文献类型：期刊文献

中文题名：氨基葡萄糖和骨碎补联用对大鼠慢性骨关节炎的干预作用及机制

英文题名：Preventive Effect and Mechanisms of Glucosamine Combined with Rhizoma Drynariae on Chronic Osteoarthritis Rats

作者：周艳丽[1];陈世杰[2];劳文艳[1,3];朱杨雄[1];赵晓红[1,3]

第一作者：周艳丽

通讯作者：Zhao, Xiaohong

机构：[1]北京联合大学功能食品科学技术研究院;[2]北京赛升药业股份有限公司;[3]北京联合大学生物活性物质与功能食品北京市重点实验室

第一机构：北京联合大学应用文理学院|北京联合大学生物化学工程学院功能食品科学技术研究院

年份：2018

卷号：39

期号：3

起止页码：213-221

中文期刊名：食品科学

外文期刊名：Food Science

收录：CSTPCD;;EI(收录号：20181705054221);Scopus;北大核心:【北大核心2017】；CSCD:【CSCD_E2017_2018】；

基金：北京联合大学生物活性物质与功能食品北京市重点实验室开放课题(Zk70201501);2017年"启明星"大学生科技创新项目(01711417XJ073)

语种：中文

中文关键词：氨基葡萄糖;骨碎补;骨关节炎;关节软骨

外文关键词：glucosamine; drynaria rhizome; osteoarthritis; articular cartilage

摘要：目的:探讨氨基葡萄糖(glucosamine,GLU)和骨碎补(drynaria rhizome,DR)联合用药对大鼠慢性骨关节炎(osteoarthritis,OA)的干预作用及机制。方法:关节腔注射白陶土-鹿角菜胶诱发大鼠急性骨关节损伤,结合跑步运动,建立大鼠慢性OA模型。造模前开始灌胃,分别为750 mg/kg GLU(GLU单独作用组)、150 mg/kg DR(DR单独作用组)、125 mg/kg GLU+25 mg/kg DR(低剂量联合组)、250 mg/kg GLU+50 mg/kg DR(中剂量联合组)、750 mg/kg GLU+150 mg/kg DR(高剂量联合组)、无菌水(空白组、模型组)和2 mg/kg双氯芬酸钠(diclofenac sodium,DS;阳性对照组),直至6周跑台运动后结束。测量大鼠骨关节肿胀度;取膝关节制作石蜡组织切片,分别进行苏木精-伊红和番红-O-固绿染色,观察关节软骨组织学病理变化;用酶联免疫吸附检测(enzyme-linked immunosorbent assay,ELISA)法测定血清金属基质蛋白酶-3(matrix metalloproteinase-3,MMP-3)、金属基质蛋白酶抑制剂(tissue inhibitor of metalloproteinase-1,T I M P-1)、诱导型N O合成酶(inducible nitric oxide synthase,i N O S)、白介素-1β(i n t e r l e u k i n-1β,IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)水平,黄嘌呤氧化酶法测定血清中超氧化物歧化酶(superoxide dismutase,SOD)活力,硝酸还原酶法测定血清和滑膜组织中NO浓度,免疫组化检测关节软骨Ⅱ型胶原的表达情况。结果:与空白组相比,模型组大鼠关节肿胀,软骨破坏;血清中MMP-3、TIMP-1质量浓度和MMP-3/TIMP-1质量浓度比值显著升高(P<0.05);i NOS活力、IL-1β、NO和TNF-α质量浓度显著增加(P<0.05);SOD活力和关节软骨Ⅱ型胶原含量显著降低(P<0.05)。与模型组相比,GLU和DR单独作用及中、高剂量联合作用均能减轻关节肿胀,改善膝关节组织病理学异常现象;明显抑制血清MMP-3、TIMP-1、NO、IL-1β、TNF-α质量浓度和i NOS活力的增加(P<0.05);抑制SOD活力的降低(P<0.05);抑制关节软骨Ⅱ型胶原含量的降低(P<0.05)。其中,高剂量联合组效果最好,然后依次是GLU、DR、中剂量联合组、低剂量联合组。结论:GLU和DR单独和联合作用都能够减轻骨关节损伤,联合作用需要达到有效作用剂量才有明显的抑制作用。其作用机制与调节MMP-3/TIMP-1平衡,增加SOD活力和Ⅱ型胶原含量,降低i NOS活力和IL-1β、TNF-α、NO水平有关。这些研究为预防和治疗OA的功能性食品的研发提供了理论依据。
Objective： To investigate the preventive effect and mechanisms of glucosamine（GLU） combined with drynariae rhizome（DR） on osteoarthritis（OA） of rats. Methods： A rat model of chronic arthritis was established by injection of kaolin and carrageen-λ into the articular cavity to induce acute osteoarticular injury combined with run, which was preceded by intragastric administration of GLU（750 mg/kg）, DR（150 mg/kg）, low-, medium-and high-dose GLU ＋ DR（125 mg/kg GLU ＋ 25 mg/kg DR, 250 mg/kg GLU ＋ 50 mg/kg DR, and 750 mg/kg GLU ＋ 150 mg/kg DR）, diclofenac sodium（DS, 2 mg/kg, positive control）, and sterile distilled water（10 m L/kg, for control and model groups） until the end of a 6 week run on a threadmill. Arthroncus in rats was measured. Tissue sections of knee joints were prepared by paraffin embedding and staining with hematoxylin eosin and saffron-O-solid green to observe the pathological changes of articular cartilage. The levels of matrix metalloproteinase-3（MMP-3）, tissue inhibitor of metalloproteinase-1（TIMP-1）, inducible nitric oxide synthase（i NOS）, interleukin-1β（IL-1β）, and tumor necrosis factor-α（TNF-α） in serum were detected with enzyme-linked immunosorbent assay（ELISA） kits. Superoxide dismutase（SOD） activity and nitric oxide（NO） content in serum were measured by xanthine oxidase and nitrate reductase methods, respectively. The expression of type Ⅱ collagen was detected by immunohistochemistry. Results： Compared with the control group, arthroncus in rats in the model group was increased and the cartilage was destroyed. Moreover, MMP-3 and TIMP-1 concentrations and MMP-3/TIMP-1 ratio were significantly increased as well as i NOS activity and of IL-1β, NO and TNF-α concentrations（P 0.05）, while SOD activity and type Ⅱcollagen expression were significantly declined（P 0.05）. Compared with the model group, GLU and DR alone as well as their combination at medium and high doses could reduce arthroncus, improve pathological abnormality in knee joint tissue, significantly inhibit the increase in MMP-3, TIMP-1, IL-1β, NO and TNF-α contents and i NOS activity, and significantly suppress the decrease in SOD activity and type II collagen content（P 0.05）, and the best effect was observed with highdose GLU ＋ DR followed by GLU, DR, and GLU ＋ DR at medium and low doses. Conclusion： Separate and combined GLU and RD can attenuate osteoarticular injury, but an effective dose is required for the combined treatment. The potential mechanism of action may be related to the regulation of MMP-3/TIMP-1 balance, resulting in increased SOD activity and type Ⅱ collagen content as well as decreased i NOS activity and IL-1β, TNF-α and NO contents. This study can provide a theoretical basis for the development of functional foods for the prevention and treatment of OA.