Laboratório de Inflamação, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Departamento de Ciências da Natureza, Instituto de Aplicação Fernando Rodrigues da Silveira, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brazil.
PLoS Pathog. 2024 Aug 22;20(8):e1011812. doi: 10.1371/journal.ppat.1011812. eCollection 2024 Aug.
Identifying new molecular therapies targeted at the severe hepatic fibrosis associated with the granulomatous immune response to Schistosoma mansoni infection is essential to reduce fibrosis-related morbidity/mortality in schistosomiasis. In vitro cell activation studies suggested the lipid molecule prostaglandin D2 (PGD2) as a potential pro-fibrotic candidate in schistosomal context, although corroboratory in vivo evidence is still lacking. Here, to investigate the role of PGD2 and its cognate receptor DP2 in vivo, impairment of PGD2 synthesis by HQL-79 (an inhibitor of the H-PGD synthase) or DP2 receptor inhibition by CAY10471 (a selective DP2 antagonist) were used against the fibrotic response of hepatic eosinophilic granulomas of S. mansoni infection in mice. Although studies have postulated PGD2 as a fibrogenic molecule, HQL-79 and CAY10471 amplified, rather than attenuated, the fibrotic response within schistosome hepatic granulomas. Both pharmacological strategies increased hepatic deposition of collagen fibers - an unexpected outcome accompanied by further elevation of hepatic levels of the pro-fibrotic cytokines TGF-β and IL-13 in infected animals. In contrast, infection-induced enhanced LTC4 synthesis in the schistosomal liver was reduced after HQL-79 and CAY10471 treatments, and therefore, inversely correlated with collagen production in granulomatous livers. Like PGD2-directed maneuvers, antagonism of cysteinyl leukotriene receptors CysLT1 by MK571 also promoted enhancement of TGF-β and IL-13, indicating a key down-regulatory role for endogenous LTC4 in schistosomiasis-induced liver fibrosis. An ample body of data supports the role of S. mansoni-driven DP2-mediated activation of eosinophils as the source of LTC4 during infection, including: (i) HQL-79 and CAY10471 impaired systemic eosinophilia, drastically decreasing eosinophils within peritoneum and hepatic granulomas of infected animals in parallel to a reduction in cysteinyl leukotrienes levels; (ii) peritoneal eosinophils were identified as the only cells producing LTC4 in PGD2-mediated S. mansoni-induced infection; (iii) the magnitude of hepatic granulomatous eosinophilia positively correlates with S. mansoni-elicited hepatic content of cysteinyl leukotrienes, and (iv) isolated eosinophils from S. mansoni-induced hepatic granuloma synthesize LTC4 in vitro in a PGD2/DP2 dependent manner. So, our findings uncover that granulomatous stellate cells-derived PGD2 by activating DP2 receptors on eosinophils does stimulate production of anti-fibrogenic cysLTs, which endogenously down-regulates the hepatic fibrogenic process of S. mansoni granulomatous reaction - an in vivo protective function which demands caution in the future therapeutic attempts in targeting PGD2/DP2 in schistosomiasis.
鉴定针对曼氏血吸虫感染引起的严重肝纤维化的新型分子治疗方法对于降低血吸虫病相关的纤维化发病率/死亡率至关重要。体外细胞激活研究表明,脂质分子前列腺素 D2(PGD2)是血吸虫病背景下潜在的促纤维化候选物,尽管目前仍缺乏确证性的体内证据。在这里,为了研究 PGD2 及其同源受体 DP2 在体内的作用,使用 HQL-79(H-PGD 合酶抑制剂)抑制 PGD2 合成或 CAY10471(选择性 DP2 拮抗剂)抑制 DP2 受体,以对抗曼氏血吸虫感染小鼠肝嗜酸性肉芽肿的纤维化反应。尽管有研究假设 PGD2 是一种纤维生成分子,但 HQL-79 和 CAY10471 增强而非减弱了血吸虫肝肉芽肿内的纤维化反应。这两种药理策略都增加了胶原纤维在肝脏中的沉积 - 这是一种出乎意料的结果,伴随着感染动物肝内促纤维化细胞因子 TGF-β 和 IL-13 的进一步升高。相比之下,在 HQL-79 和 CAY10471 治疗后,感染诱导的肝内 LTC4 合成增强减少,因此与肉芽肿性肝脏中的胶原产生呈负相关。与 PGD2 靶向操作一样,MK571 对半胱氨酰白三烯受体 CysLT1 的拮抗作用也促进了 TGF-β 和 IL-13 的增强,表明内源性 LTC4 在血吸虫病诱导的肝纤维化中具有关键的下调作用。大量数据支持 S. mansoni 驱动的 DP2 介导的嗜酸性粒细胞激活作为感染期间 LTC4 的来源的作用,包括:(i)HQL-79 和 CAY10471 损害了全身嗜酸性粒细胞增多症,在感染动物的腹膜和肝肉芽肿中急剧减少了嗜酸性粒细胞,同时降低了半胱氨酰白三烯的水平;(ii)腹膜嗜酸性粒细胞被鉴定为在 PGD2 介导的 S. mansoni 诱导感染中产生 LTC4 的唯一细胞;(iii)肝肉芽肿性嗜酸性粒细胞的数量与 S. mansoni 引起的肝半胱氨酰白三烯含量呈正相关;(iv)从 S. mansoni 诱导的肝肉芽肿中分离出的嗜酸性粒细胞在体外以 PGD2/DP2 依赖的方式合成 LTC4。因此,我们的研究结果表明,通过激活嗜酸性粒细胞上的 DP2 受体,由肉芽肿性星状细胞衍生的 PGD2 确实刺激了抗纤维化的 cysLTs 的产生,这内源性地下调了 S. mansoni 肉芽肿反应的肝纤维化过程 - 这是一种体内保护功能,在未来针对血吸虫病中 PGD2/DP2 的治疗尝试中需要谨慎。