中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (27): 4375-4380.doi: 10.3969/j.issn.2095-4344.1388

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

补肾活血汤干预骨质疏松症并骨关节炎模型兔软骨细胞NF-KBp65蛋白的表达

张  波1,张开伟2,马文娟3,沈冯君2,陈海霞1   

  1. (1贵州中医药大学,贵州省贵阳市  550000;2贵阳中医学院第一附属医院,贵州省贵阳市  550000;3贵州省金沙县中医院,贵州省毕节市  551800)
  • 收稿日期:2019-03-28 出版日期:2019-09-28 发布日期:2019-09-28
  • 通讯作者: 张开伟,博士,主任医师,贵阳中医学院第一附属医院,贵州省贵阳市 550001
  • 作者简介:张波,男,1990年生,贵州省德江县人,土家族,贵州中医药大学在读硕士,主要从事骨与关节病变研究。
  • 基金资助:

    贵州省科技厅联合基金(黔科合中药字[2011]LKZ7022号),项目负责人:张开伟;贵州省中管局科研项目(QZYY2013-17号),项目负责人:张开伟

Effect of Bushen Huoxue Decoction on the expression level of nuclear factor-KBp65 protein in chondrocytes of rabbit models of osteoporosis and osteoarthritis 

Zhang Bo1, Zhang Kaiwei2, Ma Wenjuan3, Shen Fengjun2, Chen Haixia1   

  1.  (1Guiyang University of Traditional Chinese Medicine, Guiyang 550000, Guizhou Province, China; 2the First Affiliated Hospital of Guiyang University of Traditional Chinese Medicine, Guiyang 550000, Guizhou Province, China; 3Jinsha County Hospital of Traditional Chinese Medicine of Guizhou Province, Bijie 551800, Guizhou Province, China)
  • Received:2019-03-28 Online:2019-09-28 Published:2019-09-28
  • Contact: Zhang Kaiwei, MD, Chief physician, the First Affiliated Hospital of Guiyang University of Traditional Chinese Medicine, Guiyang 550000, Guizhou Province, China
  • About author:Zhang Bo, Master candidate, Guiyang University of Traditional Chinese Medicine, Guiyang 550000, Guizhou Province, China
  • Supported by:

    the Combined Foundation of Guizhou Provincial Department of Science and Technology, No. [2011]LKZ7022 (to ZKW); the Research Project of Guizhou Provincial Administration of Traditional Chinese Medicine, No. QZYY2013-17 (to ZKW)

摘要:

文章快速阅读:

文题释义:

信号通路(NF-KB):广泛存在于细胞基质内的一种核转录因子,在未受刺激的细胞中,Kappa B 抑制蛋白结合在NF-KB的核定位序列上,从而将NF-KB阻滞在细胞质,使其不能进入细胞核发挥转录调控作用,但受到各种因子如白细胞介素1、白细胞介素6、肿瘤坏死因子α等刺激后,Kappa B抑制蛋白被降解,NF-KB被激活后F转移到细胞核内结合到其靶基因的启动子上,通过启动靶基因的转录,进而调控细胞的生长和炎症反应。
阿仑膦酸钠:适用于治疗绝经后妇女的骨质疏松症,以预防髋部和脊柱骨折(椎骨压缩性骨折),也适用于男性骨质疏松症以增加骨量。动物研究发现,在细胞水平,阿仑膦酸钠对骨吸收部位特别是破骨细胞作用的部位有亲嗜性。正常情况下,破骨细胞黏附于骨表面但并不粗糙,而粗糙的边缘则是骨吸收活跃的标志。阿仑膦酸钠不影响破骨细胞的聚集或黏附,但它确实能够抑制破骨细胞的活性。
摘要
背景:
临床研究发现肾虚血淤是骨性关节炎、骨质疏松症共同病因、病机,运用补肾活血法治疗该类病变疗效肯定。
目的:进一步验证补肾活血汤对兔软骨细胞NF-KBp65蛋白表达的影响。
方法:雌性新西兰大白兔37只,由贵阳医学院动物实验中心提供,实验方案经贵阳医学院动物实验伦理委员会批准(批准号:2015-024)。随机选取30只大白兔编号后随机均分为3组,分别建立骨质疏松模型、骨关节炎模型、骨质疏松+骨关节炎模型。剩余7只用于制备7种含药血清(分别为正常组、低剂量补肾活血汤组、中剂量补肾活血汤组、高剂量补肾活血汤组、盐酸氨基葡萄糖组、阿仑膦酸钠组、盐酸氨基葡萄糖+阿仑磷酸钠组)。采用Western-blot检测7组药物对3组模型兔软骨细胞NF-KBp65蛋白表达的影响。
结果与结论:①中、高剂量组有下调细胞中NF-KBp65蛋白的作用,与正常组对比差异有显著性(P < 0.05),其中以高剂量组作用最为明显,与低、中剂量组对比差异有显著性意义(P < 0.05);②高剂量组下调NF-KBp65蛋白表达的作用较阿仑膦酸钠组和盐酸氨基葡萄糖组明显(P < 0.05);③高剂量组与盐酸氨基葡萄糖+阿仑膦酸钠组在骨质疏松症组对比差异有显著性意义(t=7.597,P < 0.05),但在骨关节炎组及骨关节炎+骨质疏松症组中则差异无显著性意义(P > 0.05),说明高剂量组下调NF-KBp65蛋白表达的作用优于阿仑膦酸钠组及盐酸氨基葡萄糖组,与盐酸氨基葡萄糖+阿仑膦酸钠组大体相当;④结果表明,补肾活血汤能下调NF-KBp65蛋白表达,从而减轻或抑制骨质疏松症和骨关节炎的炎症反应,间接或直接起到保护或修复软骨细胞的作用。

关键词: 骨质疏松症, 骨关节炎, 蛋白质印迹法, 补肾活血汤, NF-KBp65

Abstract:

BACKGROUND: Kidney deficiency and blood stasis are the common etiology and pathogenesis of osteoarthritis and osteoporosis. The method of nourishing kidney and activating blood circulation is effective for treating these diseases.
OBJECTIVE: To further investigate the effects of Bushen Huoxue Decoction on the expression level of nuclear factor-KBp65 protein in rabbit chondrocytes.
METHODS: Thirty-seven female New Zealand white rabbits were provided by the Laboratory Animal Center of Guiyang Medical University, and the study was approved by the Experimental Animal Ethics Committee of Guiyang Medical University, approval number: 2015-024. Thirty rabbits were randomly divided into three groups, and the osteoporosis, osteoarthritis and osteoporosis + osteoarthritis models, were separately established. The remaining seven rabbits were used to prepare the serum containing drug, such as normal, low-, moderate-, and high-dose Bushen Huoxue Decoction, glucosamine hydrochloride, alendronate sodium, and glucosamine hydrochloride + alendronate sodium groups. The effect of seven groups of drugs on the expression level of nuclear factor-KBp65 protein in rabbit chondrocytes of three kinds of models was detected by western blot assay.
RESULTS AND CONCLUSION: (1) Moderate- and high-dose Bushen Huoxue Decoction down-regulated nuclear factor-KBp65 protein level in cells, which was significantly different from the normal group (P < 0.05). The effect was most obvious in the high-dose Bushen Huoxue Decoction group, compared with the low- and the moderate-dose groups (P < 0.05). (2) The down-regulation effect of unclear factor-KBp65 protein expression in the high-dose Bushen Huoxue Decoction group was significantly higher than that in the glucosamine hydrochloride and alendronate sodium groups (P < 0.05). (3) There was a significant difference between the high-dose Bushen Huoxue Decoction and glucosamine hydrochloride + alendronate sodium groups in the osteoporosis group (t=7.597, P < 0.05), but there was no significant difference between the osteoarthritis and osteoporosis + osteoarthritis groups (P > 0.05). Therefore, the down-regulation effect of unclear factor-KBp65 protein expression in the high-dose Bushen Huoxue Decoction group was better than that in the glucosamine hydrochloride and alendronate sodium groups, which was similar to that in the glucosamine hydrochloride + alendronate sodium group. (4) These results indicate that Bushen Huoxue Decoction can down-regulate the expression of nuclear factor-KBp65 protein, so as to reduce or inhibit the inflammatory reaction of osteoporosis and osteoarthritis, and indirectly or directly plays a role in protecting or repairing chondrocytes.

Key words: osteoporosis, osteoarthritis, western blot assay, Bushen Huoxue Decoction, nuclear factor-KBp65

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