他莫昔芬通过抑制 H19 激活的 VEGFA 转录发挥抗腹膜纤维化作用。
Tamoxifen exerts anti-peritoneal fibrosis effects by inhibiting H19-activated VEGFA transcription.
机构信息
Department of Nephrology, First Affiliated Hospital of Naval Medical University, Shanghai Changhai Hospital, Shanghai, 200433, China.
Department of Nephrology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital, Shanghai, 200433, China.
出版信息
J Transl Med. 2023 Sep 11;21(1):614. doi: 10.1186/s12967-023-04470-3.
BACKGROUND
Peritoneal dialysis (PD) remains limited due to dialysis failure caused by peritoneal fibrosis. Tamoxifen (TAM), an inhibitor of estrogen receptor 1 (ESR1), has been reported to treat fibrosis, but the underlying mechanism remains unknown. In this study, we sought to explore whether tamoxifen played an anti-fibrotic role by affecting transcription factor ESR1.
METHODS
ESR1 expression was detected in the human peritoneum. Mice were daily intraperitoneally injected with 4.25% glucose PD dialysate containing 40 mM methylglyoxal for 2 weeks to establish PD-induced peritoneal fibrosis. Tamoxifen was administrated by daily gavage, at the dose of 10 mg/kg. Chromatin immunoprecipitation (ChIP) and dual-luciferase reporter assay were performed to validate ESR1 bound H19 promoter. Gain-of-function and loss-of-function experiments were performed to investigate the biological roles of H19 on the mesothelial-mesenchymal transition (MMT) of human peritoneal mesothelial cells (HPMCs). Intraperitoneal injection of nanomaterial-wrapped 2'-O-Me-modified small interfering RNA was applied to suppress H19 in the mouse peritoneum. RNA immunoprecipitation and RNA pull-down assays demonstrated binding between H19 and p300. Exfoliated peritoneal cells were obtained from peritoneal dialysis effluent to analyze the correlations between ESR1 (or H19) and peritoneal solute transfer rate (PSTR).
RESULTS
ESR1 was increased significantly in the peritoneum after long-term exposure to PD dialysate. Tamoxifen treatment ameliorated high glucose-induced MMT of HPMCs, improved ultrafiltration rate, and decreased PSTR of mouse peritoneum. Tamoxifen reduced the H19 level by decreasing the ESR1 transcription of H19. Depletion of H19 reversed the pro-fibrotic effect of high glucose while ectopic expression of H19 exacerbated fibrotic pathological changes. Intraperitoneal injection of nanomaterial-wrapped 2'-O-Me-modified siRNAs targeting H19 mitigated PD-related fibrosis in mice. RNA immunoprecipitation (RIP) and RNA pull-down results delineated that H19 activated VEGFA expression by binding p300 to the VEGFA promoter and inducing histone acetylation of the VEGFA promoter. ESR1 and H19 were promising targets to predict peritoneal function.
CONCLUSIONS
High glucose-induced MMT of peritoneal mesothelial cells in peritoneal dialysis via activating ESR1. In peritoneal mesothelial cells, ESR1 transcribed the H19 and H19 binds to transcription cofactor p300 to activate the VEGFA. Targeting ESR1/H19/VEGFA pathway provided new hope for patients undergoing peritoneal dialysis.
背景
由于腹膜纤维化导致透析失败，腹膜透析（PD）仍然受到限制。他莫昔芬（TAM）是一种雌激素受体 1（ESR1）抑制剂，已被报道可用于治疗纤维化，但具体机制尚不清楚。在这项研究中，我们试图通过影响转录因子 ESR1 来探讨他莫昔芬是否具有抗纤维化作用。
方法
检测人腹膜中的 ESR1 表达。通过每日腹腔内注射 4.25%葡萄糖 PD 透析液（含 40 mM 甲基乙二醛）2 周，建立 PD 诱导的腹膜纤维化小鼠模型。他莫昔芬通过每日灌胃给药，剂量为 10 mg/kg。通过染色质免疫沉淀（ChIP）和双荧光素酶报告基因检测验证 ESR1 结合 H19 启动子。通过过表达和敲低实验研究 H19 对人腹膜间皮细胞（HPMCs）间皮-间充质转化（MMT）的生物学作用。通过腹腔内注射纳米材料包裹的 2'-O-Me 修饰的小干扰 RNA 抑制小鼠腹膜中的 H19。RNA 免疫沉淀和 RNA 下拉实验表明 H19 与 p300 结合。从腹膜透析流出液中获得脱落的腹膜细胞，分析 ESR1（或 H19）与腹膜溶质转运率（PSTR）之间的相关性。
结果
长期暴露于 PD 透析液后，腹膜中 ESR1 的表达显著增加。他莫昔芬治疗可改善高糖诱导的 HPMCs MMT，提高超滤率，并降低小鼠腹膜 PSTR。他莫昔芬通过降低 ESR1 对 H19 的转录来降低 H19 水平。高糖条件下，H19 耗竭可逆转促纤维化作用，而过表达 H19 则加剧纤维化病理变化。腹腔内注射纳米材料包裹的 2'-O-Me 修饰靶向 H19 的 siRNA 可减轻小鼠 PD 相关纤维化。RNA 免疫沉淀（RIP）和 RNA 下拉结果表明，H19 通过与 VEGFA 启动子结合并诱导 VEGFA 启动子组蛋白乙酰化来激活 VEGFA 的表达。ESR1 和 H19 是预测腹膜功能的有前途的靶点。
结论
高糖通过激活 ESR1 诱导腹膜透析中腹膜间皮细胞的 MMT。在腹膜间皮细胞中，ESR1 转录 H19，H19 与转录共因子 p300 结合，激活 VEGFA。靶向 ESR1/H19/VEGFA 通路为腹膜透析患者带来了新的希望。
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