摘要
背景:膝骨关节炎是临床常见退行性关节疾病,主要表现为慢性炎症和氧化应激反应。白藜芦醇具有抗炎和抗氧化应激的生物作用,因此可对症进行治疗,有望为膝骨关节炎治疗提供新的策略。目的:探讨白藜芦醇通过沉默信息调节因子1(silence information regulator 1,SIRT1)/叉头框转录因子O1(forkhead transcription factor O1,FOXO1)通路对大鼠膝骨关节炎的作用机制。方法:40只SD大鼠随机分为对照组、模型组、白藜芦醇低剂量组和白藜芦醇高剂量组,每组10只,除对照组外均建立膝骨关节炎模型,分别在造模的第1,4,7天注射4%木瓜蛋白酶溶液与0.3 mol/L半胱氨酸溶液(1∶1混置0.5 h)的混合液20μL。造模成功后1 d开始给药,白藜芦醇低剂量组和白藜芦醇高剂量组大鼠经关节腔注射白藜芦醇25 mg/kg或100 mg/kg,对照组和模型组经关节腔注射等体积生理盐水。治疗28 d后,测定4组大鼠膝关节最大活动度;应用放射免疫法和ELISA法检测膝关节液中氧化应激指标和炎症因子水平;番红O-固绿染色分析膝关节中胶原纤维含量;采用Mankin组织学评分分析关节炎病变程度;蛋白质免疫印迹法检测膝关节SIRT1和FOXO1水平。结果与结论:①与模型组比较,白藜芦醇低剂量组和高剂量组大鼠膝关节最大屈伸角度明显增加,且高剂量组明显高于低剂量组(P<0.05);②与模型组比较,白藜芦醇低剂量组和高剂量组大鼠膝关节液中超氧化物歧化酶和谷胱甘肽过氧化物酶水平显著增加;而丙二醛水平显著下降,且高剂量组明显优于低剂量组(P<0.05);③与模型组比较,白藜芦醇低剂量组和高剂量组大鼠膝关节液炎症因子白细胞介素1β、白细胞介素6和肿瘤坏死因子α水平明显下降,且高剂量组明显低于低剂量组(P<0.05);④与模型组比较,白藜芦醇低剂量组和高剂量组大鼠膝关节胶原纤维含量增加,且高剂量组明显高于低剂量组(P<0.05);⑤与模型组比较,白藜芦醇低剂量组和高剂量组大鼠膝关节SIRT1表达水平显著增加,乙酰化FOXO1水平显著下调,且高剂量组变化幅度明显优于低剂量组(P<0.05);⑥提示白藜芦醇可显著改善膝骨关节炎大鼠关节液中的氧化应激和炎症因子水平,缓解关节炎症状,且呈剂量依赖性,可能是通过SIRT1/FOXO1通路实现的。
BACKGROUND:Knee osteoarthritis is a common clinical degenerative joint disease characterized by chronic inflammation and oxidative stress.Resveratrol has anti-inflammatory and anti-oxidative stress biological effects,and therefore it can be used symptomatically and expected to provide a new strategy for the treatment of knee osteoarthritis.OBJECTIVE:To investigate the therapeutic effect and mechanism of resveratrol on knee osteoarthritis in rats through the silence information regulator 1(SIRT1)/forkhead transcription factor O1(FOXO1)pathway.METHODS:Forty Sprague-Dawley rats were randomly divided into control group,model group,low-dose resveratrol group,and high-dose resveratrol group,with 10 rats in each group.Knee osteoarthritis models were established in the model group,low-dose resveratrol group,and high-dose resveratrol group.A mixture of 4%papain solution and 0.3 mol/L cysteine solution(1:1 for 0.5 hours;20μL)was injected at 1,4,and 7 days after modeling.Rats in the lowdose and high-dose resveratrol groups were injected with 25 and 100 mg/kg resveratrol through the articular cavity at 1 day after successful modeling,while those in the control and model groups were injected with equivalent volume of physiological saline through the articular cavity.After 28 days of treatment,the maximum knee joint activity was measured;the levels of oxidative stress indicators and inflammatory factors in the synovial fluid of the knee joint were analyzed by radioimmunoassay and ELISA;the content of collagen fibers in the knee joint was analyzed by safranin O-fast green staining;the degree of arthritic lesions was analyzed using the Mankin histological score;and the levels of SIRT1 and FOXO1 in the knee joint were detected by western blot assay.RESULTS AND CONCLUSION:Compared with the model group,the maximum knee flexion and extension angles of rats significantly increased in the low-dose and high-dose resveratrol groups,and were significantly higher in the high-dose group than the low-dose group(P<0.05).Compared with the model group,the levels of superoxide dismutase and glutathione peroxidase in the knee joint fluid of rats significantly increased in the low-dose and high-dose resveratrol groups.The level of malondialdehyde significantly decreased in both resveratrol groups,and the level in the high-dose resveratrol group was significantly better than that in the low-dose resveratrol group(P<0.05).Compared with the model group,the low-dose and high-dose resveratrol groups showed a significant decrease in the levels of interleukin 1β,interleukin 6 and tumor necrosis factorαin the knee joint fluid of rats,and the levels of these inflammatory factors were significantly lower in the high-dose resveratrol group than the low-dose resveratrol group(P<0.05).Compared with the model group,the content of collagen fibers in the knee joint was significantly increased in both resveratrol groups,and the high-dose resveratrol group showed a higher content of collagen fibers than the low-dose resveratrol group(P<0.05).Compared with the model group,the expression level of SIRT1 in the knee joints of rats significantly increased in both resveratrol groups,while the level of acetylated FOXO1 significantly decreased(P<0.05).The magnitude of changes was significantly better in the high-dose group than the low-dose group.To conclude,resveratrol significantly improves the levels of oxidative stress and inflammatory factors in the joint fluid of rats with knee osteoarthritis and alleviates arthritic symptoms in a dose-dependent manner,possibly through the SIRT1/FOXO1 pathway.
作者
任伟亮
焦永伟
张健
杨立英
杨琦
Ren Weiliang;Jiao Yongwei;Zhang Jian;Yang Liying;Yang Qi(Department of Orthopedics and Traumatology,The First Affiliated Hospital of Hebei University of Chinese Medicine(Hebei Provincial Hospital of Traditional Chinese Medicine),Shijiazhuang 050000,Hebei Province,China)
出处
《中国组织工程研究》
CAS
北大核心
2024年第32期5154-5158,共5页
Chinese Journal of Tissue Engineering Research
基金
2020年度河北省中医药类科研计划项目(2020008),项目负责人:任伟亮。
