白藜芦醇对兔激素诱导股骨头缺血性坏死抗氧化应激和抗凋亡作用的实验研究

Anti-oxidative stress and anti-apoptotic effects of resveratrol on steroid-induced femoral head necrosis in rabbits

目的探讨白藜芦醇对兔激素诱导股骨头缺血性坏死的抗氧化应激和抗凋亡作用。方法将60只新西兰白兔随机分为3组,模型组24只,干预组24只,对照组12只。模型组第1天静脉注射1次脂多糖(4 mg/kg),从第2天开始连续3 d肌肉注射甲基强的松龙(60 mg/kg,每天1次);干预组第1天静脉注射1次脂多糖(4 mg/kg),从第2天开始连续3 d肌肉注射甲基强的松龙(60 mg/kg,每天1次),并且从第1天开始每天口服1次白藜芦醇(20 mg/kg),持续2周;对照组从第1天开始肌肉注射等量生理盐水。2周后处死新西兰白兔并收集双侧股骨头组织,通过测定骨组织抗氧化酶活性和丙二醛水平以观察氧化应激情况,进行Tunel染色及Caspase 3活性检测观察细胞凋亡情况。结果股骨头组织超氧化物歧化酶、谷胱甘肽过氧化物酶活性检测结果显示,对照组的活性最强,模型组的活性最低,干预组的活性居中,组间差异有统计学意义(P<0.05)。3组间股骨头组织丙二醛含量差异有统计学意义(P<0.05),对照组最低,模型组最高,干预组居中。Tunel染色结果显示,干预组凋亡细胞数与模型组相比明显减少。模型组凋亡指数较对照组显著增加,而干预组凋亡指数较模型组明显降低;模型组Caspase 3活性较对照组升高,而干预组Caspase 3活性较模型组降低,差异有统计学意义(P<0.05)。结论对于兔激素性股骨头缺血性坏死,白藜芦醇可通过发挥其抗氧化应激和抗凋亡特性起到一定的预防和治疗作用,有望发展其成为保护股骨头的临床药物。

Objective To investigate the anti-oxidative stress and anti-apoptotic effects of resveratrol on steroid-induced femoral head necrosis in rabbits.Methods Sixty New Zealand white rabbits were randomly divided into three groups,24 in the model group,24 in the intervention group,and 12 in the control group.In the model group,lipopolysaccharide(4 mg/kg)was administered intravenously once on day 1,and methylprednisolone(60 mg/kg,once a day)was administered intramuscularly for 3consecutive days from day 2.In the intervention group,lipopolysaccharide(4 mg/kg)was administered intravenously once on day 1,methylprednisolone was administered intramuscularly(60 mg/kg,once a day)from day 2 for 3 consecutive days,and resveratrol was administered orally once a day from day 1(20 mg/kg,once a day).The control group received an intramuscular injection of equal volume of normal saline from day 1.New Zealand white rabbits were sacrificed after 2 weeks and bilateral femoral head tissues were collected,determining bone tissue antioxidant enzyme activities and malondialdehyde levels to observe oxidative stress conditions,and performing Tunel staining as well as caspase 3 activity detection to observe apoptosis.Results The activities of superoxide dismutase and glutathione peroxidase in femoral head tissues were detected,and the results showed that the control group had the strongest activity,the model group had the lowest activity,and the intervention group had a medium activity,and the differences between groups were statistically significant(P<0.05).There was a significant difference in the malondialdehyde content of femoral head tissues among the 3 groups(P<0.05),with the lowest in the control group,the highest in the model group,and the medium in the intervention group.Tunel staining results showed that the number of apoptotic cells was significantly reduced in the intervention group compared with the model group.The apoptotic index was significantly increased in the model group compared with the control group,whereas it decreased in the intervention group compared with the model group;Caspase 3 activity in the model group was significantly higher than that in the control group,and caspase 3 activity in the intervention group was significantly lower than that in the model group,with a significant difference(P<0.05).Conclusion For steroid-induced avascular necrosis of the femoral head in rabbits,resveratrol may play a certain preventive and therapeutic role by exerting its anti-oxidative stress and anti-apoptotic properties,which is expected to further develop it into a clinical drug for protecting the femoral head.