葡萄籽原花青素提取物对运动性骨关节损伤大鼠的影响及其作用机制的研究

Effects of grape seed proanthocyanidin extract on sports joint injury in rats

目的揭示葡萄籽原花青素提取物(GSPE)对大鼠运动性骨关节损伤(SJI)的治疗作用及机制。方法建立SJI大鼠模型后,各组大鼠经口给予不同剂量的GSPE(0、50、100和200 mg/kg),持续4周,并设立正常对照组。受试物给药结束后,通过机械痛阈(PWT)测定法测定各组模型大鼠疼痛情况;取尾静脉血,检测血清超氧化物歧化酶(SOD)、谷胱甘肽过氧化物酶(GSH-Px)、丙二醛(MDA)含量。处死后,取膝关节经番红O-固绿染色后进行病理检查;取膝关节组织,通过Western blot方法检测过氧化物酶体增殖物激活受体γ共激活因子α(PGC-1α)、磷酸化-AMP依赖的蛋白激酶α(p-AMPKα)、AMPKα、核呼吸因子1(NRF1)和线粒体转录因子A(TFAM)的蛋白表达水平。结果随着GSPE剂量增加,SJI大鼠的PWT显著上升(P<0.05),膝关节病变形态改善,SJI大鼠的血清SOD和GSH-Px含量升高,MDA含量显著降低(P<0.05)。SJI大鼠膝关节组织中PGC-1α、p-AMPKα/AMPKα、NRF-1和TFAM的蛋白表达水平随着GSPE剂量增加而提高(P<0.05)。结论GSPE摄入有效减轻了SJI大鼠疼痛和膝关节损伤,作用机制与AMPK/PGC-1α线粒体通路的激活抑制了体内氧化应激水平有关。

Objective To reveal the therapeutic effect and mechanism of grape seed proanthocyanidin extract(GSPE)on sports joint injury(SJI)in rats.Methods After the establishment of the SJI rat model,rats in each group were orally administered different doses of GSPE(0,50,100,and 200 mg/kg)for 4 weeks,and a normal control group was established.After the administration of the test substance,the pain condition of the model rats in each group was assessed using the paw withdrawal threshold(PWT)assay.Tail vein blood was collected to detect serum superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),and malondialdehyde(MDA)content.After the rats were sacrificed,the knee joints were harvested and stained with safranin O-fast green for pathological examination.The protein expression levels of peroxisome proliferator-activated receptor gamma coactivator alpha(PGC-1α),phosphorylated-AMP-dependent protein kinase alpha(p-AMPKα),AMPKα,nuclear respiratory factor 1(NRF1),and mitochondrial transcription factor A(TFAM)in knee joint tissue were determined using Western blot.Results With increasing GSPE dose,the PWT of rats with SJI increased(P<0.05),the shape of knee joint lesions improved,serum SOD and GSH-Px contents increased,and MDA content decreased(P<0.05).The protein expression levels of PGC-1α,p-AMPKα/AMPKα,NRF-1,and TFAM in knee joint tissues of rats with SJI increased with increasing GSPE dose(P<0.05).Conclusion GSPE intake effectively alleviated pain and knee joint injury in rats with SJI,and the mechanism was related to the activation of the AMPK/PGC-1αmitochondrial pathway,which inhibits oxidative stress in vivo.