中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (20): 2979-2984.doi: 10.3969/j.issn.2095-4344.2016.20.013

• 骨组织构建 bone tissue construction • 上一篇    下一篇

慢病毒介导shRNA关节腔注射可加重骨关节炎的软骨破坏

杨二平1,彭  飞2,梁  杰1,杜远立1   

  1. 1三峡大学人民医院,宜昌市第一人民医院,湖北省宜昌市  443000;2武汉大学人民医院,湖北省武汉市  430060
  • 出版日期:2016-05-13 发布日期:2016-05-13
  • 作者简介:杨二平,男,1972年生,湖北省蕲春县人,汉族,2015年武汉大学毕业,博士,主治医师,主要从事骨关节疾病研究。
  • 基金资助:

    国家青年科学基金(61308110)

Lentivirus-induced knockdown of low density lipoprotein receptor-related protein 1 aggravates cartilage damage in a rat model of osteoarthritis

Yang Er-ping1, Peng Fei2, Liang Jie1, Du Yuan-li1   

  1. 1Three Gorges University People’s Hospital, Yichang First People’s Hospital, Yichang 443000, Hubei Province, China; 2Renmin Hospital of Wuhan University, Wuhan 430060, Hubei Province, China
  • Online:2016-05-13 Published:2016-05-13
  • About author:Yang Er-ping, M.D., Attending physician, Three Gorges University People’s Hospital, Yichang First People’s Hospital, Yichang 443000, Hubei Province, China
  • Supported by:

    the National Science Foundation for Young Scientists of China, No. 61308110

摘要:

文章快速阅读:


文题释义:
低密度脂蛋白受体相关蛋白:能够与多种结构及功能各异的配体相互作用,不仅可以对血脂的动态平衡及纤溶功能的稳定进行调节,而且能参与多种生长因子、细胞激酶生物学效应的发挥。
慢病毒:慢病毒属是反转录病毒科下的一个属,包括8种能够感染人和脊椎动物的病毒,原发感染的细胞以淋巴细胞和巨噬细胞为主,感染个体最终发病,显著特点是感染个体在出现典型的临床症状之前,大多经历长达数年的潜伏期之后缓慢发病,故被称为慢病毒。
摘要
背景:
最近研究表明低密度脂蛋白受体相关蛋白除了参与脂质代谢外,还参与调节炎症反应。
目的:观察慢病毒介导的低密度脂蛋白受体相关蛋白1-shRNA对大鼠骨关节炎模型软骨破坏的影响,以及对软骨组织中基质金属蛋白酶13表达的作用,初步判断低密度脂蛋白受体相关蛋白1对体内实验骨关节炎发病进程中的作用。
方法:SD大鼠64只分为4组,每组16只。①对照组大鼠未做手术为阴性对照组。②假手术组大鼠显露关节腔后及关闭切口,不切断交叉韧带和半月板。③模型+shLRP1组大鼠切断前交叉韧带切断加内侧半月板部分切除制作骨关节炎模型,造模后2 d用慢病毒介导的siRNA关节腔注射,每周1次,连续2周。④骨关节炎模型组,造模后关节腔注射阴性对照慢病毒。4组大鼠造模后5 d开始在自制的电动跑步机中跑步,每日跑步30 min,里程为500 m。造模2,4及6周处死各小组大鼠,观察软骨破坏程度和软骨组织中基质金属蛋白酶13表达。
结果与结论:①大体和病理切片观察发现:行关节腔注射慢病毒介导的siRNA抑制关节软骨的低密度脂蛋白受体相关蛋白1表达后,大鼠骨关节炎模型中软骨破坏加重,加速骨关节炎病程。②6周时:模型+shLRP1组Mankin’s评分明显高于其他3组(P < 0.05),模型+shLRP1组软骨细胞基质金属蛋白酶13阳性率明显高于其他3组(P < 0.05)。③结果说明:前叉韧带切断加内侧半月板部分切除并配合跑步机中慢跑锻炼能再现骨关节炎模型。慢病毒介导siRNA关节腔注射可干扰抑制关节软骨中的低密度脂蛋白受体相关蛋白1表达,对关节软骨有加重损伤的作用。

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程
ORCID: 0000-0002-8538-3658(杨二平)

关键词: 组织构建, 软骨组织工程, 骨关节炎, 低密度脂蛋白受体相关蛋白1, 慢病毒;前交叉韧带, 关节腔注射, 国家自然科学基金

Abstract:

BACKGROUND: Emerging evidence demonstrates that low density lipoprotein receptor-related protein 1 (LRP1) is involved in lipid metabolism and regulation of inflammatory reaction.
OBJECTIVE: To explore the effect of lentivirus-induced knockdown of low density lipoprotein receptor-related protein 1 on cartilage damage and matrix metalloproteinase 13 in a rat model of osteoarthritis, so as to assess the role of low density lipoprotein receptor-related protein 1 in the pathogenesis of osteoarthritis.
METHODS: Sixty-four Sprague-Dawley rats were included and ramdomly divided into four groups (n=16 for each): negative control group, no surgery; sham-surgery group, only the articular cavity of the knee was exposed; osteoarthritis plus shLRP1 group, rat osteoarthritis models were established by cutting anterior cruciate ligament and removing the medial meniscus partly followed by an intra-articular injection of lentivirus-mediated siRNA at 2 days after surgery, once a week for 2 consecutive weeks; osteoarthritis group, an intra-articular injection of the negative control lentivirus was performed after surgery. Rats in the four groups started running on the self-made electric treadmill from 5 days after modeling, 30 minutes per day, totally 500 meters. Cartilage damage and matrix metalloproteinase 13 expression in cartilage tissues were determined at 2, 4, 6 weeks after surgery.
RESULTS AND CONCLUSION: Gross and pathological observations showed that lentivirus-induced knockdown of low density lipoprotein receptor-related protein 1 aggravated cartilage damage in the rat model of osteoarthritis. At 6 weeks after surgery, Mankin’s score and matrix metalloproteinase 13 expression in the cartilage tissues in osteoarthritis plus shLRP1 group were significantly increased compared with other three groups (P < 0.05). These findings indicate that a simulation model of osteoarthritis is developed by cutting anterior cruciate ligament and removing the medial meniscus partly combined with running on the treadmill. Lentivirus-induced knockdown of LRP1 aggravates cartilage damage in a rat model of osteoarthritis

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

Key words: Osteoarthritis, Lentivirus, Anterior Cruciate Ligament, Tissue Engineering

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