Rutin Modulates MAPK Pathway Differently from Quercetin in Angiotensin II-Induced H9c2 Cardiomyocyte Hypertrophy

Abstract

Rutin is a flavonoid with antioxidant property. It has been shown to exert cardioprotection against cardiomyocyte hypertrophy. However, studies regarding its antihypertrophic property are still lacking, whether it demonstrates similar antihypertrophic effect to its metabolite, quercetin. Hence, this study aimed to investigate the effects of both flavonoids on oxidative stress and mitogen-activated protein kinase (MAPK) pathway in H9c2 cardiomyocytes that were exposed to angiotensin II (Ang II) to induce hypertrophy. Cardiomyocytes were exposed to Ang II (600 nM) with or without quercetin (331 μM) or rutin (50 μM) for 24 h. A group given vehicle served as the control. The concentration of the flavonoids was chosen based on the reported effective concentration to reduce cell hypertrophy or cardiac injury in H9c2 cells. Exposure to Ang II increased cell surface area, intracellular superoxide anion level, NADPH oxidase and inducible nitric oxide synthase activities, and reduced cellular superoxide dismutase activity and nitrite level, which were similarly reversed by both rutin and quercetin. Rutin had no significant effects on phosphorylated proteins of extracellular signal-related kinases (ERK1/2) and p38 but downregulated phosphorylated c-Jun N-terminal kinases (JNK1/2), which were induced by Ang II. Quercetin, on the other hand, had significantly downregulated the phosphorylated proteins of ERK1/2, p38, and JNK1/2. The quercetin inhibitory effect on JNK1/2 was stronger than the rutin. In conclusion, both flavonoids afford similar protective effects against Ang II-induced cardiomyocyte hypertrophy, but they differently modulate MAPK pathway.

Parkia speciosa Hassk. Empty Pod Extract Alleviates Angiotensin II-Induced Cardiomyocyte Hypertrophy in H9c2 Cells by Modulating the Ang II/ROS/NO Axis and MAPK Pathway

“…Cells were treated concurrently with Ang II (600 nM) ( Siti et al, 2021 ) and various concentrations of P. speciosa extract (3.125–100 μg/ml) for 24 h in eight-well chamber slides. Cell size was measured using immunofluorescence staining.…”

“…H9c2 cells were randomly assigned to seven groups: 1) control (vehicle), 2) 50 μg/ml P. speciosa extract, 3) Ang II (600 nM; Siti et al, 2021 ), 4) Ang II and 12.5 μg/ml extract, 5) Ang II and 25 μg/ml extract, 6) Ang II and 50 μg/ml extract, and 7) Ang II and 20 μM valsartan ( Al-Mazroua et al, 2013 ). Valsartan served as the positive control.…”

“…Cell size was measured following the method of Jeong et al (2015) with slight modifications described by Siti et al (2021) . Cells were stained with a primary antibody against α-actinin (1:200 dilution; ab9465, Abcam, Cambridge, MA, United States) followed by an Alexa Fluor 488-conjugated anti-mouse secondary antibody (1:200 dilution; A-11059, Invitrogen, Waltham, MA, United States) and visualized via fluorescence microscopy (Olympus Optical, Tokyo, Japan).…”

“…Protein expression was measured by Western blot as previously described ( Siti et al, 2021 ). Anti-phospho-ERK1/2 rabbit polyclonal (1:1,000) (#4377), anti-phospho-JNK1/2 rabbit monoclonal (1:1,000) (#4668), and anti-phospho-p38 mouse monoclonal (1:500) (sc-166182; Santa Cruz Biotechnology, Dallas, TX, United States) were the primary antibodies used in this study.…”

Interactions between the ERK1/2 signaling pathway and PCAF play a key role in PE‑induced cardiomyocyte hypertrophy

“…The development of cardiac hypertrophy is closely related to a variety of signaling pathways that interact with each other, for example, the Wnt signaling transduction pathway, the mitogen-activated protein kinase (MAPK) signaling transduction pathway and the microRNA signaling transduction pathway are all considered to be related to this process ( 3 , 6 , 7 , 9 ). Of these, the MAPK signaling pathway has attracted significant attention ( 43 , 44 ).…”