芸香苷改善大鼠椎间盘退变的机制  

Rutin attenuates the progression of intervertebral disc degeneration in rats

作　　者：许大勇[1] 李云朋[1] 魏景梅[1] 刘汝银[1] Xu Dayong;Li Yunpeng;Wei Jingmei;Liu Ruyin(Department of Spine Surgery,Henan Hospital of Traditional Chinese Medicine,Zhengzhou 450000,Henan Province,China)

机构地区：[1]河南省中医院脊柱外科,河南省郑州市450000

出　　处：《中国组织工程研究》2023年第14期2139-2145,共7页Chinese Journal of Tissue Engineering Research

基　　金：河南省中医药拔尖人才培养项目资助课题(2019ZYBJ18),项目负责人:刘汝银。

摘　　要：背景:芸香苷具有抗氧化、抗炎、抗细胞凋亡、血管保护、神经保护和心脏保护等药理活性,可用于治疗癌症、糖尿病、高血压和高胆固醇血症等多种疾病,但其在椎间盘退变性疾病中的研究较少。目的:探究芸香苷对大鼠椎间盘退变的调控和作用机制。方法:(1)从大鼠髓核组织中分离获得髓核细胞,之后用芸香苷(10,50,100,200,400μmol/L)和白细胞介素1β(10μg/L)处理髓核细胞,检测细胞活性、炎症因子和氧化应激因子的表达以及细胞代谢酶的生成、基质降解和细胞凋亡的情况;之后加入WNT/β-catenin通路激活剂R-spondin1处理髓核细胞,检测β-catenin、c-Myc和Cyclin D1表达。(2)36只SD大鼠随机选取其中26只,用针刺纤维环构建椎间盘退变模型,1周后通过磁共振成像(MRI)进行评估,其余10只为假手术组做对照。建模成功的大鼠随机分为椎间盘退变模型组和芸香苷组,芸香苷组大鼠使用40 mg/kg芸香苷灌胃处理8周,模型组和假手术组大鼠用等量的生理盐水处理。Pfirrmann分级系统评定椎间盘退变程度;ELISA检测各组大鼠血清中白细胞介素6,胶原Ⅱ浓度和caspase-3活性;Western blotting检测各组大鼠髓核组织中聚集蛋白聚糖和诱导型一氧化氮合酶的蛋白表达水平。结果与结论:(1)细胞实验:100μmol/L芸香苷能够完全缓解白细胞介素1β造成的髓核细胞毒性,抑制白细胞介素6、肿瘤坏死因子α、环氧化酶2、诱导型一氧化氮合酶、基质金属蛋白酶3、13和ADAMTS-5表达,抑制胶原Ⅱ、聚集蛋白聚糖降解和细胞凋亡,R-spondin1能逆转芸香苷的作用;(2)体内研究发现,与椎间盘退变模型组相比,芸香苷组的大鼠血清中白细胞介素6质量浓度降低,胶原Ⅱ质量浓度升高,caspase-3活性降低,髓核组织中诱导型一氧化氮合酶蛋白水平下调,聚集蛋白聚糖蛋白水平升高,椎间盘退变有明显的改善;(3)结果说明,芸香苷通过抗炎、抗氧化�BACKGROUND:Rutin has various pharmacological activities including anti-oxidant,anti-inflammatory,anti-apoptottc,vasoprotective,n euro protective,and cardioprotective properties,which can be used to treat diseases such as cance r,diabetes,hype rtension,and hypercholesterolemia.Howeve r,it has been less studied in the treatment of intervertebral disc degenerative diseases.OBJECTIVE:To investigate the regulatory effect of rutin on intervertebral disc degeneration in rats and its mechanism.METHODS:Nucleus pulposus cells were isolated from the postnatal rat nucleus pulposus tissue and then treated with rutin(10,50,100,200,or 400 μmol/L)and inte rleukin-1β(10μg/L).Cell viability and expression of inflammatory factor and oxida tive stress factors were detected.P roduction of cellular metabolic enzymes,matrix degradation,and cell apoptosis were also observed.Furthermore,treatment of nucleus pulposus cells with R-spondin1,a WNT/β-catenin pathway activator,and the expression of β-catenin,c-Myc,and Cyclin D1 were detected.The 26 out of 36 Sprague-Dawley rats were randomly selected,to construct animal models of intervertebral disc degeneration using a needle fiber loop.One week late r,the model was evaluated by magnetic resonance imaging.The rest 10 rats were used as controls.Rats with successful modeling were randomly divided into a model group and a rutin group.Rats in the rutin group were intragastrically treated with 40 mg/kg rutin for 8 wee ks,while those in the model group were treated with an equal volume of normal saline.The Pfirrmann grading system was used to grade the degree of interve rtebral disc degeneration.The serum levels of interleukin-6,type Ⅱ collagen and caspase-3 were detected using ELISA.The protein levels of Aggrecan and inducible nitric oxide synthase in rat nucleus pulpos us tissue were measured using western blot assay.RESULTS AND CONCLUSION:(1) Cell expe riment:100 μmol/L ru tin completely alleviated the toxicity of interleukin-1β to nucleus pulpos us cells,decreased the expression of int

关 键 词：椎间盘退变 芸香苷 抗氧化 抗炎 WNT/Β-CATENIN信号通路 

分 类 号：R446[医药卫生—诊断学] R496[医药卫生—临床医学] R318