ERβ/MIF信号通路介导的巨噬细胞表型调控在草酸钙结石形成中的作用研究

Published studies have shown that estrogen plays an important role in regulating the formation of calcium oxalate stones. While clinical trials found that estrogen replacement therapy can't effectively reduce the incidence of kidney stones in postmenopausal women, and our previous experiments found that estrogen receptors ERβ antagonists can obviously promote the formation of renal calcium oxalate crystals in mice. These evidences showed that ERβ may play an important regulatory role in the process of calcium oxalate stone formation, but its mechanism is unclear. Recent studies have shown that crystal phagocytosis by macrophages (Mψ) has a key role in the process of reduction of the calcium oxalate stone formation. Our preliminary experiments found that ERβ can influence migration inhibitory factor (MIF) release and regulate Mψ of migration and phenotype transformation. These data may shown that ERβ may regulate the ability of migration, polarization and crystal phagocytosis in Mψ via MIF expression, and finally regulate the formation of calcium oxalate stones, but the mechanism is still unclear. Therefore, this project may propose by transwell culture, chromatin immune coprecipitation and dual luciferase assay,protein kinase chip assay, to clarify the ERβ/MIF signal pathway and mechanism in regulation of Mψ to influence the kidney stone formation in vitro cell and vivo whole body ERβ knockout mice model.

已有的研究表明雌激素对草酸钙结石的形成具有重要调节作用，临床试验却发现补充雌激素并不能有效降低绝经女性肾结石的发病率，且我们前期实验发现雌激素受体ERβ拮抗剂可明显促进小鼠肾脏草酸钙晶体的形成，提示ERβ可能在草酸钙结石形成过程中具有重要调控作用，但其作用及机制尚不清楚。最近研究表明M2型巨噬细胞（Mψ）的晶体内吞作用在减少草酸钙结石形成过程中起重要角色，而且我们前期研究发现ERβ可以通过影响Mψ移动抑制因子(MIF)的释放而调控Mψ的募集和表型转化，提示ERβ可能通过MIF调控Mψ的募集、极化和晶体内吞，继而调控草酸钙结石的形成，但ERβ调控MIF的机制仍不清楚。因此，本项目拟通过Transwell共培养、染色质免疫共沉淀、双荧光素酶报告基因、蛋白激酶芯片检测等方法，在细胞和ERβ全身基因敲除小鼠模型中进一步证实并阐明ERβ通过MIF介导的Mψ表型调控对草酸钙结石形成的作用及分子机制。

已有的研究表明雌激素对草酸钙结石的形成具有重要调节作用，临床试验却发现补充雌激素并不能有效降低绝经女性肾结石的发病率。最近研究表明M2型巨噬细胞(Mψ)的晶体内吞作用在减少草酸钙结石形成过程中起重要角色。本项目拟通过体内及体外实验研究肾小管上皮细胞ERβ与巨噬细胞（Mψ）极化作用，从而影响草酸钙结石形成的机制。项目证实了ERβ可以通过影响Mψ移动抑制因子(MI F)的释放而调控Mψ的募集和表型转化，并影响巨噬细胞对草酸钙晶体内吞能力，继而调控草酸钙结石的形成。通过染色质免疫共沉淀、双荧光素酶报告基因机制研究得出ERβ通过调控miR520f-5p调控MIF的表达。最后通过ERβ全身基因敲除小鼠模型验证了ERβ通过影响MIF及巨噬细胞的募集及表型转化，影响草酸钙晶体在肾脏的形成数量。本研究深入揭示肾结石形成的分子机制提供了新的实验依据，有助于为肾结石的预防提供新的干预策略。