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免疫组库(IR)测序显示,电针通过重新平衡T细胞稳态改善膝骨关节炎。

Electroacupuncture Ameliorates Knee Osteoarthritis By Rebalancing T Cell Homeostasis as Revealed By Immune Repertoire (IR) Sequencing.

作者信息

Jia Wenrui, Zhang Yunan, Wang Tianqi, Liu Cunzhi, Tu Jianfeng, Shi Guangxia, Cai LingYi, Yang Jingwen, Huang Guangrui

机构信息

School of Acupuncture-Moxibustion and Tuina, Beijing University of Chinese Medicine, Beijing 102488, China.

School of Life Science, Beijing University of Chinese Medicine, Beijing 102488, China.

出版信息

Comb Chem High Throughput Screen. 2024 Aug 12. doi: 10.2174/0113862073303471240805061026.

Abstract

BACKGROUND

In this study, we used immune repertoire (IR) sequencing technology to profile the diversity of peripheral blood T cell receptors and used transcriptomics to profile the gene expression of peripheral blood neutrophil mRNA in patients with mild-moderate knee osteoarthritis (KOA) before and after electroacupuncture (EA) treatment.

METHODS

An 8-week intervention with EA was performed on 3 subjects with KOA. IR sequencing of complementarity determining region 3 (CDR3) was performed using RNA extracted from peripheral blood T cells of KOA subjects prior to and at the end of the intervention, as well as healthy volunteers (controls) who matched the subjects in sex and age. Neutrophils were extracted from the plasma of healthy individuals, pretreatment patients, and posttreatment patients for further transcriptome sequencing.

RESULTS

The D50, diversity index (DI), and Shannon entropy values of circulatory T-cells were significantly lower in pretreatment KOA patients compared to healthy controls. Posttreatment KOA samples displayed significant decreases in serum proinflammatory factors, IL-8 and IL-18 (P < 0.01), as well as a substantial reduction in serum matrix MMP-3 and MMP-13 (P < 0.01, P < 0.05). Transcriptome analysis revealed that the expression of CXCL2, IRF8, and PEAR1 (P < 0.05) was significantly higher in patients before the treatment than in the healthy population and was significantly down-regulated after the treatment. In contrast, the expression of SMPD3 (P < 0.05) showed the opposite trend.

CONCLUSION

EA may alleviate KOA by rebalancing T-cell homeostasis and improving systemic inflammation. At the same time, EA treatment can significantly enhance TCR diversity, reduce levels of proinflammatory factors, and increase levels of anti-inflammatory factors, thereby achieving therapeutic effects.

摘要

背景

在本研究中,我们使用免疫组库(IR)测序技术分析外周血T细胞受体的多样性,并运用转录组学分析轻中度膝骨关节炎(KOA)患者在电针(EA)治疗前后外周血中性粒细胞mRNA的基因表达情况。

方法

对3例KOA患者进行为期8周的EA干预。在干预前和干预结束时,从KOA患者以及年龄和性别匹配的健康志愿者(对照)的外周血T细胞中提取RNA,进行互补决定区3(CDR3)的IR测序。从健康个体、治疗前患者和治疗后患者的血浆中提取中性粒细胞,进行进一步的转录组测序。

结果

与健康对照相比,KOA患者治疗前循环T细胞的D50、多样性指数(DI)和香农熵值显著降低。KOA患者治疗后血清促炎因子IL-8和IL-18显著降低(P < 0.01),血清基质金属蛋白酶MMP-3和MMP-13也大幅降低(P < 0.01,P < 0.05)。转录组分析显示,治疗前患者中CXCL2、IRF8和PEAR1的表达(P < 0.05)显著高于健康人群,治疗后显著下调。相反,SMPD3的表达(P < 0.05)呈现相反趋势。

结论

EA可能通过重新平衡T细胞稳态和改善全身炎症来缓解KOA。同时,EA治疗可显著提高TCR多样性,降低促炎因子水平,增加抗炎因子水平,从而实现治疗效果。

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