Effect of Total Saponin of Dioscoreae Collettii Rhizoma on Toll-like Receptor/Nuclear Factor-<italic style="font-style: italic">κ</italic>B (TLR/NF-<italic style="font-style: italic">κ</italic>B) Signaling Pathway Induced by Monosodium Urate in THP-1 Cells

Guo-ying LI; Wei-zhi ZHANG; Lu JIANG; Guang-liang CHEN

doi:10.13422/j.cnki.syfjx.20200401

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Effect of Total Saponin of Dioscoreae Collettii Rhizoma on Toll-like Receptor/Nuclear Factor-κB (TLR/NF-κB) Signaling Pathway Induced by Monosodium Urate in THP-1 Cells

Pharmacology | 更新时间：2021-02-09

- Effect of Total Saponin of Dioscoreae Collettii Rhizoma on Toll-like Receptor/Nuclear Factor-κB (TLR/NF-κB) Signaling Pathway Induced by Monosodium Urate in THP-1 Cells
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 26, Issue 5, Pages: 34-41(2020)
- 作者机构：
  
  1.安徽中医药大学　中西医结合学院，合肥　 230012
  2.安徽省中药复方研究重点实验室，合肥　 230012
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20200401
  CLC： R2-0; R22; R285.5
- Received：03 July 2019，
  
  Published Online：04 November 2019，
  
  Published：05 March 2020
- 稿件说明：
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Guo-ying LI, Wei-zhi ZHANG, Lu JIANG, et al. Effect of Total Saponin of Dioscoreae Collettii Rhizoma on Toll-like Receptor/Nuclear Factor-κB (TLR/NF-κB) Signaling Pathway Induced by Monosodium Urate in THP-1 Cells[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(5): 34-41.
DOI：

Guo-ying LI, Wei-zhi ZHANG, Lu JIANG, et al. Effect of Total Saponin of Dioscoreae Collettii Rhizoma on Toll-like Receptor/Nuclear Factor-κB (TLR/NF-κB) Signaling Pathway Induced by Monosodium Urate in THP-1 Cells[J]. Chinese journal of experimental traditional medical formulae, 2020, 26(5): 34-41. DOI： 10.13422/j.cnki.syfjx.20200401.

摘要

目的：

研究萆薢总皂苷(TSD)对尿酸钠(MSU)诱导THP-1细胞Toll样受体/核转录因子-

B(TLR/NF-

B)信号通路的影响，探讨其抗痛风性关节炎的可能作用机制。

方法：

佛波酯(PMA)诱导THP-1细胞分化为巨噬细胞，分为正常组，模型组，TSD低、中、高质量浓度组(1，3，10 mg·L

-1

)和秋水仙碱组(0.2 mg·L

-1

)，除正常组外，其余各组均用400 mg·L

-1

MSU刺激6 h，诱导体外炎症反应。噻唑蓝(MTT)比色法检测TSD对细胞活力影响；酶联免疫吸附测定法(ELISA)检测细胞上清液炎性因子如肿瘤坏死因子-

(TNF-

)和白细胞介素-1

(IL-1

)的分泌水平；蛋白免疫印迹法(Western blot)检测TLR4，髓样分化因子88(MyD88)及NF-

B蛋白表达；实时荧光定量PCR(Real-time PCR)检测TLR4，NF-

B及IL-1

前体(Pro-IL-1

)mRNA表达水平；免疫荧光法检测NF-

B p65的核位移情况。

结果：

0～32 mg·L

-1

TSD对细胞活力基本无影响；与正常组比较，模型组炎性因子TNF-

及IL-1

的分泌水平显著升高(

0.01)，通路关键蛋白(TLR4，MyD88及NF-

B)和基因(TLR4，NF-

B及Pro-IL-1

)的表达显著升高(

0.01)；与模型组比较，1～30 mg·L

-1

TSD可显著下调炎性因子TNF-

及IL-1

的分泌(

0.01)，通路关键蛋白(TLR4，MyD88及NF-

B)和基因(TLR4，NF-

B及Pro-IL-1

)的表达明显降低(

0.05，

0.01)，细胞核内NF-

B p65叠加减少，部分转位至胞浆，抑制NF-

B p65核转位。

结论：

TSD可能通过下调TLR4，NF-

B及Pro-IL-1

mRNA的表达，减少炎性因子TNF-

及IL-1

分泌而发挥抗炎作用。

Abstract

Objective:

To investigate the effect of total saponin of Dioscoreae Collettii Rhizoma (TSD) on Toll-like receptor/nuclear factor-

B (TLR/NF-

B) signaling pathway induced by monosodium urate in THP-1 cells

in order to explore the possible mechanism of anti-gout arthritis.

Method:

Phorbol 12-myristate 13-acetate (PMA)-induced THP-1 cells were differentiated into macrophages

divided into normal group

model group

low

medium and high-concentration TSD groups (1

10 mg·L

-1

) and colchicine group (0.2 mg·L

-1

). Except the normal group

the other groups were stimulated with 400 mg·L

-1

monosodium urate to replicate an inflammation model

in vitro

. Cell viability was measured by methyl thiazolyl tetrazolium (MTT) assay

the levels of inflammatory factors tumor necrosis factor-

(TNF-

) and interleukin-1

(IL-1

) were detected by enzyme-linked immunosorbent assay (ELISA). The protein levels of Toll-like receptor 4 (TLR4)

myeloid differentiation factor 88 (MyD88) and NF-

B were detected by Western blot. The mRNA levels of TLR4

NF-

B and Pro-IL-1

were measured by real-time fluorescence quantitative PCR (Real-time PCR)

and the nuclear shift of NF-

B p65 was detected by immunofluorescence.

Result:

0~32 mg·L

-1

TSD has no effect on cell viability. Compared with the normal group

the secretion levels of inflammatory factors TNF-

and IL-1

in the model group were significantly increased (

0.01)

and the expressions of key proteins (TLR4

MyD88 and NF-

B) and genes (TLR4

NF-

B and Pro-IL-1

) were increased (

0.01). Compared with the model group

1-30 mg·L

-1

TSD significantly down-regulated the secretion of inflammatory factors TNF-

and IL-1

(

0.01)

the expressions of key proteins (TLR4

MyD88 and NF-

B) and genes (TLR4

NF-

B and Pro-IL-1

) were decreased (

0.05

0.01)

and the NF-

B p65 partially trans-located to the cytosol and the superposition in the nucleus were decreased

inhibiting the nuclear translocation of NF-

B p65.

Conclusion:

TSD may exert an anti-inflammatory effect by down-regulating the expressions of TLR4

NF-

B and Pro-IL-1

mRNA and reducing the secretion of inflammatory factors TNF-

and IL-1

关键词

Keywords

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Effect of Total Saponin of Dioscoreae Collettii Rhizoma on Toll-like Receptor/Nuclear Factor-κB (TLR/NF-κB) Signaling Pathway Induced by Monosodium Urate in THP-1 Cells

DOI：10.13422/j.cnki.syfjx.20200401

摘要

Abstract

关键词

Keywords

references