右归丸经IL-6/STAT3信号通路对膝骨关节炎模型大鼠软骨组织退变的调控机制

颜春鲁; 安方玉; 刘永琦; 王继龙; 赵磊; 夏鹏飞; 周婷; 石国秀; 赵文坤; 杨晓蓉

doi:10.13422/j.cnki.syfjx.20192303

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右归丸经IL-6/STAT3信号通路对膝骨关节炎模型大鼠软骨组织退变的调控机制

经典名方 | 更新时间：2021-02-09

- 右归丸经IL-6/STAT3信号通路对膝骨关节炎模型大鼠软骨组织退变的调控机制
- Regulatory Mechanism of Youguiwan on Articular Cartilage Degeneration via IL-6/STAT3 Signaling Pathway in Rats with Knee Osteoarthritis
- 中国实验方剂学杂志 2020年26卷第1期页码：17-23
- 作者机构：
  
  甘肃中医药大学　甘肃省高校重大疾病分子医学与中医药防治研究重点实验室，甘肃省高校中(藏)药化学与质量研究省级重点实验室，敦煌医学与转化教育部重点实验室，兰州　 730000
- 作者简介：
  
  [第一作者]　颜春鲁，硕士，副教授，从事中药抗骨病的分子生物学机制研究，E-mail：yanchl1979@126.com
  *安方玉，硕士，副教授，从事中药抗骨病的分子生物学机制研究，E-mail：anfyuwsmh@163.com
- 基金信息：
  
  甘肃省高等学校科研项目(2018A-043);甘肃省中医药管理局科研项目(GZK-2017-2);敦煌医学与转化省部共建教育部重点实验室开放基金项目(DHYX17-10);甘肃省高校中（藏）药化学与质量研究省级重点实验室开放基金项目(zzy-2018-01);甘肃省高校重大疾病分子医学与中医药防治研究重点实验室开放基金项目(FZYX17-18-11)
- DOI：10.13422/j.cnki.syfjx.20192303
  中图分类号： R2-0; R22; R285.5; R289
- 收稿日期：2019-05-22，
  
  网络出版日期：2019-08-20，
  
  纸质出版日期：2020-01-05
- 稿件说明：
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颜春鲁, 安方玉, 刘永琦, 等. 右归丸经IL-6/STAT3信号通路对膝骨关节炎模型大鼠软骨组织退变的调控机制[J]. 中国实验方剂学杂志, 2020,26(1):17-23.

Chun-lu YAN, Fang-yu AN, Yong-qi LIU, et al. Regulatory Mechanism of Youguiwan on Articular Cartilage Degeneration via IL-6/STAT3 Signaling Pathway in Rats with Knee Osteoarthritis[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(1): 17-23.
颜春鲁, 安方玉, 刘永琦, 等. 右归丸经IL-6/STAT3信号通路对膝骨关节炎模型大鼠软骨组织退变的调控机制[J]. 中国实验方剂学杂志, 2020,26(1):17-23. DOI： 10.13422/j.cnki.syfjx.20192303.

Chun-lu YAN, Fang-yu AN, Yong-qi LIU, et al. Regulatory Mechanism of Youguiwan on Articular Cartilage Degeneration via IL-6/STAT3 Signaling Pathway in Rats with Knee Osteoarthritis[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(1): 17-23. DOI： 10.13422/j.cnki.syfjx.20192303.

摘要

目的：

观察右归丸对膝骨关节炎(KOA)模型鼠软骨组织信号转导和转录激活因子3(STAT3)和白细胞介素-6(IL-6)表达水平的影响。

方法：

将大鼠随机分为假手术组、模型组、硫酸氨基葡萄糖组、右归丸高、中、低剂量组，每组10只。采用改良Hulth法制备大鼠KOA模型，假手术组和模型组给予等体积生理盐水灌胃，右归丸高、中、低剂量组分别给予右归丸4.8，2.4，1.2 g·kg

-1

灌胃，硫酸氨基葡萄糖组给予硫酸氨基葡萄糖0.17 g·kg

-1

灌胃，连续给药8周。干预结束24 h后股动脉采血处死各组大鼠，取鼠膝关节软骨，采用苏木素-伊红(HE)染色法观察各组软骨的病理改变，并进行Mankin评分；免疫组化法检测各组关节软骨组织中STAT3，超氧化物歧化酶3(SOD3)和Wnt抑制因子1(WIF1)的表达；实时荧光定量聚合酶链式反应(Real-time PCR)检测软骨组织中IL-6 mRNA的表达；蛋白免疫印迹法(Western blot)检测各组关节软骨组中WIF1蛋白的表达。

结果：

与假手术组比较，模型组大鼠软骨组织Makin评分明显升高，软骨组织STAT3在蛋白水平上的表达明显增加和IL-6 mRNA水平上的表达显著增加，WIF1在蛋白水平上的表达显著降低(

0.01)；模型组关节软骨边缘严重破坏，软骨细胞排列紊乱。与模型组比较，右归丸高剂量干预组大鼠软骨组织Makin评分，STAT3的在蛋白水平上的表达明显降低，右归丸各干预组IL-6 mRNA水平上的表达显著降低，WIF1在蛋白水平上的表达显著增加(

0.05，

0.01)，软骨结构趋于正常，软骨细胞分布仅偶见不均，关节软骨表面欠光滑。

结论：

右归丸能显著改善KOA大鼠的关节软骨退变，抑制KOA中软骨组织的炎症反应，这可能与其抑制STAT3和IL-6的表达有关。

Abstract

Objective:

To observe the effect of Youguiwan on the levels of cartilage transducers and activators of transcription 3 (STAT3) and interleukin-6 (IL-6) in rats with knee osteoarthritis (KOA).

Method:

Sixty SD rats were randomly divided into six groups: sham control group

model group

glucosamine sulfate group and Youguiwan (high

middle and low-dose) groups. The modified Hulth method was used to prepare KOA models for 6 weeks. The shame control group and the model group were treated with normal saline

Youguiwan high

middle

low-dose groups received Youguiwan 4.8

2.4

1.2 g·kg

-1

by gavage respectively

and the glucosamine sulfate group was treated with glucosamine sulfate 0.17 g·kg

-1

. The rats were administrated for 8 weeks according to the dose. After intervention

each group was put to death by femoral artery blood collection

and the keen cartilages of the rats were collected. The pathological changes were observed by htoxylin eosin (HE) staining method

and Mankin score was evaluated. The expressions of STAT3

superoxide dismutase3(SOD3) and Wnt inhibitory factor 1(WIF1) in articular cartilage were detected by immunohistochemistry. The expressions of IL-6 mRNA in articular cartilage were detected by quantitative real-time fluorescence polymerase chain reaction (Real-time PCR). The expression of WIF1 in articular cartilage was detected by Western blot.

Result:

Compared with the sham control group

the Makin score was obviously increased in the model group

the protein expression of STAT3 was increased significantly

the mRNA of IL-6 was raised significantly

but the protein expression of WIF1 was decreased significantly (

0.01)

articular cartilage was seriously damaged

and chondrocytes were arranged in disorder. Compared with the model group

the Makin score was declined obviously in the high-dose Youguiwan group

the protein expression of STAT3 was significantly reduced

the mRNA expression of IL-6 was significantly reduced in Youguiwan treatment group

while the protein expression of WIF1 was significantly increased (

0.05

0.01)

the cartilage structure returned to be normal

the chondrocytes distribution was uneven

and articular cartilage surface was not smooth.

Conclusion:

Youguiwan could significantly improve the articular cartilage degeneration of KOA rats

and inhibit the inflammation of chondrocytes

which may be related to the suppression of STAT3 and IL-6 expression.

关键词

Keywords

references

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