Mechanism of Zhiwang Decoction Combined with Methotrexate Against Bone Destruction Through Regulating RANKL/OPG Pathway in Rats with Collagen-induced Arthritis

Ru MA; Zi-jia GUO; Qing-wen TAO; Ze-ran YAN; Zhen-quan LIU; Yuan XU

您当前的位置：

首页 >

文章列表页 >

Mechanism of Zhiwang Decoction Combined with Methotrexate Against Bone Destruction Through Regulating RANKL/OPG Pathway in Rats with Collagen-induced Arthritis

更新时间：2022-02-07

- Mechanism of Zhiwang Decoction Combined with Methotrexate Against Bone Destruction Through Regulating RANKL/OPG Pathway in Rats with Collagen-induced Arthritis
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 28, Issue 5, Pages: 46-54(2022)
- 作者机构：
  
  1.西安市公共卫生中心（西安市应急医疗中心），西安 710200
  2.内蒙古医科大学中医学院，呼和浩特 010110
  3.中日友好医院中医风湿病科，免疫炎性疾病北京市重点实验室，北京 100029
  4.北京中医药大学国家体制与治未病研究院，北京 100029
- 作者简介：
- 基金信息：
- DOI：
  CLC： R22;R242;R285.5;R2-031;R593.22;R255.6
- Received：17 August 2021，
  
  Published Online：14 January 2022，
  
  Published：05 March 2022
- 稿件说明：
移动端阅览
马茹,郭子嘉,陶庆文等.治尪汤联合甲氨蝶呤调节RANKL/OPG通路改善胶原诱导关节炎大鼠骨破坏的作用机制[J].中国实验方剂学杂志,2022,28(05):46-54.

MA Ru,GUO Zi-jia,TAO Qing-wen,et al.Mechanism of Zhiwang Decoction Combined with Methotrexate Against Bone Destruction Through Regulating RANKL/OPG Pathway in Rats with Collagen-induced Arthritis[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(05):46-54.
马茹,郭子嘉,陶庆文等.治尪汤联合甲氨蝶呤调节RANKL/OPG通路改善胶原诱导关节炎大鼠骨破坏的作用机制[J].中国实验方剂学杂志,2022,28(05):46-54. DOI：

MA Ru,GUO Zi-jia,TAO Qing-wen,et al.Mechanism of Zhiwang Decoction Combined with Methotrexate Against Bone Destruction Through Regulating RANKL/OPG Pathway in Rats with Collagen-induced Arthritis[J].Chinese Journal of Experimental Traditional Medical Formulae,2022,28(05):46-54. DOI：

摘要

目的

探究治尪汤联合甲氨蝶呤改善胶原诱导关节炎模型（CIA）大鼠的骨破坏作用及机制。

方法

SD雄性大鼠随机分为正常组，模型组，甲氨蝶呤组（1.5 mg·kg

-1

），甲氨蝶呤+治尪汤低（40.8 g·kg

-1

），高（102.0 g·kg

-1

）剂量组。除正常组外，其余大鼠分别于第1，7天尾部注射浓度为2 g·L

-1

的Ⅱ型胶原乳剂50 μL建立CIA模型；第8天开始连续给药21 d，每7 d检测1次体质量、足趾厚度，进行关节炎指数（AI）评分，酶联免疫吸附测定法检测血清肿瘤坏死因子-

（TNF-

），白细胞介素-6（IL-6），白细胞介素-1

（IL-1

）含量，抗酒石酸酸性磷酸酶染色法检测踝关节破骨细胞的数量，免疫组化（IHC），实时荧光定量聚合酶链式反应（Real-time PCR），蛋白免疫印迹法（Western blot）检测踝关节核转录因子-

B受体活化因子配体（RANKL），骨保护素（OPG）的表达。

结果

与正常组比较，模型组大鼠体质量显著降低（

0.01），足趾厚度显著增加

（P

0.01），踝关节AI评分显著增加

（P

0.01），血清TNF-

，IL-6，IL-1

含量显著升高（

0.01），踝关节破骨细胞数量显著增加（

0.01），踝关节RANKL mRNA和蛋白表达显著升高（

0.01），OPG mRNA和蛋白表达明显降低（

0.05），RANKL/OPG显著升高（

0.01）。与模型组比较，甲氨蝶呤+治尪汤低、高剂量可改善大鼠体质量，降低足趾厚度，AI评分，RANKL mRNA和蛋白表达并增加OPG mRNA和蛋白表达，使RANKL/OPG降低（

0.05，

0.01），且甲氨蝶呤+治尪汤高剂量效果最好，甲氨蝶呤+治尪汤低剂量次之，甲氨蝶呤再次之。

结论

治尪汤联合甲氨蝶呤能更好地改善类风湿关节炎的骨破坏，可能是通过抑制RANKL/OPG通路实现的，本研究为中西医结合方案治疗类风湿关节炎骨破坏提供了实验基础。

Abstract

Objective

To explore the effect and mechanism of Zhiwang decoction combined with methotrexate （MTX） against bone destruction in rats with collagen-induced arthritis （CIA）.

Method

Male SD rats were randomly divided into a normal group， a model group， an MTX group （1.5 mg·kg

-1

）， and low-（40.8 g·kg

-1

） and high-（102.0 g·kg

-1

）combination groups （MTX+ Zhiwang decoction）. The CIA model was induced in rats except for those in the normal group by tail injection of 50 μL of 2 g·L

-1

type Ⅱ collagen on the 1st and 7th days. From the 8th day， the rats were treated correspondingly for 21 days. The body weight， toe thickness， and arthritis index（AI）were measured every seven days. Enzyme-linked immunosorbent assay （ELISA） was used to detect the content of inflammatory factors in the serum， such as tumor necrosis factor-

（TNF-

）， interleukin-6 （IL-6）， and interleukin-1

（IL-1

）. Tartrate-resistant acid phosphatase staining was used to detect the number of osteoclasts in the ankle joint. The expression of receptor activator of nuclear factor-

B ligand（RANKL） and osteoprotegerin（OPG） in ankle tissues was detected by immunohistochemistry（IHC）， real-time polymerase chain reaction （Real-time PCR）， and Western blot.

Result

Compared with the normal group， the model group showed decreased body weight （

0.01）， increased toe thickness and AI score （

0.01）， up-regulated serum levels of TNF-

，IL-6， and IL-1

（

0.01）， elevated number of osteoclasts （

0.01）， increased mRNA and protein expression of RANKL （

0.01）， reduced mRNA and protein expression of OPG （

0.05）， and increased RANKL/OPG ratio （

0.01）. Compared with the model group，the high-dose combination group showed increased body weight，decreased toe thickness and AI score， reduced mRNA and protein expression of RANKL， up-regulated mRNA and protein expression of OPG， and declining RANKL/OPG ratio （

0.05，

0.01）. The therapeutic effect was optimal in the high-dose combination group， followed by the low-dose combination group and the MTX group.

Conclusion

Zhiwang decoction combined with MTX can improve bone destruction induced by rheumatoid arthritis， which may be achieved by inhibiting the RANKL/OPG pathway. This study provides an experimental basis for the treatment of bone destruction induced by rheumatoid arthritis with integrated traditional Chinese and western medicine.

关键词

Keywords

references

中华医学会风湿病学分会 . 类风湿关节炎诊断及治疗指南［J］. 中华风湿病学杂志， 2010 ， 14 （ 4 ）： 265 - 270 .

曹芝艳，于泓 . 类风湿关节炎治疗研究进展［J］. 山东医药， 2019 ， 59 （ 21 ）： 108 - 111 .

SMOLEN J S ， ALETAHA D ， MCINNES I B . Rheumatoid arthritis ［J］. Lancet ， 2016 ， 388 （ 10055 ）： 2023 - 2038 .

SPARKS J A . Rheumatoid arthritis ［J］. Ann Intern Med ， 2019 ， 170 （ 1 ）： ITC1 - ITC16 .

马茹，陈冬雪，鄢泽然，等 . 补肾祛寒治尪汤联合甲氨蝶呤对大鼠胶原诱导关节炎模型的抗炎作用研究［J］. 上海中医药杂志， 2021 ， 55 （ 4 ）： 85 - 91 .

徐愿，阎小萍，肖诚，等 . 补肾祛寒治尪汤对胶原诱导关节炎大鼠抗炎与抗骨破坏的作用及对Wnt/ β -catenin通路的影响［J］. 北京中医药大学学报， 2020 ， 43 （ 4 ）： 289 - 295 .

何春晓，陈光耀，徐愿，等 . 补肾祛寒治尪汤联合甲氨蝶呤治疗类风湿关节炎系统综述［J］. 世界中西医结合杂志， 2018 ， 13 （ 12 ）： 1643 - 1647 .

TANAKA S . RANKL is a therapeutic target of bone destruction in rheumatoid arthritis ［J］. F1000Res ， 2019 ， doi： 10.12688/f1000research.17296.1 http://dx.doi.org/10.12688/f1000research.17296.1 .

TANAKA S ， TANAKA Y . RANKL as a therapeutic target of rheumatoid arthritis ［J］. J Bone Miner Metab ， 2021 ， 39 （ 1 ）： 106 - 112 .

YAN M ， SU J ， LI Y . Rheumatoid arthritis-associated bone erosions：evolving insights and promising therapeutic strategies ［J］. Biosci Trends ， 2020 ， 14 （ 5 ）： 342 - 348 .

ZHAO H ， XU H ， QIAO S ， et al . Boldine isolated from Litsea cubeba inhibits bone resorption by suppressing the osteoclast differentiation in collagen-induced arthritis ［J］. Int Immunopharmacol ， 2017 ， 51 ： 114 - 123 .

ZHOU Y ， WANG X ， AN Y ， et al . Disability and health-related quality of life in Chinese patients with rheumatoid arthritis：a cross-sectional study ［J］. Int J Rheum Dis ， 2018 ， 21 （ 9 ）： 1709 - 1715 .

WANG T ， HE C . TNF-α and IL-6：the link between immune and bone system ［J］. Curr Drug Targets ， 2020 ， 21 （ 3 ）： 213 - 227 .

KIKUTA J ， ISHII M . Bone imaging：osteoclast and osteoblast dynamics ［J］. Methods Mol Biol ， 2018 ， 1763 ： 1 - 9 .

TU Y ， WANG K ， TAN L ， et al . Dolichosin A，a coumestan isolated from Glycine tabacina ，inhibits IL-1 β -induced inflammation in SW982 human synovial cells and suppresses RANKL-induced osteoclastogenesis：from network pharmacology to experimental pharmacology ［J］. J Ethnopharmacol ， 2020 ， 258 ： 112855 .

YANG H ， LIU W ， ZHOU X ， et al . The association between RANK，RANKL and OPG gene polymorphisms and the risk of rheumatoid arthritis：a case-controlled study and Meta-analysis ［J］. Biosci Rep ， 2019 ， doi： 10.1042/BSR20182356 http://dx.doi.org/10.1042/BSR20182356 .

徐愿，陶庆文，王艳，等 . 基于伏邪学说辨治类风湿关节炎［J］. 北京中医药大学学报， 2018 ， 41 （ 3 ）： 181 - 185 .

王建明，张艳珍，马骁，等 . 补肾清化治尪汤联合甲氨喋呤治疗类风湿关节炎湿热伤肾证的疗效［J］. 中日友好医院学报， 2018 ， 32 （ 2 ）： 67 - 70 .

Views

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Pathogenesis and Traditional Chinese Medicine Treatment of Rheumatoid Arthritis Based on Theory of "Harmful Hyperactivity and Responding Inhibition"

Shire Biqing Pill in Treatment of Rheumatoid Arthritis with Damp-heat Obstruction Syndrome and Its Effect on OPG/RANKL/RANK Signaling Pathway and TNF-α

Effect of Aqueous Extract of Sophorae Tonkinensis Radix et Rhizoma on Bone Destruction and PI3K/Akt Signaling Pathway in Rheumatoid Arthritis

Research Progress of Role of Autophagy in Rheumatoid Arthritis and Traditional Chinese Medicine Intervention

Related Author

SONG Changyue

SU Xiaohui

WANG Jingbo

LI Siyu

CUI Huantian

KONG Xiangying

ZHOU Zhifeng

GUO Huiqing

Related Institution

Institute of Chinese Materia Media，China Academy of Chinese Medical Science

Yunnan University of Chinese Medicine

Zhengzhou Traditional Chinese Hospital of Orthopaedics

Henan Province Hospital of Traditional Chinese Medicine

Institute of Chinese Materia Medica，China Academy of Chinese Medical Sciences

AI问答

Postal code：100700
Tel：010-84076882 Email：syfjx_2010@188.com
Technical support is provided by Beijing Founder electronics co., LTD 京ICP备11006657号-10 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰