中国组织工程研究

中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (46): 6868-6877.doi: 10.3969/j.issn.2095-4344.2016.46.004

• 软骨组织构建 cartilage tissue construction • 上一篇    下一篇

龙鳖胶囊对骨关节炎软骨细胞p38MAPK信号通路及NF-κB p65的影响

潘建科1，曹学伟1，刘  军1，谢  辉1，杨伟毅1，张  娴2   

  1. 广州中医药大学第二附属医院(广东省中医院)，1骨科，2中心实验室，广东省广州市  510120
  • 出版日期:2016-11-11 发布日期:2016-11-11
  • 通讯作者: 刘军，主任医师，教授，博士生导师，博士后合作导师，广州中医药大学第二附属医院(广东省中医院)骨科，广东省广州市 510120
  • 作者简介:潘建科，男，1988年生，广东省兴宁市人，汉族，2016年广州中医药大学毕业，博士，主要从事骨与关节退变及损伤的基础、临床和循证医学研究。
  • 基金资助:

    国家自然科学基金项目(81473698，81273781)；教育部高等学校博士点科研基金项目(20124425110004)；广东省科技计划项目 (2011B031700027)；广东省财政厅项目( [2014]157号)；广东省中医药管理局项目(20164020)；广东省中医院中医药科学技术研究专项(YK2013B2N19，YN2015MS15)

Effect of Longbie Capsule on p38MAPK pathway and nuclear factor-kappa B p65 in chondrocytes of osteoarthritis

Pan Jian-ke1, Cao Xue-Wei1, Liu Jun1, Xie Hui1, Yang Wei-yi1, Zhang Xian2   

  1. 1Department of Orthopedics, 2Central Laboratory, Second Affiliated Hospital, Guangzhou University of Chinese Medicine & Guangdong Provincial Hospital of Chinese Medicine, Guangzhou 510120, Guangdong Province, China
  • Online:2016-11-11 Published:2016-11-11
  • Contact: Liu Jun, Chief physician, professor, Doctoral supervisor, Postdoctoral co-supervisor, Department of Orthopedics, Second Affiliated Hospital, Guangzhou University of Chinese Medicine & Guangdong Provincial Hospital of Chinese Medicine, Guangzhou 510120, Guangdong Province, China
  • About author:Pan Jian-ke, M.D., Department of Orthopedics, Second Affiliated Hospital, Guangzhou University of Chinese Medicine & Guangdong Provincial Hospital of Chinese Medicine, Guangzhou 510120, Guangdong Province, China
  • Supported by:

    the National Natural Science Foundation of China, No. 81473698, 81273781; the Research Fund for the Doctoral Program of Higher Education of Chinese Ministry of Education, No. 20124425110004; the Science and Technology Program of Guangdong Province, No. 2011B031700027; the Project of Department of Finance of Guangdong Province, No. [2014]157; the Project of Traditional Chinese Medicine Bureau of Guangdong Province, No. 20164020; the Traditional Chinese Medicine Science and Technology Research Project of Guangdong Provincial Hospital of Chinese Medicine, No. YK2013B2N19, YN2015MS15

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摘要：

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文题释义：
p38MAPK信号通路：参与了细胞的生长发育及细胞间功能同步等多种生理过程，并与炎症、应激反应的调控密切相关，被认为是细胞信息传递的交汇点和共同通路。细胞外的多种应激原，如紫外线照射、细胞外高渗透压、放射线、炎性细胞因子(如白细胞介素1、肿瘤坏死因子α)、生长因子、热休克因子、细菌病原体及其产物等均可将p38激活，通过高度保守且较复杂的3级激酶“级联”程序转导信号。
NF-κB信号通路：在软骨细胞的促炎应激反应中发挥着核心作用，被认为是骨关节炎治疗中有吸引力的治疗靶点。NF-κB蛋白是一类广泛表达的转录因子，在细胞凋亡、细胞衰老、炎症炎症、免疫调节及肿瘤发生中起重要作用，NF-κB蛋白可被促炎细胞因子、趋化因子、应激因子和细胞外基质的降解产物所激活，活化的NF-κB分子触发一系列基因的表达，引起关节软骨破坏，导致骨关节炎的发病和进展。

摘要
背景：前期研究发现，龙鳖胶囊能消除膝骨关节炎大鼠膝关节肿胀，降低血清炎性细胞因子白细胞介素1β、白细胞介素6水平和升高抑炎细胞因子白细胞介素10水平。
目的：观察龙鳖胶囊对膝骨关节炎软骨细胞p38MAPK信号通路及NF-κB p65的影响。
方法：将60只SD大鼠随机分为5组，每组12只，模型组、低剂量组、中剂量组、高剂量组建立右侧膝骨关节炎大鼠模型，造模后第3天，低剂量组、中剂量组、高剂量组分别灌胃给予0.312 5，0.625，0.937 5 g/kg龙鳖胶囊，2次/d，连续4周；对照组不造模。灌胃2，4周后，取膝关节制成组织切片，免疫组织化学SP法检测软骨组织中MEK-3/6、p38、ATF2、NF-κB p65及其磷酸化产物P- MEK-3/6、P-p38、P-ATF2、P-NF-κB p65的表达。
结果与结论：给药2，4周后，各组细胞均有MEK-3/6、p38、ATF2、NF-κB p65及其磷酸化产物P- MEK-3/6、P-p38、P-ATF2、P-NF-κB p65阳性表达，模型组MEK-3/6、p38、ATF2、NF-κB p65及其磷酸化产物 P-MEK-3/6、P-p38、P-ATF2、P-NF-κB p65阳性细胞百分比均高于对照组(P < 0.01)，低、中、高剂量组上述指标阳性细胞百分比均低于模型组(P < 0.05)。结果表明，龙鳖胶囊可抑制MEK-3/6、p38、ATF2、NF-κB p65及其磷酸化产物P- MEK-3/6、P-p38、P-ATF2、P-NF-κB p65的过表达，抑制骨关节炎软骨细胞中p38MAPK和NF-κB信号通路活化，发挥减改善软骨退变、保护软骨的作用。

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: 0000-0002-7610-6667(刘军)

关键词: 组织构建, 软骨细胞, 龙鳖胶囊, 骨关节炎, 补肾活血, p38MAPK信号通路, NF-κB p65, 国家自然科学基金

Abstract:

BACKGROUND: Previous studies found that Longbie Capsule can relieve knee joint swelling of rats with knee osteoarthritis, and decrease the serum levels of interleukin-1β, interleukin-6 and interleukin-10.
OBJECTIVE: To observe the influence of Longbie Capsule on p38MAPK pathway and nuclear factor-κB p65 (NF-κB p65) in chondrocytes of osteoarthritis.
METHODS: Sixty Sprague-Dawley rats were equivalently randomized into five groups, and the rat models of right knee osteoarthritis were established in model, low-dose, middle-dose and high-dose groups. Three days later, rats in the latter three groups were respectively treated with 0.312 5, 0.625, and 0.937 5 g/kg Longbie Capsule via gastric lavage, twice daily for 4 consecutive weeks; while those in the control group received no intervention. At 2 and 4 weeks after treatment, the rat right knee joint was removed and made into paraffin tissue sections, and the expressions of MEK-3/6, p38, activating transcription factor 2 (ATF2), NF-κBp65, and their phosphorylated products of P-MEK-3/6, P-p38, P-ATF2, P-NF-κB p65 in cartilage tissue were detected by immunohistochemistry streptavidin-perosidase method.
RESULTS AND CONCLUSION: After 2 and 4 weeks of treatment, chondrocytes in each group could express MEK-3/6, p38, ATF2, NF-κB p65, P-MEK-3/6, P-p38, P-ATF2 and P-NF-κB p65, and the percentage of positive cells in the model group was significantly higher than that in the control group (P < 0.01), and the treatment groups (P < 0.05). These results indicate that Longbie Capsule cannot only inhibit the over expressions of MEK-3/6, p38, ATF2, NF-κB p65 and their phosphorylated products, but also inhibit the activation of p38MAPKs and NF-κB signaling pathway in chondrocytes of osteoarthritis, which is beneficial for cartilage degeneration and protecting the cartilage.

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: Drugs, Chinese Herbal, Osteoarthritis, Tissue Engineering

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