Effects of PHD and HSP90 on erythropoietin production in yak (Bos grunniens) renal interstitial fibroblast-like cells under hypoxia
Effects of PHD and HSP90 on erythropoietin production in yak (Bos grunniens) renal interstitial fibroblast-like cells under hypoxia
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PHD和HSP90对缺氧条件下牦牛肾间质成纤维细胞样细胞促红细胞生成素的影响
DOI：
10.1007/s10735-021-10054-6
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发表时间：
2022-01-27
影响因子：
3.2
通讯作者：
Bai, Xue-Feng
中科院分区：
文献类型：
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作者：
Cui, Yan;Li, Hui;Bai, Xue-Feng
促红细胞生成素（EPO）是红细胞生成的中心蛋白，在低氧适应过程中起重要作用，受低氧诱导因子（HIF）调节。然而，牦牛EPO产生细胞及其调控机制的研究尚未见报道。为了解牦牛促红细胞生成素（EPO）的产生及其调控，采集不同年龄牦牛肾脏，检测EPO、缺氧诱导因子1 α（HIF-1 alpha）和缺氧诱导因子2 α（HIF-2 alpha）的表达。分离培养肾小管上皮细胞（RTECs）和肾小管周间质成纤维细胞样细胞（RIFs），测定其产生EPO的能力。随后，用二甲基草酰甘氨酸（DMOG）和格尔德霉素（GA）处理细胞，其分别是脯氨酰-4-羟化酶结构域（PHD）和热休克蛋白90（HSP 90）的抑制剂，以及HIF-1 α和HIF-2 α的siRNA，以探索它们对EPO产生和调节的影响。结果表明，EPO、HIF-1 α和HIF-2 α在牦牛不同年龄组的表达量存在差异。高DMOG浓度引起RIF和RTEC中HIF-1 α和HIF-2 α水平的相应增加，然而，EPO水平仅在RIF中增加，并且在RTEC中在任何浓度下均未检测到;这表明EPO在RIF中产生。在用HIF-1 α和HIF-2 α的siRNA处理RIF后，我们发现EPO通过HIF-2 α被PHD调节。此外，增加GA浓度导致RIF中HSP 90、HIF-1 α、HIF-2 α和EPO的表达降低。总之，这些发现支持我们的主张，PHD调节EPO通过HIF-2 α在牦牛RIF，而HSP 90推动EPO的表达。
Erythropoietin (EPO), a central protein of erythropoiesis, plays an important role during hypoxia adaptation and is regulated by hypoxia-inducible factor (HIF). However, there is no report on EPO-producing cells and their regulatory mechanisms in yak (Bos grunniens). To understand EPO production and regulation of yak, kidneys from different age of yak were collected and expression of EPO, hypoxia-inducible factor 1 alpha (HIF-1 alpha), and hypoxia-inducible factor 2 alpha (HIF-2 alpha) were detected. Then renal tubule epithelial cells (RTECs) and peritubular interstitial fibroblast-like (RIFs) cells were isolated and cultured to determine their EPO production abilities. Subsequently, the cells were treated with dimethyloxalylglycine (DMOG) and Geldanamycin (GA), which are inhibitors of prolyl-4-hydroxylase domain (PHD) and heat shock protein 90 (HSP90) respectively, and siRNAs of HIF-1 alpha and HIF-2 alpha to explore their effect on EPO production and regulation. The results showed that expressions of EPO, HIF-1 alpha, and HIF-2 alpha were different in the different age groups of yak. High DMOG concentration caused a corresponding increase in the levels of HIF-1 alpha and HIF-2 alpha in RIFs and RTECs, however, EPO levels increased in RIFs only and was not detected at any concentration in RTECs; suggesting that EPO was produced in RIFs. Upon treating RIFs with siRNAs of HIF-1 alpha and HIF-2 alpha, we found that EPO was regulated by PHD through HIF-2 alpha. In addition, increasing GA concentration caused a decrease in expression of HSP90, HIF-1 alpha, HIF-2 alpha, and EPO in RIFs. In conclusion, these findings support our proposition that PHD regulates EPO via HIF-2 alpha in yak RIFs, while HSP90 impelled EPO expression.