ABSTRACT

In clinical practice, we found that microRNA (miR)-146a-5p is significantly up-regulated in peripheral blood mononuclear cells (PBMCs) of primary sjögren’s syndrome (pSS) patients. In vitro experiments confirmed that miR-146a-5p promotes T helper 17 (Th17) cell differentiation, but the specific mechanism is still unknown. To solve this problem, 20 pSS patients and 20 healthy subjects were enrolled in this study and PBMCs were isolated from their blood. The expression of the membrane IL-23 R (mIL-23 R) in PBMCs was determined. CD3⁺ T cells were also isolated and used to further analyze the relationship between the ectodomain shedding of mIL-23 R and a disintegrin and metalloprotease 17 (ADAM17). Finally, miR-146a-5p inhibitor and mimics were transfected into PBMCs to evaluate the relationship between ADAM17 and mIL-23 R, and explore the role of mIL-23 R and ADAM17 in Th17 cell differentiation. Our results revealed a significantly increased expression of miR-146a-5p in PBMCs from pSS patients and significantly increased percentage of Th17 cells compared to PBMCs from healthy controls. Under polarization culture conditions, pSS patient-derived PBMCs can more easily differentiate into Th17 cells, which was, to a great extent, attributable to the increased expression of mIL-23 R. Moreover, ADAM17, an ectodomain sheddase of mIL-23 R, was targeted and negatively regulated by miR-146a-5p, which reduced the ectodomain shedding of mIL-23 R. Overall, our results suggested that miR-146a-5p could promote Th17 cell differentiation through targeting and negatively regulating ADAM17. Thus, these results might offer a new approach in the treatment of pSS.

摘要

在临床实践中，我们发现 microRNA (miR)-146a-5p 在原发性干燥综合征 (pSS) 患者的外周血单个核细胞 (PBMC) 中显着上调。体外实验证实miR-146a-5p促进T辅助17（Th17）细胞分化，但具体机制尚不清楚。为了解决这个问题，本研究招募了 20 名 pSS 患者和 20 名健康受试者，并从他们的血液中分离出 PBMC。测定了 PBMC 中膜 IL-23 R (mIL-23 R) 的表达。CD3 ⁺还分离了 T 细胞并用于进一步分析 mIL-23 R 的胞外域脱落与去整合素和金属蛋白酶 17 (ADAM17) 之间的关系。最后，将miR-146a-5p抑制剂和模拟物转染到PBMC中，评估ADAM17和mIL-23 R之间的关系，并探讨mIL-23 R和ADAM17在Th17细胞分化中的作用。我们的结果显示，与健康对照的 PBMC 相比，pSS 患者的 PBMC 中 miR-146a-5p 的表达显着增加，Th17 细胞的百分比显着增加。在极化培养条件下，pSS 患者来源的 PBMC 更容易分化为 Th17 细胞，这在很大程度上归因于 mIL-23 R 的表达增加。此外，mIL-23 R 的胞外域脱落酶 ADAM17，被 miR-146a-5p 靶向和负调控，这减少了 mIL-23 R 的胞外域脱落。总的来说，我们的结果表明 miR-146a-5p 可以通过靶向和负调控 ADAM17 促进 Th17 细胞分化。因此，这些结果可能为治疗 pSS 提供一种新方法。