Junjie Wang1, Yan Jiang2, Senouthai Soulixay1, Dongdong Fu1, Yanwu You1. 1. Department of Nephrology, Affiliated Hospital of Youjiang Medical University for Nationalities, Baise 533000, China. 2. Science Lab Center, Youjiang Medical University for Nationalities, Baise 533000, China.
Abstract
OBJECTIVE: To investigate the role of Cyr61 in angiotensin Ⅱ (AngⅡ)-induced functional changes in HEK293 cells and explore the mechanism. METHODS: Cyr61 knockdown in cultured HEK293T cells was achieved by transfection of the cells with CRISPR/Cas9 KO plasmid. The changes in apoptosis and expression levels of Cyr61 and Bcl-2 in the cells with or without Cyr61 knockdown in response to treatment with 10-7 mol/L AngⅡ for 48 h were analyzed using flow cytometry, qRT-PCR and Western blotting. RESULTS: The cells with Cyr61 knockdown showed significantly decreased expression of Cyr61 protein as compared with the control cells (P < 0.05). AngⅡ treatment for 48 h significantly increased the expression of Cyr61 and lowers the expression of Bcl-2 at both the protein and mRNA levels in HEK293T cells. In HEK293T cells with Cyr61 knockdown, AngⅡ treatment resulted in significantly increased expression of Bcl-2 in HEK293T cells as compared with that of the control group (P < 0.05). AngⅡ treatment caused significantly increased apoptotic rate in HEK293T cells as compared with the cells with Cyr61 knockdown [(26.94 ± 3.73)% vs (3.87 ± 0.83)%, P < 0.05), and the apoptosis rate was significantly lowered to (15.76 ± 1.31)% in HEK293T cells with Cyr61 knockdown following AngⅡ treatment (P < 0.05). CONCLUSIONS: The up-regulation of Cyr61 expression is related with AngⅡ-induced injury in HEK293T cells, and down-regulating Cyr61 expression can effectively protect HEK293T cells against AngⅡ-induced injury.
OBJECTIVE: To investigate the role of Cyr61 in angiotensin Ⅱ (AngⅡ)-induced functional changes in HEK293 cells and explore the mechanism. METHODS:Cyr61 knockdown in cultured HEK293T cells was achieved by transfection of the cells with CRISPR/Cas9 KO plasmid. The changes in apoptosis and expression levels of Cyr61 and Bcl-2 in the cells with or without Cyr61 knockdown in response to treatment with 10-7 mol/L AngⅡ for 48 h were analyzed using flow cytometry, qRT-PCR and Western blotting. RESULTS: The cells with Cyr61 knockdown showed significantly decreased expression of Cyr61 protein as compared with the control cells (P &lt; 0.05). AngⅡ treatment for 48 h significantly increased the expression of Cyr61 and lowers the expression of Bcl-2 at both the protein and mRNA levels in HEK293T cells. In HEK293T cells with Cyr61 knockdown, AngⅡ treatment resulted in significantly increased expression of Bcl-2 in HEK293T cells as compared with that of the control group (P &lt; 0.05). AngⅡ treatment caused significantly increased apoptotic rate in HEK293T cells as compared with the cells with Cyr61 knockdown [(26.94 ± 3.73)% vs (3.87 ± 0.83)%, P &lt; 0.05), and the apoptosis rate was significantly lowered to (15.76 ± 1.31)% in HEK293T cells with Cyr61 knockdown following AngⅡ treatment (P &lt; 0.05). CONCLUSIONS: The up-regulation of Cyr61 expression is related with AngⅡ-induced injury in HEK293T cells, and down-regulating Cyr61 expression can effectively protect HEK293T cells against AngⅡ-induced injury.
Authors: Andres Hilfiker; Denise Hilfiker-Kleiner; Martin Fuchs; Karol Kaminski; Artur Lichtenberg; Hermann-Josef Rothkötter; Bernhard Schieffer; Helmut Drexler Journal: Circulation Date: 2002-07-09 Impact factor: 29.690
Authors: Roland Klingenberg; Soheila Aghlmandi; Christoph Liebetrau; Lorenz Räber; Baris Gencer; David Nanchen; David Carballo; Alexander Akhmedov; Fabrizio Montecucco; Stefan Zoller; Chad Brokopp; Dik Heg; Peter Jüni; Helena Marti Soler; Pedro-Manuel Marques-Vidal; Peter Vollenweider; Oliver Dörr; Nicolas Rodondi; François Mach; Stephan Windecker; Ulf Landmesser; Arnold von Eckardstein; Christian W Hamm; Christian M Matter; Thomas F Lüscher Journal: Eur Heart J Date: 2017-12-14 Impact factor: 29.983