羟氯喹(HCQ)是杂环芳香化合物喹啉的衍生物,已用于治疗自身免疫性疾病。本研究的主要目的是研究HCQ对实验性自身免疫性肝炎(AIH)的治疗作用和炎症免疫分子机制。HCQ治疗可改善肝脏病理损伤,炎性浸润,同时促进S-100抗原诱导的AIH小鼠调节性T细胞(Treg)和下调CD8+T细胞分化。体外,HCQ还抑制促炎细胞因子(IFN-γ,TNF-α,和IL-12)分泌,促进抗炎细胞因子(TGF-β1)的分泌。HCQ主要损害T细胞脂质代谢,但不促进糖酵解,以促进Treg的分化和功能。机械上,HCQ下调GRK2在T细胞中的膜转位,抑制GRK2-PI3K相互作用以减少PI3K对膜的募集,然后抑制PI3K-AKT-mTOR信号的磷酸化。用帕罗西汀预处理T细胞,GRK2抑制剂,干扰HCQ对T细胞的影响。HCQ还通过740Y-P(PI3K激动剂)逆转PI3K-AKT轴的活化。同时,HCQ抑制PI3K-AKT-mTOR,JAK2-STAT3-SOCS3和增加AIH小鼠肝脏和T细胞中的AMPK信号。总之,HCQ对AIH和伴随的肝损伤表现出特定和有效的治疗作用,这归因于HCQ作用于GRK2易位,抑制T淋巴细胞中代谢相关的PI3K-AKT和炎症相关的JAK2-STAT3信号,从而调节T细胞的脂质代谢功能来调节Treg的分化和功能。
Hydroxychloroquine (HCQ) is derivative of the heterocyclic aromatic compound quinoline, which has been used for the treatment of autoimmune diseases. The central purpose of this study was to investigate therapeutic effects and inflammatory immunological molecular mechanism of HCQ in experimental autoimmune hepatitis (AIH). Treatment with HCQ ameliorated hepatic pathologic damage, inflammatory infiltration, while promoted regulatory T cell (Treg) and down-regulated CD8+T cell differentiation in AIH mice induced by S-100 antigen. In vitro, HCQ also suppressed pro-inflammatory cytokine (IFN-γ, TNF-α, and IL-12) secretion, promoted anti-inflammatory cytokine (TGF-β1) secretion. HCQ mainly impaired T cell lipid metabolism but not glycolysis to promote Treg differentiation and function. Mechanistically, HCQ down-regulated GRK2 membrane translocation in T cells, inhibited GRK2-PI3K interaction to reduce the PI3K recruiting to the membrane, followed by suppressing the phosphorylation of PI3K-AKT-mTOR signal. Pretreating T cells with paroxetine, a GRK2 inhibitor, disturbed HCQ effect to T cells. HCQ also reversed the activation of the PI3K-AKT axis by 740 Y-P (PI3K agonist). Meanwhile, HCQ inhibited the PI3K-AKT-mTOR, JAK2-STAT3-SOCS3 and increased the AMPK signals in the liver and T cells of AIH mice. In conclusion, HCQ exhibited specific and potent therapeutic effects on AIH and attendant liver injury, which was attributed to HCQ acted on GRK2 translocation, inhibited metabolism-related PI3K-AKT and inflammation-related JAK2-STAT3 signal in T lymphocytes, thereby modulating lipid metabolism of T cell function to regulate Treg differentiation and function.