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Carbonic Anhydrase III Attenuates Hypoxia-Induced Apoptosis and Activates PI3K/Akt/mTOR Pathway in H9c2 Cardiomyocyte Cell Line
Cardiovascular Toxicology ( IF 3.2 ) Pub Date : 2021-08-13 , DOI: 10.1007/s12012-021-09683-w
Hua Li 1 , Yibin Liu 2 , Sha Tang 1 , Jie Hu 1 , Qiuling Wu 1 , Yang Wei 1 , Ming Niu 1
Affiliation  

Myocardial ischemia can cause insufficient oxygen and functional damage to myocardial cells. Carbonic anhydrase III (CAIII) has been found to be closely related to the abnormality of cardiomyocytes. To investigate the role of CAIII in the apoptosis of myocytes under hypoxic conditions and facilitate the strategy for treating hypoxia-induced damage, in vitro experiments in H9c2 were employed. The protein expression of CAIII in H9c2 cells after hypoxia or normoxia treatment was determined by western blotting and immunohistochemistry. MTT assay was employed for cells viability measurement and LDH release was monitored. The apoptotic cells were observed using immunofluorescence assay, flow cytometric analysis, and TUNEL assay. CAIII-overexpression or -knockdown cells were constructed to determine the role of CAIII in regulating apoptosis-related proteins caspase-3, Bax, Bcl-2, and anti-apoptosis pathway PI3K/Akt/mTOR. The mRNA levels of CAIII and genes related to CAIII synthesis including REN, IGHM, APOBEC 3F, and SKOR2 were significantly upregulated in hypoxia fetal sheep. The expression of CAIII protein and content of apoptotic H9c2 cells were increased at 1, 3, 6, and 12 h after hypoxia treatment. Overexpression of CAIII significantly upregulated Bcl2 level and downregulated Bax and caspase-3 cleavage levels, while its knockdown led to the contrary results. Overexpressed CAIII promoted the HIF-1α level and activated the PI3K/Akt/mTOR pathway, thereby exerting an inhibitory effect on hypoxia-induced apoptosis. In conclusion, our findings revealed that CAIII could protect cell from hypoxia-apoptosis of H9c2 cells, in which, activated PI3K/Akt/mTOR signaling pathway may be involved.



中文翻译:

碳酸酐酶 III 减弱缺氧诱导的细胞凋亡并激活 H9c2 心肌细胞系中的 PI3K/Akt/mTOR 通路

心肌缺血会导致心肌细胞供氧不足和功能损伤。已发现碳酸酐酶III(CAIII)与心肌细胞的异常密切相关。为了研究CAIII在缺氧条件下肌细胞凋亡中的作用并促进治疗缺氧诱导的损伤的策略,采用了H9c2的体外实验。通过蛋白质印迹和免疫组织化学测定缺氧或常氧处理后H9c2细胞中CAIII的蛋白表达。MTT测定用于细胞活力测量并监测LDH释放。使用免疫荧光法、流式细胞仪分析和TUNEL法观察凋亡细胞。构建 CAIII 过表达或敲低细胞以确定 CAIII 在调节凋亡相关蛋白 caspase-3、Bax、Bcl-2 和抗凋亡途径 PI3K/Akt/mTOR 中的作用。CAIII 的 mRNA 水平和与 CAIII 合成相关的基因包括 REN、IGHM、APOBEC 3F 和 SKOR2 在缺氧胎羊中显着上调。缺氧处理后1、3、6、12 h CAIII蛋白表达和凋亡H9c2细胞含量增加。CAIII 的过表达显着上调 Bcl2 水平并下调 Bax 和 caspase-3 切割水平,而其敲低导致相反的结果。过表达的 CAIII 促进 HIF-1α 水平并激活 PI3K/Akt/mTOR 通路,从而对缺氧诱导的细胞凋亡发挥抑制作用。综上所述,

更新日期:2021-08-19
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