Effect of Shikonin on Synovitis in Mice with Collagen-induced Arthritis Through MAPK1/MAPK Pathway

TONG Wenting; HE Lianhua

doi:10.13422/j.cnki.syfjx.20231138

您当前的位置：

首页 >

文章列表页 >

Effect of Shikonin on Synovitis in Mice with Collagen-induced Arthritis Through MAPK1/MAPK Pathway

更新时间：2023-10-20

- Effect of Shikonin on Synovitis in Mice with Collagen-induced Arthritis Through MAPK1/MAPK Pathway
  增强出版
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 29, Issue 22, Pages: 56-63(2023)
- 作者机构：
  
  1.赣南卫生健康职业学院，江西赣州 341000
  2.深圳市中医院/广州中医药大学第四临床医学院，广东深圳 518033
  3.深圳华润三九医药贸易有限公司，广东深圳 518109
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20231138
  CLC： R2-0;R33;R593.22;R289
- Received：30 June 2023，
  
  Online First：30 August 2023，
  
  Published：20 November 2023
- 稿件说明：
移动端阅览
童文婷,何莲花.紫草素通过MAPK1/MAPK通路对小鼠胶原诱导型关节炎中滑膜炎症的影响[J].中国实验方剂学杂志,2023,29(22):56-63.

TONG Wenting,HE Lianhua.Effect of Shikonin on Synovitis in Mice with Collagen-induced Arthritis Through MAPK1/MAPK Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(22):56-63.
童文婷,何莲花.紫草素通过MAPK1/MAPK通路对小鼠胶原诱导型关节炎中滑膜炎症的影响[J].中国实验方剂学杂志,2023,29(22):56-63. DOI： 10.13422/j.cnki.syfjx.20231138.

TONG Wenting,HE Lianhua.Effect of Shikonin on Synovitis in Mice with Collagen-induced Arthritis Through MAPK1/MAPK Pathway[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(22):56-63. DOI： 10.13422/j.cnki.syfjx.20231138.

摘要

目的

探究紫草素对DBA/1小鼠胶原诱导型关节炎（CIA）中滑膜炎的影响。

方法

将36只DBA/1小鼠按体质量随机分为6组，每组6只。6组分别为正常组、CIA组，紫草素1、2、4 mg·kg

-1

组及甲氨蝶呤（MTX）0.5 mg·kg

-1

组。CIA组、紫草素低、中、高剂量组及MTX组在第1天以等体积牛Ⅱ型胶原和完全弗氏佐剂免疫DBA/1小鼠，于第21天以等体积牛Ⅱ型胶原和不完全弗氏佐剂二次免疫DBA/1小鼠建立CIA模型，并于二次免疫当天开始灌胃给药，除MTX为每周3次外，其余每天1次，共给药28 d。第22天开始观察CIA小鼠的关节红肿等症状并进行关节炎评分，第49天取材后，采用苏木素-伊红（HE）染色进行组织病理学检查观察CIA小鼠关节滑膜炎症的情况；免疫荧光（IF）双标法检测CIA小鼠关节滑膜中波形蛋白（Vimentin）和丝裂原活化蛋白激酶1（MAPK1）的表达；网络药理学预测出紫草素作用于RA的靶标是MAPK1，采用分子对接方式进行验证；蛋白免疫印迹法（Western blot）检测小鼠关节滑膜中细胞外调节蛋白激酶（ERK）、c-Jun氨基末端激酶（JNK）、p38、磷酸化（p）-ERK、p-JNK、p-p38蛋白表达。

结果

与正常组比较，CIA组小鼠关节炎评分、发病率及滑膜炎症情况显著升高，与正常组比较，CIA组小鼠关节结构严重紊乱，关节软骨及骨破坏明显，骨侵蚀严重（

0.01）；小鼠关节滑膜中Vimentin及MAPK1的表达明显增多；小鼠关节滑膜中p-ERK/ERK，p-JNK/JNK及p-p38/p38的蛋白表达显著增高（

0.01）；与CIA组比较，紫草素各剂量组和MTX组小鼠关节结构均相对正常，骨侵蚀、骨破坏较轻，关节面相对完整光滑；分子对接结果证明紫草素的作用靶点为MAPK1；紫草素各剂量组和MTX组关节滑膜中Vimentin及MAPK1表达显著降低（

0.01），小鼠关节滑膜中p-ERK/ERK，p-JNK/JNK及p-p38/p38的蛋白表达显著降低（

0.01）。

结论

紫草素可靶向MAPK1抑制CIA小鼠中p-ERK，p-JNK及p-p38的蛋白表达，进而治疗RA。

Abstract

Objective

To observe the effect of shikonin （SKN） on synovitis in DBA/1 mice with collagen-induced arthritis （CIA）.

Method

Thirty-six DBA/1 mice were randomly divided into a normal group， a CIA group， low-， medium-， and high-dose SKN groups （1， 2， and 4 mg·kg

-1

）， and a methotrexate （MTX， 0.5 mg·kg

-1

） group， with 6 mice in each group. Mice in the CIA group， the SKN groups， and the MTX group were immunized with an equal volume of bovine type Ⅱ collagen and complete Freund's adjuvant on day 1. On day 21， those mice received a second immunization with an equal volume of bovine type Ⅱ collagen and incomplete Freund's adjuvant to establish the CIA model. On the day of the second immunization， mice were treated with drugs by gavage. Mice in the MTX group received oral administration three times a week， while others received once per day， for 28 days. On day 22， the symptoms of arthritis， such as redness and swelling of joints， in CIA mice were observed， and arthritis scores were recorded. On day 49 after sample collation， histopathological examination of synovial inflammation in CIA mice was performed using hematoxylin-eosin （HE） staining. Immunofluorescence （IF） double labeling was used to detect the expression of vimentin and mitogen-activated protein kinase 1 （MAPK1） in the synovium of CIA mice. Network pharmacology predicted that the target of SKN in rheumatoid arthritis （RA） was MAPK1， which was verified by molecular docking. Western blot was used to detect the expression of extracellular signal-regulated kinase （ERK）， c-Jun N-terminal kinase （JNK）， p38， phosphorylated （p）-ERK， p-JNK， and p-p38 proteins in the synovial membrane of mice.

Result

Compared with the normal group， the CIA group showed significantly higher arthritis scores， morbidity， and synovial inflammation， severely disrupted joint structure， evident articular cartilage and bone destruction， severe bone erosion （

0.01）， increased expression of vimentin and MAPK1 in the synovium of mice， and increased protein expression of p-ERK/ERK， p-JNK/JNK， and p-p38/p38 in the synovium of mice （

0.01）. Compared with the CIA group， the SKN groups and the MTX group showed relatively normal joint structure， with milder bone erosion and bone destruction， and smoother articular surfaces. Molecular docking results confirmed that the target of SKN was MAPK1. In the SKN groups and the MTX group， the expression of vimentin and MAPK1 in the synovial membrane was significantly reduced （

0.01）， and the protein expression of p-ERK/ERK， p-JNK/JNK， and p-p38/p38 in the synovium of mice was significantly reduced （

0.01）.

Conclusion

SKN can target MAPK1 to inhibit the protein expression of p-ERK， p-JNK， and p-p38 in CIA mice， thereby treating RA.

关键词

Keywords

references

DING Q ， HU W ， WANG R ， et al . Signaling pathways in rheumatoid arthritis：Implications for targeted therapy ［J］. Signal Transduct Target Ther ， 2023 ， 8 （ 1 ）： 68 .

YAP H Y ， TEE S Z ， WONG M M ， et al . Pathogenic role of immune cells in rheumatoid arthritis： Implications in clinical treatment and biomarker development ［J］. Cells ， 2018 ， doi： 10.3390/cells7100161 http://dx.doi.org/10.3390/cells7100161 .

HU H ， LUAN L ， YANG K ， et al . Burden of rheumatoid arthritis from a societal perspective：A prevalence-based study on cost of this illness for patients in China ［J］. Int J Rheum Dis ， 2018 ， 21 （ 8 ）： 1572 - 1580 .

马生军，耿阳，马露，等 . 药用紫草研究进展［J］. 中国现代中药， 2021 ， 23 （ 1 ）： 177 - 184 .

何莲花，汪倩倩，孙丛丛，等 . 紫草素对类风湿关节炎成纤维样滑膜细胞功能的影响［J］. 中国中药杂志， 2020 ， 45 （ 19 ）： 4712 - 4718 .

HE L ， LUAN H ， HE J ， et al . Shikonin attenuates rheumatoid arthritis by targeting SOCS1/JAK/STAT signaling pathway of fibroblast like synoviocytes ［J］. Chin Med ， 2021 ， 16 （ 1 ）： 96 .

RALPH J A ， MORAND E F . MAPK phosphatases as novel targets for rheumatoid arthritis ［J］. Expert Opin Ther Targets ， 2008 ， 12 （ 7 ）： 795 - 808 .

何莲花，栾慧杰，何娟，等 . 二妙散对DBA/1小鼠胶原诱导型关节炎中Th17/Treg细胞分化的影响［J］. 中国实验方剂学杂志， 2023 ， 29 （ 2 ）： 66 - 72 .

何莲花，栾慧杰，单宏颖，等 . 防己黄芪汤对DBA/1小鼠胶原诱导型关节炎及血管新生的影响［J］. 中国实验方剂学杂志， 2021 ， 27 （ 17 ）： 16 - 23 .

栾慧杰，何莲花，何娟，等 . 积雪草苷对DBA/1小鼠胶原诱导型关节炎中Th17/Treg细胞表达的影响［J］. 中国实验方剂学杂志， 2022 ， 28 （ 4 ）： 76 - 83 .

KIM Y O ， HONG S J ， YIM S V . The efficacy of shikonin on cartilage protection in a mouse model of rheumatoid arthritis ［J］. Korean J Physiol Pharmacol ， 2010 ， 14 （ 4 ）： 199 - 204 .

PRADO C ， UGALDE V ， GONZÁLEZ H ， et al . STAT3 activation in combination with NF- κ B inhibition induces tolerogenic dendritic cells with high therapeutic potential to attenuate collagen-induced arthritis ［J］. J Immunol Res ， 2019 ， 2019 ： 1982570 .

杨哲，李明强，纪超凡，等 . 滑膜细胞代谢在类风湿关节炎中作用的研究进展［J］. 中国临床医学， 2019 ， 26 （ 5 ）： 787 - 790 .

白永峰，曹乌吉斯古楞 . 滑膜成纤维细胞在类风湿性关节炎中的作用及中药干预研究进展［J］. 中国实验方剂学杂志， 2022 ， 28 （ 14 ）： 226 - 234 .

宋腾，周亚琪，周广玺，等 . 紫草素在免疫相关性疾病中作用的研究进展［J］. 中国医药， 2023 ， 18 （ 4 ）： 629 - 632 .

LIU C ， HE L ， WANG J ， et al . Anti-angiogenic effect of Shikonin in rheumatoid arthritis by downregulating PI3K/Akt and MAPKs signaling pathways ［J］. J Ethnopharmacol ， 2020 ， 260 ： 113039 .

LIANHUA H E ， HUIJIE L ， QINGXIA Q ， et al . Shikonin alleviates collagen-induced arthritis mice by inhibiting M1 macrophage polarization ［J］. J Tradit Chin Med ， 2022 ， 42 （ 6 ）： 932 - 939 .

LI J ， PANG J ， LIU Z ， et al . Shikonin induces programmed death of fibroblast synovial cells in rheumatoid arthritis by inhibiting energy pathways ［J］. Sci Rep ， 2021 ， 11 （ 1 ）： 18263 .

DAI Q ， FANG J ， ZHANG F S . Dual role of shikonin in early and late stages of collagen type Ⅱ arthritis ［J］. Mol Biol Rep ， 2009 ， 36 （ 6 ）： 1597 - 1604 .

YANG K Y ， CHEN D L . Shikonin inhibits inflammatory response in rheumatoid arthritis synovial fibroblasts via lncRNA-NR024118 ［J］. Evid Based Complement Alternat Med ， 2015 ， 2015 ： 631737 .

SUN W X ， LIU Y ， ZHOU W ， et al . Shikonin inhibits TNF- α production through suppressing PKC-NF- κ B-dependent decrease of IL-10 in rheumatoid arthritis-like cell model ［J］. J Nat Med ， 2017 ， 71 （ 2 ）： 349 - 356 .

LIU Z ， LU J ， FANG H ， et al . m6A Modification-mediated DUXAP8 regulation of malignant phenotype and chemotherapy resistance of hepatocellular carcinoma through miR-584-5p/MAPK1/ERK pathway axis ［J］. Front Cell Dev Biol ， 2021 ， 9 ： 783385 .

SI H ， ZHANG N ， SHI C ， et al . Tumor-suppressive miR-29c binds to MAPK1 inhibiting the ERK/MAPK pathway in pancreatic cancer ［J］. Clin Transl Oncol ， 2023 ， 25 （ 3 ）： 803 - 816 .

甘苡榕，桂雄斌，俞渊，等 . 大黄灵仙方调控TAK1与TRAF6相互作用及共定位对胆管细胞炎症反应的影响［J］. 中国实验方剂学杂志， 2022 ， 28 （ 6 ）： 92 - 99 .

KASHATUS J A ， NASCIMENTO A ， MYERS L J ， et al . ERK2 phosphorylation of DRP1 promotes mitochondrial fission and MAPK-driven tumor growth ［J］. Mol Cell ， 2015 ， 57 （ 3 ）： 537 - 551 .

Views

277

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

Modified Guishenwan Alleviates Inflammation in Rat Model of Polycystic Ovary Syndrome by Inhibiting MAPK/NF-κB Pathway

Shikonin Inhibits Inflammation of Psoriasis Cell Model by Regulating cGAS/STING Signaling Pathway

Traditional Chinese Medicine Treats Tumors via MAPK/NF-κB Signaling Pathway： A Review

Role of PPARγ in Ulcerative Colitis and Traditional Chinese Medicine Intervention： A Review

Clinical and Mechanistic Research Progress on Classical Traditional Chinese Medicine Formulas in Treatment of Chronic Heart Failure

Related Author

TIAN Jiayu

QIN Wenyi

YANG Juan

RONG Xiaofeng

LYU Chong

QIAO Xianhua

GAO Juanjuan

TIAN Fei

Related Institution

The First Affiliated Hospital of Chongqing Medical University

Hebei University of Chinese Medicine

Xingtai People's Hospital

Shandong University of Traditional Chinese Medicine（TCM）

Yantai Hospital of TCM

AI问答

Address：No. 16, Nanxiaojie, Dongzhimennei, Beijing 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