Calcineurin-NFAT信号通路介导Angptl4在CaN抑制剂治疗MCD中的作用机制研究

Minimal Change Disease (MCD) is one of the most common primary glomerular diseases, which can lead to podocyte cytoskeleton derangement, foot process fusion and proteinuria. As one of the angiopoietin like proteins, Angptl4 can lead to proteinuria, hyperlipidemia, whereas, reduction of Angptl4 with glucocorticoid could obviously reduce proteinuria and hyperlipidemia. Recent researches found calcineurin inhibitor CsA and FK506 can cure MCD, giving us a clue that CsA and FK506 may have some relationship with Angptl4. However, there is no evidence showed their relationship. Our previous results showed that both CsA and FK506 could alleviate the expression of Angptl4 in damaged podocyte, besides, they could also inhibit the apoptosis of podocyte. It has been shown CsA was regulated by calcineurin-NFAT signaling pathway in podocyte. So, we speculate that the expression of Angptl4 in podocyte is regulated by calcineurin -NFAT signaling pathway. So far, there is no report about the relationship between Angptl4 and immunosuppressants. To confirm whether CsA and FK506 protect podocyte by inhibiting the expression of Angptl4 will provide us an important clue for the treatment of MCD. We decide to observe the expression of Angptl4 and make sure whether calcineurin-NFAT signaling pathway can influence its expression in both in vitro mouse podocyte and SD rat model. Our results will provide new clue for the treatment of MCD.

微小病变型肾病（MCD）主要表现为足细胞损伤及大量蛋白尿。Angptl4是血管生成素样蛋白家族成员，和肾脏疾病密切相关。减少Angptl4生成可减轻蛋白尿及高脂血症，说明Angptl4是治疗MCD的靶点。最新研究证实钙调磷酸酶（CaN）抑制剂可治疗MCD，而且其可能和Angptl4相关，但两者的关系迄今国内外尚无报道。本课题组前期结果显示CaN抑制剂可抑制足细胞凋亡及Angptl4的表达。环孢素A可通过调节足细胞中钙调磷酸酶-NFAT信号通路来发挥作用，因此我们推测CaN抑制剂也是通过调节这条信号通路以抑制Angptl4的表达。本项目通过基因转染、基因干扰和示踪等多种技术，从细胞和动物两个层次观察CaN抑制剂对足细胞Angptl4的影响及钙调磷酸酶-NFAT信号通路在影响其表达中的作用。本研究将从新的视觉阐明CaN抑制剂治疗MCD的分子机制，为其治疗提供新的靶点。

微小病变型肾病（MCD）主要表现为足细胞损伤及大量蛋白尿。Angptl4是血管生成素样蛋白家族成员，和肾脏疾病密切相关。研究发现减少Angptl4生成可减轻蛋白尿及高脂血症，说明Angptl4是治疗MCD的靶点。最新研究证实钙调磷酸酶（CaN）抑制剂可治疗MCD，而且其作用可能和Angptl4相关，但两者的关系尚不清楚。本项目通过基因转染、基因干扰等多种技术，从细胞和动物两个层次观察CaN抑制剂对足细胞Angptl4的影响及CaN-NFAT信号通路在影响Angptl4表达中的作用。. 本研究首先通过PAN刺激，建立体内、外小鼠足细胞损伤模型，并检测CaN抑制剂CsA和 FK506对足细胞Angptl4表达的影响。结果显示在体内、外实验中，PAN诱导的足细胞Angptl4的表达均可被CsA和 FK506抑制。随后，我们通过小鼠足细胞NFATc1 转染和NFATc1 的RNA 干扰实验检测Angptl4 表达的变化，在NFATc1 转染的同时，我们也通过NFAT的刺激剂Ionomycin作用于足细胞，观察NFATc1 表达部位的改变及Angptl4的表达变化，结果显示Angptl4的表达随着CaN和NFATc1的变化而变化，说明Angptl4的表达受到Calcineurin-NFATc1信号通路的影响。