目的:本研究旨在从ceRNA的角度探讨通过抑制Treg促进系统性红斑狼疮(SLE)活性的机制。
方法:qRT-PCR检测临床SLE患者样本中circETS1、miR-1205和FoxP3的表达。在CD4+T细胞中进行circETS1和miR-1205的过表达,以及miR-1205和FoxP3的敲低,同时检测到辅助性T细胞17(Th17)和调节性T细胞(Treg)的增殖。进行了脱氮实验以验证circETS1/miR-1205/Foxp3mRNA轴调节CD4T细胞分化的分子机制。在体内实验中,miR-1205在SLE小鼠中的表达被干预,肾功能,炎症因子,并测定血清补体。此外,流式细胞术检测Treg/Th17细胞比例。
结果:在SLE患者中,发现Treg细胞减少,而Th17细胞增加。转染circETS1过表达导致CD4+T细胞分化成Treg细胞,导致Th17/Treg比例失衡。转染miR-1205模拟物和si-FoxP3可以逆转circETS1过表达的作用。此外,抑制miR-1205的表达对SLE小鼠有治疗作用。
结论:circETS1通过miR-1205/FoxP3轴抑制Treg,从而促进SLE活动,可能成为SLE治疗的新靶点。
OBJECTIVE: This study aimed to explore the mechanism by which systemic lupus erythematosus (SLE) activity is promoted through Treg inhibition from the perspective of ceRNA.
METHODS: qRT-PCR was used to detect the expressions of circETS1, miR-1205, and FoxP3 in clinical SLE patient samples. Overexpression of circETS1and miR-1205, along with knockdown of miR-1205 and FoxP3 were conducted in CD4+ T cells, while the proliferation of helper T cell 17 (Th17) and regulatory T cell (Treg) was detected. Arescue assay was performed to verify the molecular mechanism of circETS1/miR-1205/Foxp3 mRNA axis in regulating CD4+ T cell differentiation. In the in vivo experiment, the expression of miR-1205 in SLE mice was intervened, and renal function, inflammatory factors, and serum complement were measured. Additionally, Treg/Th17 cell ratio was detected by flow cytometry.
RESULTS: In SLE patients, Treg cells were found to decrease, while Th17 cells increased. Transfection with circETS1 overexpression led to CD4+ T cells differentiating into Treg cells, causing an imbalance in the Th17/Treg ratio. Transfection of miR-1205 mimic and si-FoxP3 could reverse the effect of circETS1 overexpression. Moreover, inhibiting the expression of miR-1205 showed therapeutic effects on SLE mice.
CONCLUSIONS: circETS1 inhibits Treg via the miR-1205/FoxP3 axis, thereby promoting SLE activity, which may become a new target for SLE treatment.