OHdG,骨组织中SOD、GSH-Px活性及MDA水平;通过Tunel染色和检测Casepase-3活性观察凋亡情况。结果 7只兔死亡,其中模型组4只,干预组3只。干预组股骨头坏死发生率、8-OHdG比例明显低于模型组,差异有统计学意义(P <0.05)。干预组股骨头组织SOD活性、GSH-Px高于模型组,MDA含量低于模型组,差异有统计学意义(P <0.05)。模型组凋亡细胞数量明显增多,而干预组后凋亡细胞明显减少。模型组Caspase 3活性明显上升,干预组Caspase 3活性明显降低。结论 GSPE通过抗氧化应激和抗凋亡作用对兔激素性股骨头坏死模型起到保护作用。

Objective To explore the protective effect of grape seed procyanidins extract(GSPE) on rabbit steroid-induced osteonecrosis model and its related mechanism. Methods Fifty New Zealand white rabbits were randomly divided into 3 groups, model group(steroid alone, n =20), intervention group(steroid+GSPE, n =20) and control group(physiological saline only, n =10). In the model group, a single dose of methylprednisolone(40 mg/kg) was used. In the intervention group,methylprednisolone was applied and GSPE(200 mg/kg) was added every day until death. The control group was intramusculally injected with the same amount of normal saline as methylprednisolone. After two weeks, all the animals were executed and bilateral femoral head tissues were collected. HE staining was used to detect the incidence of osteonecrosis.DNA damage markers, 8-OHdG was detected by immunohistochemistry, while super oxide dismutase(SOD), glutathione peroxidase(Gsh-px) activity and malondialdehyde(MDA) level were also detected. Apoptosis was observed by Tunel staining and Casepase-3 activity. Results Seven rabbits died, including four in the model group and three in the intervention group.The incidence of femoral head necrosis and the ratio of 8-OHdG in the intervention group were significantly lower than those in the model group(P <0.05). The activity of SOD and GSH-Px in femoral head tissue of intervention group were higher than that of model group, and the content of MDA was lower than that of model group(P <0.05). The number of apoptotic cells increased significantly in the model group, but decreased significantly in the intervention group. The activity of Caspase 3 in model group increased significantly, while that in intervention group decreased significantly. Conclusion GSPE has protective effects on rabbit steroid-induced osteonecrosis model by anti-oxidative stress and anti-apoptosis.