Fractalkine deficiency attenuates LPS-induced acute kidney injury and podocyte apoptosis by targeting the PI3K/Akt signal pathway
Fractalkine deficiency attenuates LPS-induced acute kidney injury and podocyte apoptosis by targeting the PI3K/Akt signal pathway
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Fractalkine 缺陷通过靶向 PI3K/Akt 信号通路减轻 LPS 诱导的急性肾损伤和足细胞凋亡
DOI：
10.1007/s10157-022-02218-9
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发表时间：
2022-04-08
影响因子：
2.3
通讯作者：
You, Yanwu
中科院分区：
文献类型：
--
作者：
Gong, Qiming;Ma, Jingxue;You, Yanwu
背景足细胞损伤是原发性肾小球疾病的主要生物标志物，可导致大量蛋白尿和肾功能衰竭。趋化因子Fractalkine（FKN，CX 3CL 1）的产生增加是多种炎性疾病的标志。然而，FKN在足细胞损伤中的潜在机制仍然未知。方法建立FKN基因敲除（FKN-/-，FKN-KO）小鼠的LPS注射模型。在培养的足细胞中，我们使用质粒敲低FKN，并用PI 3 K/Akt抑制剂（LY 294002）处理足细胞。采用HE染色、Western Bolt、免疫共沉淀（Co-IP）、免疫荧光染色和流式细胞术分析FKN在足细胞损伤中的作用。结果与WT小鼠相比，LPS刺激导致肾脏损伤，Bcl-2家族凋亡蛋白表达增加，足细胞标志蛋白nephrin、podocin和WT 1丰度降低。与WT小鼠相比，LPS诱导的FKN-KO小鼠表现出降低的致死率和炎性细胞浸润、足细胞凋亡和PI 3 K/Akt信号通路抑制。在培养的足细胞中，FKN和PI 3 K/Akt信号通路之间的相互作用得到了很好的证实。FKN敲低通过调节Bcl-2家族减少足细胞凋亡;然而，这种保护作用被PI 3 K/Akt抑制剂（LY 294002）的共同给药逆转。结论FKN、PI 3 K/Akt信号通路和足细胞凋亡是一种新的机制。
Background Podocyte injury is a major biomarker of primary glomerular disease, which leads to massive proteinuria and kidney failure. The increased production of the chemokine, fractalkine (FKN, CX3CL1), is a hallmark of multiple inflammatory diseases. However, the underlying mechanism of FKN in podocyte injury remains unknown. Methods In this study, we performed an LPS infusion model in FKN knockout (FKN-/-, FKN-KO) mice. In cultured podocytes, we used plasmids to knockdown FKN and treated the podocytes with PI3K/Akt inhibitor (LY294002). Haematoxylin and eosin (HE) staining, Western Bolt, Co-immunoprecipitation (Co-IP), Immunofluorescence staining and flow cytometric analysis were employed to establish the role of FKN in podocyte injury. Results LPS stimulation resulted in kidney damage, increased the expression of the Bcl-2 family apoptosis protein, and decreased podocyte marker protein (nephrin, podocin and WT1) abundance compared with the WT mice. LPS-induced FKN-KO mice exhibited reduced lethality and inflammatory cell infiltration, podocyte apoptosis, and PI3K/Akt signal pathway inhibition compared to WT mice. In cultured podocytes, the interaction between FKN and the PI3K/Akt signalling pathway was well confirmed. FKN knockdown reduced podocyte apoptosis by regulating the Bcl-2 family; however, this protective effect was reversed by the co-administration of a PI3K/Akt inhibitor (LY294002). Conclusion Overall, these findings reveal a novel mechanistic property of FKN, PI3K/Akt signalling, and podocyte apoptosis.