吉林大学学报(医学版) ›› 2023, Vol. 49 ›› Issue (3): 617-624.doi: 10.13481/j.1671-587X.20230309

• 基础研究 • 上一篇    下一篇

防风野生品对大鼠类风湿性关节炎的改善作用及其机制

林喆1,王泰栋1,黄晓巍1,马超1,庄雪峰1,王雨辰1,林贺1,郭骏骐1(),律广富2()   

  1. 1.长春中医药大学药学院中药药理学实验室,吉林 长春 130117
    2.长春中医药大学 吉林省人参 科学研究院中药药理组,吉林 长春 130117
  • 收稿日期:2022-07-08 出版日期:2023-05-28 发布日期:2023-06-20
  • 通讯作者: 郭骏骐,律广富 E-mail:kjcchgk@163.com;message219@163.com
  • 作者简介:林 喆(1966-),男,吉林省长春市人,教授,医学硕士,主要从事中药药理学方面的研究。
  • 基金资助:
    吉林省发改委产业技术研究与开发项目(2020C032-2);吉林省科技厅医药健康产业发展专项项目(20210401067YY);吉林省中医药管理局中医药科技项目(2022014)

Improvement effect of Saposhnikoviae Radix wild product on rheumatoid arthritis in rats and its mechanism

Zhe LIN1,Taidong WANG1,Xiaowei HUANG1,Chao MA1,Xuefeng ZHUANG1,Yuchen WANG1,He LIN1,Junqi GUO1(),Guangfu LYU2()   

  1. 1.Labratory of Pharmacology of Chinese Medicine,School of Pharmacy,Changchun University of Chinese Medicine,Changchun 130117,China
    2.Department of Pharmacology of Traditional Chinese Medicine,Jilin Ginseng Academy,Changchun University of Chinese Medicine,Changchun 130117,China
  • Received:2022-07-08 Online:2023-05-28 Published:2023-06-20
  • Contact: Junqi GUO,Guangfu LYU E-mail:kjcchgk@163.com;message219@163.com

摘要:

目的 探讨防风野生品对类风湿性关节炎的治疗作用和对核因子κB(NF-κB)信号通路的影响,为类风湿性关节炎的治疗提供依据。 方法 50只大鼠随机分为对照组、模型组、地塞米松组、低剂量防风组和高剂量防风组,每组10只。除对照组外,其余各组大鼠采用尾根部和左后足跖处皮内注射Ⅱ型胶原制备类风湿性关节炎模型,模型制备成功后,地塞米松组大鼠给予地塞米松(2 mg·kg-1),低和高剂量防风组大鼠给予防风野生品(0.45和0.90 g·kg-1),对照组和模型组大鼠给予等体积生理盐水。检测各组大鼠关节炎指数(AI)评分和足肿胀程度,计算各组大鼠脏器指数,采用酶联免疫吸附测定(ELISA)法检测各组大鼠血清中肿瘤坏死因子α(TNF-α)、白细胞介素1β(IL-1β)和白细胞介素6(IL-6)水平,HE染色观察各组大鼠滑膜组织病理形态表现,Western blotting法检测各组大鼠滑膜组织中NF-κB、NF-κB抑制因子α(IκB-α)、IκB-α激酶β(IKK-β)、环氧化酶2(COX-2)和TNF-α蛋白表达水平。 结果 与对照组比较,模型组大鼠AI评分明显升高(P<0.01),足肿胀程度明显加重(P<0.01),胸腺指数明显升高(P<0.01);与模型组比较,地塞米松组和低及高剂量防风组大鼠AI评分明显降低(P<0.01),足肿胀程度明显减轻(P<0.05或P<0.01),胸腺指数明显降低(P<0.01)。ELISA法检测,与对照组比较,模型组大鼠血清中TNF-α、IL-1β和IL-6水平明显升高(P<0.01);与模型组比较,地塞米松组、低和高剂量防风组大鼠血清中TNF-α、IL-1β和IL-6水平明显降低(P<0.01)。HE染色,对照组大鼠滑膜组织未见异常,模型组大鼠滑膜组织增生并伴有大量炎症细胞浸润;与模型组比较,地塞米松组和低及高剂量防风组大鼠滑膜组织增生和炎症细胞浸润程度减轻。Western blotting法检测,与对照组比较,模型组大鼠滑膜组织中NF-κB、IKK-β、COX-2和TNF-α蛋白表达水平明显升高(P<0.01),IκB-α蛋白表达水平明显降低(P<0.01);与模型组比较,地塞米松组、低和高剂量防风组大鼠滑膜组织中NF-κB、IKK-β、COX-2和TNF-α蛋白表达水平明显降低(P<0.01),IκB-α蛋白表达水平明显升高(P<0.01)。 结论 防风野生品能够通过降低机体促炎细胞因子水平和抑制滑膜组织中NF-κB信号通路的过度激活改善关节组织损伤,从而发挥对类风湿性关节炎的治疗作用。

关键词: 防风野生品, 类风湿性关节炎, 核因子κB, 地塞米松, 肿瘤坏死因子α

Abstract:

Objective To discuss the therapeutic effect of Saposhnikoviae Radix on rheumatoid arthritis and its effect on the nuclear factor-κB (NF-κB) signaling pathway,and to provide the basis for the treatment of rheumatoid arthritis. Methods Fifty rats were randomly divided into control group, model group, dexamethasone group, low dose of Saposhnikoviae Radix group, and high dose of Saposhnikoviae Radix group,and there were 10 rats in each group; except for control group, the rats in the other groups were given intradermal injection of type Ⅱ collagen at the root of the tail and left posterior foot to prepare the rheumatoid arthritis models; after the models were prepared successfully, the rats in dexamethasone group were given dexamethasone (2 mg·kg-1),the rats in low and high doses of Saposhnikoviae Radix groups were given Saposhnikoviae Radix wild product (0.45 and 0.90 g·kg-1), and equal volume of normal saline was given to the rats in control group and model group.The arthritis index(AI) and foot swelling degrees of the rats in various groups were recorded, the organ coefficients of the rats in various groups were calculated; enzyme-linked immunosorben assay (ELISA) method was used to detect the levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6) in serum of the rats in various groups, and the expression levels of NF-κB,NF-κB inhibitor α (IkB-α), IκB-α kinase β (IKK-β), cyclooxidase-2 (COX-2),and TNF-α proteins in synovial tissue of the rats in various groups were detected by Western blotting method. Results Compared with control group, the AI score and swelling degree of foot of the rats in model group were significantly decreased (P<0.01), while the thymus index was significantly increased (P<0.01); compared with model group, the AI score and swelling degree of foot of the rats in dexamethasone group and low and high doses of Saposhnikoviae Radix groups were significantly decreased (P<0.05 or P<0.01), and the thymus indexes were significantly decreased(P<0.01).The ELISA results showed that compared with control group,the levels of TNF-α, IL-1β,and IL-6 in serum of the rats in model group were increased(P<0.01); compared with model group,the levels of TNF-α, IL-1β,and IL-6 in serum of the rats in dexamethasone group, low and high doses of Saposhnikoviae Radix groups were significantly decreased (P<0.01).The HE staing results showed that the synovial tissue of the rats in control group was normal,and the synovial tissue of the rats in model group showed hyperplasia accompanied with extensive infiltration of inflammatory cells; compared with model group, the degrees of proliferation of synovial tissue and infiltration of inflammatory cells of the rats in dexamethasone group, low and high doses of Saposhnikoviae Radix groups were decreased.The Western blotting results showed that compared with control group, the expression levels of NF-κB,IKK-β,COX-2, and TNF-α proteins in synovial tissue of the rats in model group were increased (P<0.01),the expression level of IκB-α protein was decreased (P<0.01);compared with model group, the expression levels of NF-κB,IKK-β,COX-2, and TNF-α proteins in synovial tissue of the rats in dexamethasone group, low and high doses of Saposhnikoviae Radix groups were decreased (P<0.01),the expression level of IκB-α protein was increased (P<0.01). Conclusion Saposhnikoviae Radix wild product can improve the joint injury by reducing the levels of proinflammatory cytokines and inhibiting the excessive activation of NF-κB signaling pathway in synovial tissue, thus playing a therapeutic role in the rheumatoid arthritis.

Key words: Saposhnikoviae Radix wild product, Rheumatoid arthritis, Nuclear factor kappa-B, Dexamethasone, tumor necrosis factor-α

中图分类号: 

  • R965.1