白芍总苷介导TLR4/NF-κB信号通路对类风湿关节炎模型大鼠的治疗作用及其机制

doi:10.13481/j.1671-587X.20210219

Abstract

Abstract: Objective

To investigate the effects of total glucosides of paeony （TGP） on joint condition， inflammatory factors and apoptosis factors in the rheumatoid arthritis （RA） rats by regulating the Toll-like receptor 4（TLR4） /nuclear factor-κB（NF-κB） signaling pathway， and to clarify its mechanism.

Methods

The collagen-induced arthritis （CIA） rat models were established with bovine type Ⅱ collagen and complete Freund’s adjuvant， and the CIA model rats were randomly divided into CIA group， low dose （25 mg·kg^－1）of TGP（TGP-LD） group， middle dose （50 mg·kg^－1） of TGP（TGP-MD） group， and high dose （100 mg·kg^－1） of TGP（TGP-HD）， with 10 rats in each group.Another 10 healthy SD rats were selected as control group . The body weights， arthritis indexes and joint swelling degrees of the rats in each group were measured， and the levels of tumor necrosis factor-α （TNF-α），interleukin-6 （IL-6） and interleukin-1β （IL-1β） in serum of the rats in each group were detected by ELISA method. The mRNA and protein expression levels of TLR4， NF-κB， Cleaved caspase-3， B cell lymphoma -2 （Bcl-2） and Bcl-2 associated X protein （Bax） in joint synovium tissue of the rats in each group were detected by Real-time fluorescence quantitative PCR（RT-qPCR） and Western blotting methods.

Results

Compared with control group， the body weight and the expression levels of Cleaved caspase-3 and Bax mRNA and proteins in joint synovium tissue of the rats in CIA group were significantly decreased （P<0.05）， while the arthritis index， the joint swelling degree， the expression levels of Bcl-2， TLR4，and NF-κB p65 mRNA and proteins in joint synovium tissue， and the TNF-α， IL-6， and IL-1β levels in serum of the rats in CIA group were significantly increased （P<0.05）. Compared with CIA group，the body weights and the expression levels of Cleaved caspase-3 and Bax mRNA and protein in joint synovium tissue of the rats in different doses of TGP groups were significantly increased（P<0.05）， while the arthritis indexes，the joint swelling degrees， the expression levels of Bcl-2， TLR4，and NF-κB p65 mRNA and proteins in joint synovium tissue， and the TNF-α， IL-6， IL-1β levels in serum of the rats were significantly decreased（P<0.05）.Compared with TGP-LD group，the body weights and the expression levels of Cleaved caspase-3 and Bax mRNA and proteins in joint synovium tissue of the rats in TGP-MD group and TGP-HD group were significantly increased（P<0.05），while the arthritis indexes， the joint swelling degrees， the expression levels of TLR4，NF-κB，p65 and Bcl-2 in joint synovium tissue， and the TNF-α， IL-6， and IL-1β levels in serum of the rats were significantly decreased（P<0.05）.

Conclusion

TGP can improve the symptoms of CIA， reduce the inflammatory reaction and promote the apoptosis of synovial tissue by inhibiting TLR4/NF- κB signal pathway.

Key words: total glucoside of paeony, Toll-like receptor 4, nuclear factor-κB, rheumatoid arthritis, inflammation, synovial

CLC Number:

R684.3

Lijun YAN,Shengquan TONG,Jing LIU,Dongmei GAO,Nanfang CHEN,Jie HU. Therapeutic effect of total glucosides of paeony in model rats with rheumatoid arthritis by mediating TLR4/NF-κB signaling pathway and its mechanisim[J].Journal of Jilin University(Medicine Edition), 2021, 47(2): 390-396.

Figures/Tables 8

Tab. 1

Fig. 1

Tab. 1

Tab. 2

Tab. 3

Fig. 2

Tab. 4

Fig.3

References 22

1	KLOOSTER P M，OUDE VOSHAAR M A H，FAKHOURI W， et al. Long-term clinical， functional， and cost outcomes for early rheumatoid arthritis patients who did or did not achieve early remission in a real-world treat-to-target strategy［J］.Clin Rheumatol，2019，38（10）：2727-2736.
2	GOU K J， ZENG R， REN X D， et al. Anti-rheumatoid arthritis effects in adjuvant-induced arthritis in rats and molecular docking studies of Polygonum orientale L. extracts［J］. Immunol Lett， 2018， 201： 59-69.
3	TENTEN-DIEPENMAAT M， DEKKER J， HEYMANS M W， et al. Systematic review on the comparative effectiveness of foot orthoses in patients with rheumatoid arthritis［J］. J Foot Ankle Res， 2019， 12： 32.
4	XU N， WANG Y， ZHAO S， et al. Naltrexone （NTX） relieves inflammation in the collagen-induced- arthritis （CIA） rat models through regulating TLR4/NFκB signaling pathway［J］. Int Immunopharmacol， 2020， 79： 106056.
5	YAN S， WANG P， WANG J H， et al. Long non-coding RNA HIX003209 promotes inflammation by sponging miR-6089 via TLR4/NF-κB signaling pathway in rheumatoid arthritis［J］. Front Immunol， 2019， 10： 2218.
6	LIU W， WU Y H， ZHANG L， et al. MicroRNA-146a suppresses rheumatoid arthritis fibroblast-like synoviocytes proliferation and inflammatory responses by inhibiting the TLR4/NF-kB signaling［J］. Oncotarget， 2018， 9（35）： 23944-23959.
7	JIA X Y， CHANG Y， SUN X J， et al. Total glucosides of paeony inhibit the proliferation of fibroblast-like synoviocytes through the regulation of G proteins in rats with collagen-induced arthritis［J］.Int Immunopharmacol， 2014， 18（1）： 1-6.
8	LI H， CAO X Y， DANG W Z， et al. Total Glucosides of Paeony protects against collagen-induced mouse arthritis via inhibiting follicular helper T cell differentiation［J］. Phytomedicine， 2019， 65： 153091.
9	LEI Z H， OUYANG L， GONG Y， et al. Effect of eriodictyol on collagen-induced arthritis in rats by Akt/HIF-1α pathway［J］. Drug Des Devel Ther， 2020， 14： 1633-1639.
10	HUGHES C， WOLOS J A， GIANNINI E H， et al. Induction of T helper cell hyporesponsiveness in an experimental model of autoimmunity by using nonmitogenic anti-CD3 monoclonal antibody［J］.J Immunol， 1994，153（7）：3319-3325.
11	马潇苒，李无阴.中药治疗类风湿关节炎的研究进展［J］.风湿病与关节炎， 2018， 7（10）： 66-70.
12	李丽萍，张杜娟，冯坤苗.白芍总苷治疗类风湿关节炎药理作用及机制研究进展［J］.山东医药，2019，59（26）：111-113.
13	李宗祥，肖凯. 基于TLR4/NF-κB信号通路探讨通痹胶囊对胶原诱导型关节炎治疗机制的研究［J］. 中国免疫学杂志， 2019， 35（12）：1453-1457.
14	赵莉，苏双全，夏林，等. 白芍总苷对糖尿病大鼠肾组织TLR2、TLR4/NF-κB信号通路调节的影响［J］. 安徽医科大学学报， 2012， 47（5）：34-38.
15	POLLARD K M， ESCALANTE G M， HUANG H， et al. Induction of systemic autoimmunity by a xenobiotic requires endosomal TLR trafficking and signaling from the late endosome and endolysosome but not type Ⅰ IFN［J］. J Immunol， 2017， 199（11）： 3739-3747.
16	DIAKOWSKA D， NIENARTOWICZ M， GRABOWSKI K， et al. Toll-like receptors TLR-2， TLR-4， TLR-7， and TLR-9 in tumor tissue and serum of the patients with esophageal squamous cell carcinoma and gastro-esophageal junction cancer［J］. Adv Clin Exp Med， 2019， 28（4）： 515-522.
17	XU D， YAN S， WANG H， et al. IL-29 Enhances LPS/TLR4 mediated inflammation in rheumatoid arthritis［J］. Cell Physiol Biochem， 2015， 37（1）：27-34.
18	蔡辉，张群燕，郭郡浩，等. 类风湿关节炎患者外周血血浆中TNF-α、IL-1β及IL-10的表达及临床意义［J］.中华中医药学刊，2017，35（3）：519-521.
19	徐晓玲，梁瑞霞，黄君健，等. 细胞因子在类风湿关节炎发病机制中的作用［J］. 生物技术通讯，2008，19（1）：111-114.
20	王霖，王文杰. 破骨细胞在类风湿性关节炎骨破坏中的作用及其调控机制［J］. 生理科学进展， 2004， 35（3）：269-272.
21	QUE W， LIU H， YANG Q， et al. NK4 inhibits the proliferation and induces apoptosis of human rheumatoid arthritis synovial cells［J］. Cell Biochem Funct， 2018， 36（5）： 273-279.
22	范文强,马玲,吴洁,等.miR-708-5p对类风湿关节炎滑膜成纤维细胞凋亡、炎症因子分泌和TLR4/NF-κB信号通路的影响[J].郑州大学学报(医学版),2020,55(5):705-710.

Primer	Sequence (5'－3')
NF-κB p65	Forward:ACGATCTGTTTCCCCTCATC
NF-κB p65	Reserse: TGCTTCTCTCCCCAGGAATA
TLR4	Forward:TTGCCTTCATTACAGGGACTT
TLR4	Reserse: AGATACCGGTGGAGGCTGACT
Cleaved caspase-3	Forward: AGATGGTTTGAGCCTGAGCA
Cleaved caspase-3	Reserse: CAGTGCGTATGGAGAAATGG
Bax	Forward: GAGACACCTGAGCTGACCTTG
Bax	Reserse: GCTCCATGTTGTTGTCCAGTT
Bcl-2	Forward: GGTGAACTGGGGGAGGATTG
Bcl-2	Reserse: AGAGCGATGTTGTCCACCAG
β-actin	Forward:GCCATGTACGTAGCCATCCA
	Reserse: GAACCGCTCATTGCCGATAG

Group	TNF-α	IL-6	IL-1β
Control	49.31±4.82	28.64±3.06	21.17±1.29
CIA	107.63±16.29^*	83.43±10.02^*	68.35±9.63^*
TGP-LD	79.26±8.20^△	64.57±7.79^△	57.68±6.26^△
TGP-MD	64.11±7.13^△#	60.32±7.25^△#	42.44±5.03^△#
TGP-HD	58.20±5.61^△#	42.64±5.87^△#	37.29±2.51^△#

Group	Cleaved caspase-3	Bax	Bcl-2
Control	1.00±0.02	0.99±0.04	1.01±0.03
CIA	0.41±0.05^*	0.35±0.05^*	2.95±0.24^*
TGP-LD	0.56±0.07^△	0.54±0.07^△	2.46±0.17^△
TGP-MD	0.75±0.04^△#	0.63±0.05^△#	2.07±0.15^△#
TGP-HD	0.86±0.06^△#	0.82±0.09^△#	1.69±0.18^△#

Group	Cleaved caspase-3	Bax	Bcl-2
Control	0.63±0.05	0.66±0.08	0.82±0.07
CIA	0.38±0.04^*	0.26±0.04^*	1.98±0.32^*
TGP-LD	0.47±0.06^△	0.41±0.02^△	1.67±0.21^△
TGP-MD	0.60±0.09^△#	0.47±0.04^△#	1.34±0.16^△#
TGP-HD	0.69±0.12^△#	0.56±0.07^△#	0.97±0.12^△#

Group	TLR4 mRNA	NF-κB p65 mRNA	TLR4 protein	NF-κB p65 protein
Control	1.01±0.03	1.00±0.05	0.52±0.08	0.31±0.04
CIA	3.65±0.38^*	2.78±0.21^*	1.02±0.11^*	0.78±0.06^*
TGP-LD	3.18±0.29^△	2.25±0.23^△	0.89±0.08^△	0.63±0.05^△
TGP-MD	2.74±0.20^△#	1.98±0.17^△#	0.77±0.05^△#	0.55±0.05^△#
TGP-HD	1.73±0.15^△#	1.36±0.11^△#	0.48±0.04^△#	0.35±0.04^△#

Therapeutic effect of total glucosides of paeony in model rats with rheumatoid arthritis by mediating TLR4/NF-κB signaling pathway and its mechanisim

RichHTML

PDF (PC)

Like

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 8

References 22

Related Articles 15

Metrics

Comments

Recommended 10

[1]	Lina XU,Xiaoshuang HE,Wenyan XIN,Chao WU. Improvement effect of bone marrow mesenchymal stem cell-derived exosomes on airway resistance and lung tissue lesion in mice of asthmatic models and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2026, 52(3): 602-611.
[2]	Lingyu WANG,Ruili LI,Yu ZHANG,Meitong GUO,Yalong LIANG,Xu HAN,Xiaowei HUANG,Yang ZHANG,Xiuhua YU. Improvement effect of Renshen Jianzhi Tablets on learning and memory ability in cognitive impairment model rats and its protective effect on hippocampal neurons and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2026, 52(3): 719-729.
[3]	Jingyuan WANG,Fang CHEN,Yancun LIU,Shixin LI,Songtao SHOU. Protective effect of silencing coagulation factor V gene on septic rats by inhibiting JNK1/2 and p38 MAPK signaling pathways [J]. Journal of Jilin University(Medicine Edition), 2026, 52(2): 418-428.
[4]	Aimei LI,Jingfei HAN,Li DENG,Siyu CHEN. Protective effect of dexmedetomidine on myocardial injury in myocardial ischemia/reperfusion injury model rats [J]. Journal of Jilin University(Medicine Edition), 2026, 52(2): 451-459.
[5]	Liyang BAI,Di MA,Shuo HUANG,Chuan WANG,Jiaxin LIANG,Tian WANG,Liuping CUI,Lingmin ZHAO,Meizhen XIE,Mengyue YAO,Ying CHEN,Lijuan WANG. Effect of carotid plaque composition and HDL-C levels on symptomatic carotid artery stenosis [J]. Journal of Jilin University(Medicine Edition), 2026, 52(2): 491-498.
[6]	Huiyan ZHU,Min CHEN,Jinxian LI,Chunli LI. Effect of enriched environment on neurofunctional damage in rats with ischemic stroke via transcription factor EB-mediated autophagy [J]. Journal of Jilin University(Medicine Edition), 2026, 52(1): 116-124.
[7]	Sitong CHEN,Dan YANG,Qingjie LI,Xiaolei TANG,Han WANG,Yangyang LIU,Tiejun LIU. Ameliorative effect of puerarin on non-alcoholic fatty liver disease induced by high-fat diet in mice and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2026, 52(1): 44-55.
[8]	Sifan FENG,Yunfeng LI,Jiaying WANG,Fubin MA,Yan WANG. Effects of heme-binding protein 1 gene knockdown on proliferation, migration, and inflammatory response of microglia BV2 and their mechanisms [J]. Journal of Jilin University(Medicine Edition), 2025, 51(6): 1532-1541.
[9]	Tao SUN,Zhiyin DAI,Xuan LI,Chaopu ZHANG,Shu DING,Jianwei ZHAO. Predictive value of pan-immune-inflammation index for major adverse cardiovascular events within 1 year after PCI in elderly patients with coronary heart disease [J]. Journal of Jilin University(Medicine Edition), 2025, 51(6): 1655-1660.
[10]	Ying HU,Yong HUANG. Improvement effect of short-chain fatty acids on inflammation-induced autophagic damage in ovarian granulosa cells in polycystic ovary syndrome and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2025, 51(5): 1340-1348.
[11]	Guang YANG,Zhifang ZHENG,Xinhua ZHANG. Research progress in protective effect of miRNA on neonatal hypoxic-ischemic brain injury [J]. Journal of Jilin University(Medicine Edition), 2025, 51(5): 1423-1428.
[12]	Cheng CHEN,Jingyao LI,Wanxiang HU,Donghui LIU,Zhihong CHEN. Protective effect of sericin on streptozotocin-induced INS-1 cell damage by regulating PI3K/Akt/NF-κB signaling pathway through Akt1 and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2025, 51(3): 590-598.
[13]	Xingxiang WANG,Ying ZHAO,Qiaotong REN,Hefei WANG,Gang PU,Chun LI. Promotive effect of M2 macrophages on epithelial-mesenchymal transition and cisplatin resistance in non-small cell lung cancer A549 cells by regulating NF-κB signaling pathway [J]. Journal of Jilin University(Medicine Edition), 2025, 51(3): 642-652.
[14]	Junjie JIANG,Hao WU,Kang HE,Zhiqiang SAN,Qing YANG,Hui LI,Na LI. Repair effect of ginseng polypeptide thermosensitive hydrogel on heat-induced skin injury in rats and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2025, 51(2): 360-369.
[15]	Xiaoxia HU,Yalong LI,Dongliang YANG,Bazeren LA,Xinyue LIU. Effect of high glucose on polarization of Raw264.7 macrophages in vitro [J]. Journal of Jilin University(Medicine Edition), 2025, 51(2): 403-411.