背景:牛皮癣,慢性免疫介导的皮肤病,具有病理特征,如角质形成细胞的异常分化,真皮-表皮炎症,和血管生成。2,3,5,4'-四羟基二苯乙烯2-O-β-d-葡萄糖苷(2354Glu)是一种天然小分子聚氢二苯乙烯,分离自何首乌。IL-36亚家族的调节导致了新的药理学策略来逆转银屑病样皮炎。
目的:在这里,我们研究了2354Glu的治疗潜力,并阐明了牛皮癣的潜在机制。
方法:2354Glu对IL-36信号传导的影响通过体内银屑病型评估,体外和离体模型。应用咪喹莫特(IMQ)建立银屑病样皮肤炎症小鼠体内模型,通过刺激小鼠原代角质形成细胞建立体外和体外模型,人角质形成细胞(HaCaT)和离体皮肤组织从小鼠背部分离与聚肌苷-聚胞嘧啶酸(聚(I:C)),IMQ,分别为IL-36γ和脂多糖(LPS)。此外,Cl-脒抑制NETs形成以评价NETs在银屑病小鼠模型中的作用。通过蛋白质印迹评估2354Glu对皮肤炎症的影响,H&E,免疫组织化学,免疫荧光,酶联免疫吸附测定和实时定量PCR。
结果:在聚(I:C)刺激的角质形成细胞中,用2354Glu治疗后,IL-36的分泌受到抑制,与TLR3、P2X7R和caspase-1抑制剂的作用相似。在阿尔达拉(咪喹莫特)诱导的小鼠中,2354Glu(100和25mg/kg)通过P2X7R-caspase-1直接靶向角质形成细胞中的IL-36来改善银屑病中的免疫细胞浸润和角化过度。当用2354Glu(25mg/kg)治疗不足以抑制IL-36γ时,NETs通过与角质形成细胞相互作用以对抗牛皮癣样炎症而降低病理特征和IL-36信号传导。
结论:这些结果表明NETs对银屑病样皮炎具有有益作用。2354Glu通过直接靶向IL-36/P2X7R轴和NET形成缓解银屑病,为牛皮癣的治疗提供了一个潜在的候选人。
BACKGROUND: Psoriasis, a chronic immune-mediated skin disease with pathological features such as aberrant differentiation of keratinocytes, dermal-epidermal inflammation, and angiogenesis. 2,3,5,4\'-Tetrahydroxy stilbene 2-Ο-β-d-glucoside (2354Glu) is a natural small molecule polyhydrostilbenes isolated from Polygonum multiglorum Thunb. The regulation of IL-36 subfamily has led to new pharmacologic strategies to reverse psoriasiform dermatitis.
OBJECTIVE: Here we investigated the therapeutic potential of 2354Glu and elucidated the underlying mechanism in psoriasis.
METHODS: The effects of 2354Glu on IL-36 signaling were assessed by psoriasiform in vivo, in vitro and ex vivo model. The in vivo mice model of psoriasis-like skin inflammation was established by applying imiquimod (IMQ), and the in vitro and ex vitro models were established by stimulating mouse primary keratinocyte, human keratinocytes cells (HaCaT) and ex vivo skin tissue isolated from the mice back with Polyinosine-polycytidylic acid (Poly(I:C)), IMQ, IL-36γ and Lipopolysaccharide (LPS) respectively. Moreover, NETs formation was inhibited by Cl-amidine to evaluate the effect of NETs in psoriatic mouse model. The effects of 2354Glu on skin inflammation were assessed by western blot, H&E, immunohistochemistry, immunofluorescence, enzyme-linked immunosorbent assay and real-time quantitative PCR.
RESULTS: In Poly(I:C)-stimulated keratinocytes, the secretion of IL-36 was inhibited after treatment with 2354Glu, similar to the effects of TLR3, P2X7R and caspase-1 inhibitors. In aldara (imiquimod)-induced mice, 2354Glu (100 and 25 mg/kg) improved immune cell infiltration and hyperkeratosis in psoriasis by directly targeting IL-36 in keratinocytes through P2X7R-caspase-1. When treatment with 2354Glu (25 mg/kg) was insufficient to inhibit IL-36γ, NETs reduced pathological features and IL-36 signaling by interacting with keratinocytes to combat psoriasis like inflammation.
CONCLUSIONS: These results indicated that NETs had a beneficial effect on psoriasiform dermatitis. 2354Glu alleviates psoriasis by directly targeting IL-36/P2X7R axis and NET formation, providing a potential candidate for the treatment of psoriasis.