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]	Xue LUAN,Guanghai YAN,Haibo LI,Bo ZHANG,Wei ZHANG,Yuanyuan HUANG. Effect of salidroside on airway inflammation in mice with asthma and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2021, 47(3): 537-544.
[2]	Xiulan YE,Ming LI,Zhaojian WANG,Haiyan ZHAO,Jianning MA,Jing LIU,Yi ZHENG,Shunqing LI,Yu ZHANG. Effects of Cornus officinalis polysaccharide on behavior and multidrug resistance gene 1b and major vault protein expressions in hippocampus tissue of refractory epilepsy young rats [J]. Journal of Jilin University(Medicine Edition), 2021, 47(3): 644-651.
[3]	Shurong ZHANG,Qiying ZHANG. Regulatory effect of luteolin on Aspergillus fumigatus keratitis through TLR4/MyD88 signaling pathway in rats [J]. Journal of Jilin University(Medicine Edition), 2021, 47(2): 315-322.
[4]	Zhenjie MA,Lan MA,Zhen JIA. Effect of celiac plexus block on stress response and immune inflammation of rats after partial hepatectomy and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2021, 47(2): 407-413.
[5]	Jie WU,Xuehua YANG,Ling MA,Wenqiang FAN,Dongdong FU,Xiao GAO,Shufei ZUO,Shu LIANG,Yilu QIN,Peishan WANG,Jinyan GUO. Effect of miR-26a-5p on apoptosis of human rheumatoid arthritis fibroblast-like synovial cells through JAK2/STAT3 signaling pathway [J]. Journal of Jilin University(Medicine Edition), 2021, 47(2): 460-468.
[6]	QIAO Liangwei, QU Qingshan, LI Ming. Effect of LncRNA-ATB on acute rejection of kidney transplanted rats by regulating miR-200c expression [J]. Journal of Jilin University(Medicine Edition), 2020, 46(05): 955-962.
[7]	ZHANG Yiqun, JIANG Weibo, YUAN Sheng, YU Wei. Analysis on follow-up results of postoperative function and recurrence of 23 patients with tenosynovial giant cell tumor in hand [J]. Journal of Jilin University(Medicine Edition), 2020, 46(05): 1061-1064.
[8]	ZHAO Liping, HUANG Shubing, ZHANG Boping, ZHOU Zhilan, JIA Xuebing, SUN Mengfei, QIAO Chenmeng, CHEN Xue, SHEN Yanqin, CUI Chun. Inhibitory effect of Lactobacillus rhamnosus on intestinal inflammation after spinal cord injury in zebrafishes and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2020, 46(04): 680-686.
[9]	WANG Yingying, SUN Lijuan, FAN Ziwei, GU Hong, YOU Xianmei, GUAN Tianhao, ZHANG Chengyi, CHEN Xi. Inhibitory effect of baicalin on autophagy of synovial RSC-364 cells of rats induced by lipopolysaccharide [J]. Journal of Jilin University(Medicine Edition), 2020, 46(03): 498-503.
[10]	LIU Yiping, SUN Xiaolin, NI Shilei, ZHAO Cong, GE Yue, ZHOU Yanmin, LI Minghe. Synovial chondromatosis of temporomandibular joint with disc perforation: A case report and literature review [J]. Journal of Jilin University(Medicine Edition), 2020, 46(03): 639-642.
[11]	WANG Yazhou, HE Peng, WANG Danhong. Effects of montelukast on proliferation and apoptosis of airway smooth muscle cells in asthmatic rats by inhibiting TLR4/NF-κB signaling pathway [J]. Journal of Jilin University(Medicine Edition), 2020, 46(02): 274-279.
[12]	WAN Qi, YU Baogang. Effects of miR-125b on proliferation and migration of cardiac fibroblasts by TLR4/NF-κB signaling pathway [J]. Journal of Jilin University(Medicine Edition), 2020, 46(02): 286-291.
[13]	ZHAO Miao, WANG Yi, ZHANG Ying, FENG Yumei, CAO Yawen, JIANG Haisen, LI Wei. Improvement effect of curcumin on cognitive function in mice with sleep deprivation and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2019, 45(06): 1373-1378.
[14]	WANG Tianyue, ZHOU Qianlan, SHANG Yunxiao. Effect of epigallocatechin-3-gallate on airway inflammation and Treg/Th17 immune balance of mice with obese asthma [J]. Journal of Jilin University(Medicine Edition), 2019, 45(03): 491-497.
[15]	CHEN Xi, BAI Lin, WANG Yingying, CAO Shuang, MU Hongping, WANG Yilin, SHI Hao, ZHANG Qian, GAO Xin, ZHANG Chengyi, ZHANG Ruowen. Inhibitory effect of tanshinone ⅡA on proliferation and migration of human liver cancer HepG2 cells and its apoptosis-promoting effect [J]. Journal of Jilin University(Medicine Edition), 2019, 45(03): 531-538.