薯蓣皂苷对滑膜炎大鼠症状的改善作用和对TLR2-NF-κB信号通路的调节作用及其机制

doi:10.13481/j.1671-587X.20210417

Abstract

Abstract: Objective

To investigate the improvement effect of diosgenin on the symptoms of synovitis rats and the influence in Toll-like receptor 2 （TLR2） - nuclear factor kappaB（NF-κB） signaling pathway， and to elucidate the possible mechanism of diosgenin in the treatment of synovitis.

Methods

The synovitis models were established by intradermal injection of Freund’s complete adjuvant，and the model rats were randomly divided into model group（n=10），positive drug group （indomethacin 10 mg·kg^－1）（n=11）， low dose of diosgenin group （60 mg·kg^－1）（n=12）， and high dose of diosgenin group （120 mg·kg^－1）（n=12）； another 10 healthy rats were set as control group. Joint pain scoreing and water volume methods were used to evaluate the changes of joint pain and swelling on the 3rd， 7th， 14th and 21st days after intervention. The levels of tumor necrosis factor-α （TNF-α） and interleukin-1β （IL-1β） in synovial tissue of the rats in various groups were detected by enzyme-linked immunosorbent assay method. HE staining was used to observe the pathomorphology of synovium tissue of the rats in various groups. Western bloting method was used to detect the expression levels of TLR2， MyD88， NF-κB and p-NF-κB p65 proteins in synovium tissue of the rats in various groups.

Results

Compared with control group， the pain scores of the rats in model group， positive drug group， low dose of diosgenin group and high dose of diosgenin group at different time points were significantly increased P<0.05）， the degrees of toe swelling were significantly increased （P<0.05）， and the levels of TNF-α and IL-1β in synovial tissue were significantly increased （P<0.05）. Compared with model group， the pain scores of the rats in positive drug group， low dose of diosgenin group and high dose of diosgenin group at different time points were significantly decreased （P<0.05）， the degrees of toe swelling were significantly decreased （P<0.05）， and the levels of TNF-α and IL-1β in synovial tissue were significantly decreased （P<0.05）.Compared with positive drug group， the pain scores of the rats in low dose of diosgenin group and high dose of diosgenin group at different time points were significantly increased （P<0.05）， the degrees of toe swelling were significantly increased （P<0.05），and the levels of TNF-α and IL-1β in synovial tissue were significantly increased （P<0.05）. Compared with low dose of diosgenin group， the pain score of the rats in high dose of diosgenin group was significantly decreased（P<0.05）， the degree of toe swelling was significantly decreased （P<0.05）， and the levels of TNF-α and IL-1β in synovial tissue were significantly decreased （P<0.05）. The results of HE staining showed that the synovial tissue of the rats in control group was normal， the cells arranged orderly， without edema， hyperemia and proliferation， and had no inflammatory cell infiltration； the synovial tissue of the rats in model group had serious edema and hyperemia，the synovial cells were disordered and proliferaed， and there was a large number of inflammatory cell infiltration； the hyperemia and edema of the synovial tissue of the rats in low dose of diosgenin group were alleviated， and the cell proliferation was slightly reduced； in positive drug group and high dose of diosgenin group， the synovial tissue was slightly edema and congestive， the cells were regularly arranged，and there was small amount of inflammatory cell infiltration and rare proliferation. Compared with control group， the expression levels of TLR2， MyD88 and p-NF-κB p65 proteins in synovial tissue of the rats in model group， positive drug group， low dose of diosgenin group and high dose of diosgenin group were significantly increased （P<0.05）；compared with model group， the expression levels of TLR2， MyD88 and p-NF-κB p65 proteins in synovial tissue of the rats in positive drug group， low dose of diosgenin group and high dose of diosgenin group were significantly increased （P<0.05）；compared with positive group， the expression levels of TLR2， MyD88 and p-NF-κB p65 proteins in synovial tissue of the rats in low and high doses of diosgenin groups were significantly increased （P<0.05）； compared with low dose of diosgenin group， the expression levels of TLR2， MyD88 and p-NF-κB p65 proteins in synovial tissue of the rats in high dose of diosgenin group were significantly decreased（P<0.05）.

Conclusion

Diosgenin may improve the symptoms of synovitis rats by inhibiting the TLR2-NF- κB signaling pathway.

Key words: diosgenin, synovitis, Toll-like receptor 2, nuclear factor-κB, inflammatory cytokines

CLC Number:

R285.5

Haojie WU,Minghui ZHANG,Chengzhi HONG. Improvement effect of diosgenin on symptoms of synovitis rats and its reglatory effect on TLR2-NF-κB signaling pathway and mechanism[J].Journal of Jilin University(Medicine Edition), 2021, 47(4): 943-950.

Figures/Tables 6

Tab. 1

Tab. 2

Tab.3

Fig. 1

Tab.4

Fig. 2

References 20

1	ZHOU L， GU X J. Correlation of ultrasonography synovitis with disease activity and clinical response to etanercept treatment in juvenile idiopathic arthritis patients［J］. Braz J Med Biol Res， 2019， 52（12）： e8565.
2	CHEN H， PAN T， LIU P W， et al. Baihu Jia Guizhi decoction improves rheumatoid arthritis inflammation by regulating succinate/SUCNR1 metabolic signaling pathway［J］. Evid Based Complement Alternat Med， 2019， 2019： 3258572.
3	郭亚春，赵晓菲，黄群，等. 薯蓣皂苷元体外作用对胶原诱导性关节炎小鼠Th1/Th2型细胞因子的影响［J］. 中国老年学杂志， 2018， 38（13）： 3193-3196.
4	CAO C X， WU F， NIU X Y， et al. Cadherin-11 cooperates with inflammatory factors to promote the migration and invasion of fibroblast-like synoviocytes in pigmented villonodular synovitis［J］. Theranostics， 2020， 10（23）： 10573-10588.
5	陈玲，任辉，许蓉，等. 竹节参地上部分总苷成分对大鼠佐剂性关节炎的治疗作用［J］. 中华实验外科杂志， 2019， 36（3）：516-519.
6	郭江燕，姜姝姝，高宇晨，等. 麦粒灸对佐剂型关节炎大鼠屈关节疼痛实验（缩腿嘶鸣）评分及足容积肿胀度的影响［J］. 中医药导报， 2015， 21（11）： 7-9.
7	TREUTLEIN C， BÄUERLE T， NAGEL A M， et al. Comprehensive assessment of knee joint synovitis at 7 T MRI using contrast-enhanced and non-enhanced sequences［J］.BMC Musculoskelet Disord，2020，21（1）：116.
8	ISHIBASHI K， SASAKI E， OTA S， et al. Detection of synovitis in early knee osteoarthritis by MRI and serum biomarkers in Japanese general population［J］. Sci Rep， 2020， 10（1）： 12310.
9	郭亚春，高亚贤，宋鸿儒，等. 薯蓣皂苷元体外对大鼠滑膜RSC-364细胞株STAT3及VEGF影响的研究［J］. 现代预防医学， 2016， 43（1）：128-130，150.
10	王文云，李杰，马铁梁，等. 薯蓣皂苷元对脂多糖诱导的BV2小胶质细胞极化状态的影响［J］. 世界中西医结合杂志， 2018， 13（1）：39-43.
11	LI R， ONG S L， TRAN L M， et al. Chronic IL-1β-induced inflammation regulates epithelial-to-mesenchymal transition memory phenotypes via epigenetic modifications in non-small cell lung cancer［J］. Sci Rep， 2020， 10（1）： 377.
12	UDOMSINPRASERT W， JINAWATH A， TEERAWATTANAPONG N， et al. Interleukin-34 overexpression mediated through tumor necrosis factor-alpha reflects severity of synovitis in knee osteoarthritis［J］. Sci Rep， 2020， 10（1）：7987.
13	CHEN Z， LIN C X， SONG B， et al. Spermidine activates RIP1 deubiquitination to inhibit TNF-α-induced NF-κB/p65 signaling pathway in osteoarthritis［J］. Cell Death Dis， 2020， 11（7）： 503.
14	DING S L， PANG Z Y， CHEN X M， et al. Urolithin a attenuates IL-1β-induced inflammatory responses and cartilage degradation via inhibiting the MAPK/NF-κB signaling pathways in rat articular chondrocytes［J］. J Inflamm （Lond）， 2020， 17： 13.
15	TAN Y， TSENG P， WANG D R， et al. Retraction： stiffening-induced high pulsatility flow activates endothelial inflammation via a TLR2/NF-κB pathway［J］. PLoS One， 2019， 14（7）： e0220600.
16	WU H M， ZHAO C C， XIE Q M， et al. TLR2-melatonin feedback loop regulates the activation of NLRP3 inflammasome in murine allergic airway inflammation［J］. Front Immunol， 2020， 11： 172.
17	CAI X Y， ZHU Y， WANG C， et al. Etanercept inhibits B cell differentiation by regulating TNFRII/TRAF2/NF-κB signaling pathway in rheumatoid arthritis［J］. Front Pharmacol， 2020， 11： 676.
18	SONG H P， ZENG M Y， CHEN X J， et al. Antiulcerogenic activity of Li-Zhong decoction on duodenal ulcers induced by indomethacin in rats： involvement of TLR-2/MyD88 signaling pathway［J］. Evid Based Complement Alternat Med， 2020， 2020： 6538156.
19	MITCHELL J P， CARMODY R J. NF-κB and the transcriptional control of inflammation［J］. Int Rev Cell Mol Biol， 2018， 335： 41-84.
20	崔毅佳，王淑梅，金志国. 甲氨蝶呤通过抑制TLR2-NF-κB信号通路减轻类风湿关节炎大鼠滑膜炎的作用研究［J］. 新疆医科大学学报， 2019， 42（2）：211-216.

Group	n	Pain score
Group	n	（t/d） 3	7	14	21
Control	10	0.64±0.11	0.66±0.12	0.61±0.14	0.65±0.13
Model	10	10.62±1.23^*	12.98±1.16^*□	14.78±1.45^*□▲	16.69±1.98^*□▲●
Positive drug	11	7.14±0.75^*△	5.01±1.52^*△□	3.58±0.74^*△□▲	1.78±0.35^{*△□▲●}
Diosgenin Low dose	12	9.42±1.27^*△#	8.39±1.58^*△#□	7.39±0.71^*△#□▲	4.74±0.45^{*△#□▲●}
High dose	12	8.25±1.28^*△#O	6.77±1.54^*△#O□	4.94±0.77^*△#O□▲	2.78±0.29^{*△#O□▲●}
F		140.339	115.942	395.789	542.709
P		<0.01	<0.01	<0.01	<0.01

Group	n	Toe swelling degree
Group	n	（t/d） 3	7	14	21
Control	10	0.03±0.01	0.04±0.01	0.02±0.01	0.03±0.01
Model	10	0.75±0.08^*	0.97±0.09^*□	1.45±0.19^*□▲	1.61±0.09^*□▲●
Positive drug	11	0.41±0.05^*△	0.32±0.04^*△□	0.24±0.03^*△□▲	0.12±0.02^{*△□▲●}
Diosgenin Low dose	12	0.61±0.07^*△#	0.48±0.08^*△#□	0.37±0.04^*△#□▲	0.24±0.05^{*△#□▲●}
High dose	12	0.53±0.08^*△#○	0.37±0.04^*△#○□	0.29±0.05^{*△#○□▲}	0.19±0.03^{*△#○□▲●}
F		182.726	326.404	418.313	1 920.000
P		<0.01	<0.01	<0.01	<0.01

Group	n	TNF-α	IL-1β
Control	10	61.22±7.03	5.26±0.24
Model	10	180.39±10.06^*	10.47±0.95^*
Positive drug	11	139.25±11.02^*△	5.31±0.53^*△
Diosgenin Low dose	12	104.34±8.04^*△#	8.44±0.74^*△#
High dose	12	90.29±9.03^*△#○	6.98±0.85^*△#○
F		259.719	99.563
P		<0.01	<0.01

Group	n	TLR2	MyD88	NF-κB	p-NF-κB p65
Control	10	0.24±0.06	0.17±0.04	0.59±0.12	0.05±0.01
Model	10	0.85±0.09^*	0.69±0.07^*	0.58±0.15	0.45±0.08^*
Positive drug	11	0.33±0.04^*△	0.32±0.08^*△	0.60±0.14	0.11±0.02^*△
Diosgenin Low dose	12	0.49±0.08^*△#	0.48±0.09^*△#	0.61±0.12	0.36±0.05^*△#
High dose	12	0.37±0.05^{*△# ○}	0.39±0.11^*△#○	0.59±0.11	0.19±0.03^*△#○
F		41.877	54.856	0.088	151.553
P		<0.01	<0.01	0.986	<0.01

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Improvement effect of diosgenin on symptoms of synovitis rats and its reglatory effect on TLR2-NF-κB signaling pathway and mechanism

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