Angiotensin II type 1 receptor-associated protein plays a role in regulating the local renin-angiotensin system in HSC-T6 cells
Angiotensin II type 1 receptor-associated protein plays a role in regulating the local renin-angiotensin system in HSC-T6 cells
复制标题
DOI：
10.3892/mmr.2015.3849
复制
发表时间：
2015-09-01
影响因子：
3.4
通讯作者：
Hu, Yunwei
中科院分区：
文献类型：
--
作者：
Huang, Haiyang;Zhou, Jie;Hu, Yunwei
本研究的目的是检测血管紧张素II 1型受体相关蛋白(ATRAP)在大鼠肝星状细胞系HSC-T6中的表达，并确定其与局部肾素-血管紧张素系统(RAS)的相互作用。为达到这一目的，观察血管紧张素II(AngII)和血管紧张素-(1-7)[Ang-(1-7)]刺激HSC-T6细胞后2、6、12、18、24和36h对ATRAP、血管紧张素II I型受体(AT1R)、Mas受体和血管紧张素转换酶2(ACE2)表达的影响。用RT-qPCR和Western blotting分别在基因和蛋白水平对表达的变化进行量化。单次给予血管紧张素转换酶抑制剂(1mU/L)后12h，血管紧张素转换酶基因表达显著增加，而血管紧张素转换酶2基因表达水平在6h显著升高，12h后恢复至基础水平，然后显著降低。与AngII相同浓度的Ang-(1-7)可诱导ATRAP基因表达，在6h时点有统计学意义，12h达高峰，并在整个实验过程中持续升高。Ang-(1-7)可显著抑制血管紧张素转换酶2(ACE2)mRNA的表达，24 h达最低水平，而AT1R和Mas受体的表达不受Ang-(1-7)和Ang-(1-7)刺激。Western blotting结果与基因表达数据基本一致。综上所述，ATRAP在HSC-T6细胞中内源性表达，因此在调节HSC-T6细胞中的局部RAS方面可能起关键作用。
The aim of the current study was to examine the expression of the angiotensin II type 1 receptor-associated protein (ATRAP) in the rat hepatic stellate cell line HSC-T6 and to determine its interactions with the local renin-angio-tensin system (RAS). To achieve this goal, the effect of stimulating HSC-T6 cells with angiotensin II (AngII) and angiotensin-(1-7) [Ang-(1-7)], on the expression of ATRAP, the angiotensin II type I receptor (AT1R), the Mas receptor and the angiotensin converting enzyme 2 (ACE2) 2, 6, 12, 18, 24 and 36 h after stimulation was investigated. Changes in expression were quantified at the gene and protein level using RT-qPCR and western blotting, respectively. A single dose of AngII (1 mu mol/l) significantly increased the gene expression of ATRAP at 12 h, whereas ACE2 gene expression levels were significantly increased at 6 h and then returned to baseline at 12 h, prior to becoming significantly lower. A single dose of Ang-(1-7) at the same concentration as AngII induced ATRAP gene expression, which became statistically significant at the 6 h time-point, reached a peak at 12 h and remained elevated throughout the experimental time-course. In addition, ACE2 mRNA expression was significantly suppressed by Ang-(1-7) at 6 h, reaching its lowest expression level at 24 h. The expression of AT1R and the Mas receptor were unaffected by stimulation with AngII and Ang-(1-7). The western blotting results were generally consistent with the mRNA expression data. In conclusion, it was identified that ATRAP is endogenously expressed in HSC-T6 cells and therefore, may be critical in regulating the local RAS in these cells.