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双特异性磷酸酶3和TNF-α对雷公藤多苷片治疗类风湿关节炎的预后作用及其受调节机制
Dual specificity protein phosphatase 3 and tumor necrosis factor-α as prognostic biomarkers and their regulation by tripterygium glycosides in rheumatoid arthritis
- 第三军医大学学报 2019年41卷第1期 页码：77-84
- 作者机构：
237006 安徽 六安，六安市人民医院风湿免疫科
- 作者简介：
[ 李兴锐，E-mail：13956113863@163.com LI Xingrui, E-mail:13956113863@163.com
- 基金信息：安徽省卫生计生委全科医学临床科研课题(2016QK028)
DOI：10.16016/j.1000-5404.201806039
中图分类号： R285.6;R392.5;R593.22收稿：2018-06-07，
修回：2018-08-30，
纸质出版：2019-01-15
- 稿件说明：
移动端阅览
李兴锐, 谭悦, 刘童, 等. 双特异性磷酸酶3和TNF-α对雷公藤多苷片治疗类风湿关节炎的预后作用及其受调节机制[J]. 第三军医大学学报, 2019,41(1):77-84.
Xingrui LI, Yue TAN, Tong LIU, et al. Dual specificity protein phosphatase 3 and tumor necrosis factor-α as prognostic biomarkers and their regulation by tripterygium glycosides in rheumatoid arthritis[J]. Journal of Third Military Medical University, 2019, 41(1): 77-84.
李兴锐, 谭悦, 刘童, 等. 双特异性磷酸酶3和TNF-α对雷公藤多苷片治疗类风湿关节炎的预后作用及其受调节机制[J]. 第三军医大学学报, 2019,41(1):77-84. DOI： 10.16016/j.1000-5404.201806039.
Xingrui LI, Yue TAN, Tong LIU, et al. Dual specificity protein phosphatase 3 and tumor necrosis factor-α as prognostic biomarkers and their regulation by tripterygium glycosides in rheumatoid arthritis[J]. Journal of Third Military Medical University, 2019, 41(1): 77-84. DOI： 10.16016/j.1000-5404.201806039.
摘要
目的
2
探讨双特异性磷酸酶3（dual specificity protein phosphatase 3，DUSP3）和TNF-α作为雷公藤多苷（tripterygium glycosides，TG）片治疗类风湿关节炎（rheumatoid arthritis，RA）预后生物标记物的可行性和调节机制。
方法
2
选取本院于2016年3月至2018年1月收治的67例RA患者及67例健康志愿者，RT-PCR检测治疗前后外周血单个核细胞（peripheral blood mononuclear cells，PBMC）中DUSP3和TNF-α表达，并进行ROC分析。以TG处理脂多糖刺激的U937细胞，模拟TG片治疗RA，通过RT-PCR、Western blot和免疫共沉淀实验检测DUSP3和TNF-α之间的关系，及其上下游通路。
结果
2
与健康志愿者相比，RA患者（用药前）PBMC中DUSP3表达降低而TNF-α表达升高（
P
<
0.05）。TNF-α和DUSP3进行ROC分析的AUC分别为0.948和0.908。治疗后RA患者DAS28，ESR和CRP值下降，DUSP3和TNF-α表达分别升高和降低（
P
<
0.05）。TG片用药响应和不响应（骨侵蚀指标）的RA患者的DUSP3和TNF-α表达水平差异显著（
P
<
0.05）。以用药后DAS28表达变化区分RA患者是否响应TG片治疗，ROC分析显示DUSP3和TNF-α的AUC分别为0.821和0.747。在LPS刺激的U937细胞中，改变DUSP3或TNF-α表达不相互影响表达。Western blot结果显示NF-κB介导TG对TNF-α调节。ChIP结果证实TG抑制了HDAC1与DUSP3启动子区相互作用。Western blot结果表明TG给药抑〖JP+1〗制了LPS诱导的HDAC1上调和DUSP3下调。
结论
2
TG片对RA有较好的治疗效果。其中，DUSP3和TNF-α可以作为TG片治疗RA的预后因子。TG给药降低了TNF-α的表达，并解除了对DUSP3的抑制。
Abstract
Objective
2
To investigate the prognostic value of dual specificity protein phosphatase 3(DUSP3) and tumor necrosis factor-α(TNF-α) and how tripterygium glycosides(TG) regulate DUSP3 and TNF-α expression in patients with rheumatoid arthritis(RA).
Methods
2
A total of 67 patients with RA receiving treatment with TG tablets in our hospital between March
2016 and January
2018 and 67 healthy volunteers were enrolled in this study. The expression of DUSP3 and TNF-α in the peripheral blood mononuclear cells(PBMCs) of the participants was detected using real-time PCR(RT-PCR)
and the results were analyzed using the receiver operating curve(ROC). In U937 cells stimulated with lipopolysaccharide(LPS)
the effects of TG on the expressions of DUSP3
TNF-α and their upstream and downstream pathways were investigated using RT-PCR
Western blotting and chromatin immunoprecipitation(ChIP).
Results
2
Compared with the healthy volunteers
the RA patients showed significantly reduced DUSP3 expression and increased TNF-α expression in the PBMCs(
P
<
0.05)
whose areas under the curve(AUC) were 0.948 and 0.908
respectively. Treatment with TG tab
lets resulted in significantly decreased 28-joint Disease Activity score(DAS28)
erythrocyte sedimentation rate(ESR) and C-reactive protein(CRP)
increased expression of DUSP3 and decreased TNF-α expression in the RA patients(
P
<
0.05). Significant differences were found in the expression of DUSP3 and TNF-α between the RA patients who responded to the treatment and the non-responding patients(with bone erosion
P
<
0.05). ROC analysis showed that the AUC of DUSP3 and TNF-α was 0.821 and 0.747
respectively
for predicting the patients' response to TG treatment(defined by the changes in DAS28). In LPS-stimulated U937 cells
the changes in DUSP3 or TNF-α expression did not affect the expression of one another. The rescue experiments demonstrated that nuclear factor-κB(NF-κB) mediated the inhibitory effect of TG on TNF-α expression. ChIP experiments confirmed that TG suppressed the conjugation of HDAC1 with DUSP3 promoter; Western blotting showed that TG inhibited LPS-induced up-regulation of HDAC1 and down-regulation of DUSP3.
Conclusion
2
TG tablet produces good therapeutic effect on RA by reducing the expression of TNF-α and relieving the inhibition on DUSP3. DUSP3 and TNF-α can serve as prognostic biomarkers for RA patients receiving treatment with TG tablets.
关键词
Keywords
references
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