The prevalence of lupus nephritis (LN) remains high despite various emerging monoclonal antibodies against with targeting systemic lupus erythematosus (SLE). Renal fibrosis is the main feature of late stage LN, and novel therapeutic agents are still needed. We previously reported that microRNA (miR)-150 increases in renal biopsies of American LN patients and that miR-150 agonist promotes fibrosis in cultured kidney cells. Presently, we aim to verify whether locked nucleic acid (LNA)-anti-miR-150 can ameliorate LN in mice and to investigate its corresponding mechanisms. We first observed natural history and renal miR-150 expression in female Fcgr2b−/− mice of a spontaneously developed LN model. We then verified miR-150 renal absorption and determined the dose of the suppressed miR-150 by subcutaneous injection of LNA-anti-miR-150 (2 and 4 mg/kg). Thirdly, we investigated the therapeutic effects of LNA-anti-miR-150 (2 mg/kg for 8 weeks) on LN mice and the corresponding mechanisms by studying fibrosis-related genes, cytokines, and kidney resident macrophages. Lastly, we detected the expression of renal miR-150 and the mechanism-associated factors in renal biopsies from new onset untreated LN patients. Fcgr2b−/− mice developed SLE indicated by positive serum autoantibodies at age 19 weeks and LN demonstrated by proteinuria at age 32 weeks. Renal miR-150 was overexpressed in LN mice compared to wild type mice. FAM-labeled LNA-anti-miR-150 was absorbed by both glomeruli and renal tubules. LNA-anti-miR-150 suppressed the elevated renal miR-150 levels in LN mice compared to the scrambled LNA without systemic toxicity. Meanwhile, serum double strand-DNA antibody, proteinuria, and kidney injury were ameliorated. Importantly, the elevated renal pro-fibrotic genes (transforming growth factor-β1, α-smooth muscle antibody, and fibronectin) and decreased anti-fibrotic gene suppressor of cytokine signal 1 were both reversed. Renal pro-inflammatory cytokines (interferon-γ, interleukin-6, and tumor necrosis factor-α) and macrophages were also decreased. In addition, the changes of renal miR-150 and associated proteins shown in LN mice were also seen in human subjects. LNA-anti-miR-150 may be a promising novel therapeutic agent for LN in addition to the current emerging monoclonal antibodies, and its renal protective mechanism may be mediated by anti-fibrosis and anti-inflammation as well as reduction of the infiltrated kidney resident macrophages.

中文翻译：

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LNA-抗miR-150通过抑制肾纤维化和巨噬细胞浸润改善了狼疮性肾炎的肾脏损伤。

尽管出现了针对靶向系统性红斑狼疮（SLE）的各种新兴单克隆抗体，但狼疮性肾炎（LN）的患病率仍然很高。肾纤维化是晚期LN的主要特征，仍然需要新型治疗剂。我们以前曾报道过，美国LN患者的肾脏活检中microRNA（miR）-150升高，而miR-150激动剂可促进培养的肾细胞中的纤维化。目前，我们旨在验证锁核酸（LNA）-抗miR-150是否可以改善小鼠的LN，并研究其相应的机制。我们首先观察了自然发展史和肾脏miR-150在自发形成的LN模型的雌性Fcgr2b-/-小鼠中的表达。然后，我们通过皮下注射LNA-抗miR-150（2和4 mg / kg）验证了miR-150的肾脏吸收并确定了抑制的miR-150的剂量。第三，我们通过研究与纤维化相关的基因，细胞因子和肾驻留巨噬细胞，研究了LNA-抗miR-150（2 mg / kg，持续8周）对LN小鼠的治疗作用以及相应的机制。最后，我们在未经治疗的新发LN患者的肾脏活检中检测了肾脏miR-150的表达及其机制相关因素。Fcgr2b-/-小鼠在19周龄时出现血清自身抗体阳性表示SLE，而在32周龄时通过蛋白尿表现出LN。与野生型小鼠相比，LN小鼠中的肾miR-150过表达。FAM标记的LNA-抗miR-150被肾小球和肾小管吸收。与没有全身毒性的混乱LNA相比，LNA-抗miR-150抑制了LN小鼠中升高的肾脏miR-150水平。同时，血清双链DNA抗体，蛋白尿，和肾脏损伤得到改善。重要的是，升高的肾促纤维化基因（转化生长因子-β1，α-平滑肌抗体和纤连蛋白）和细胞因子信号1的抗纤维化基因抑制子均被逆转。肾促炎细胞因子（干扰素-γ，白介素-6和肿瘤坏死因子-α）和巨噬细胞也减少了。此外，在人类受试者中也观察到了LN小鼠中显示的肾脏miR-150和相关蛋白的变化。除目前新兴的单克隆抗体外，LNA-miR-150可能是一种有前途的新型LN治疗药物，其肾脏保护机制可能由抗纤维化和抗炎作用以及减少肾浸润的居民介导。巨噬细胞。升高的肾促纤维化基因（转化生长因子-β1，α-平滑肌抗体和纤连蛋白）和细胞因子信号1的抗纤维化基因抑制子均被逆转。肾促炎细胞因子（干扰素-γ，白介素-6和肿瘤坏死因子-α）和巨噬细胞也减少了。此外，在人类受试者中也观察到了LN小鼠中显示的肾脏miR-150和相关蛋白的变化。除目前新兴的单克隆抗体外，LNA-抗miR-150可能是一种有前途的新型LN治疗药物，其肾脏保护机制可能由抗纤维化和抗炎作用以及减少的肾脏浸润介导巨噬细胞。升高的肾促纤维化基因（转化生长因子-β1，α-平滑肌抗体和纤连蛋白）和细胞因子信号1的抗纤维化基因抑制子均被逆转。肾促炎细胞因子（干扰素-γ，白介素-6和肿瘤坏死因子-α）和巨噬细胞也减少了。此外，在人类受试者中也观察到了LN小鼠中显示的肾脏miR-150和相关蛋白的变化。除目前新兴的单克隆抗体外，LNA-miR-150可能是一种有前途的新型LN治疗药物，其肾脏保护机制可能由抗纤维化和抗炎作用以及减少肾浸润的居民介导。巨噬细胞。和纤连蛋白）和细胞因子信号1降低的抗纤维化基因抑制剂均被逆转。肾促炎细胞因子（干扰素-γ，白介素-6和肿瘤坏死因子-α）和巨噬细胞也减少了。此外，在人类受试者中也观察到了LN小鼠中显示的肾脏miR-150和相关蛋白的变化。除目前新兴的单克隆抗体外，LNA-miR-150可能是一种有前途的新型LN治疗药物，其肾脏保护机制可能由抗纤维化和抗炎作用以及减少肾浸润的居民介导。巨噬细胞。和纤连蛋白）和细胞因子信号1降低的抗纤维化基因抑制剂均被逆转。肾促炎细胞因子（干扰素-γ，白介素-6和肿瘤坏死因子-α）和巨噬细胞也减少了。此外，在人类受试者中也观察到了LN小鼠中显示的肾脏miR-150和相关蛋白的变化。除目前新兴的单克隆抗体外，LNA-miR-150可能是一种有前途的新型LN治疗药物，其肾脏保护机制可能由抗纤维化和抗炎作用以及减少肾浸润的居民介导。巨噬细胞。在人类受试者中也观察到了LN小鼠中显示的肾miR-150和相关蛋白的变化。除目前新兴的单克隆抗体外，LNA-miR-150可能是一种有前途的新型LN治疗药物，其肾脏保护机制可能由抗纤维化和抗炎作用以及减少肾浸润的居民介导。巨噬细胞。在人类受试者中也观察到了LN小鼠中显示的肾miR-150和相关蛋白的变化。除目前新兴的单克隆抗体外，LNA-miR-150可能是一种有前途的新型LN治疗药物，其肾脏保护机制可能由抗纤维化和抗炎作用以及减少肾浸润的居民介导。巨噬细胞。