López-Acosta Ocarol, Cristóbal-García Magdalena, Cardoso-Saldaña Guillermo, Carvajal-Aguilera Karla, El-Hafidi Mohammed
Depto. de Biomedicina Cardiovascular, Instituto Nacional de Cardiología Ignacio Chávez, Juan Badiano No 1, Colonia Sección XVI, Tlalpan 14080, Mexico.
Depto. de Endocrinología, Instituto Nacional de Cardiología Ignacio Chávez, Juan Badiano No 1, Colonia Sección XVI, Tlalpan 14080, Mexico.
Antioxidants (Basel). 2023 Mar 16;12(3):728. doi: 10.3390/antiox12030728.
The aim of this study was to evaluate the effect of leptin on reactive oxygen species' (ROS) generation of smooth muscle cells (SMCs) from a rat model of obesity and hyperleptinemia. Obesity and hyperleptinemia were induced in rats by a sucrose-based diet for 24 weeks. ROS generation was detected by using dichloro-dihydrofluorescein (DCF), a fluorescent ROS probe in primary SMCs culture. An increase in plasma leptin and oxidative stress markers was observed in sucrose-fed (SF) rats. At baseline SMCs from SF rats showed a more than twofold increase in fluorescence intensity (FI) compared to that obtained in control (C) cells. When the C cells were treated with 20 ng leptin, the FI increased by about 250%, whereas the leptin-induced FI in the SF cells increased only by 28%. In addition, sucrose feeding increased the levels of p22phox and gp91phox, subunits of Nox as an O source in SMCs. Treatment of cells with leptin significantly increased p22phox and gp91phox levels in C cells and did not affect SF cells. Regarding STAT3 phosphorylation and the content of PTP1B and SOCS3 as protein markers of leptin resistance, they were found to be significantly increased in SF cells. These results suggest that SF aortic SMCs are partially resistant to leptin-induced ROS generation.
本研究的目的是评估瘦素对肥胖和高瘦素血症大鼠模型中平滑肌细胞（SMC）活性氧（ROS）生成的影响。通过给予蔗糖基饮食24周诱导大鼠发生肥胖和高瘦素血症。在原代SMC培养物中，使用荧光ROS探针二氯二氢荧光素（DCF）检测ROS生成。在喂食蔗糖（SF）的大鼠中观察到血浆瘦素和氧化应激标志物增加。在基线时，与对照（C）细胞相比，SF大鼠的SMC荧光强度（FI）增加了两倍多。当用20 ng瘦素处理C细胞时，FI增加了约250%，而瘦素诱导的SF细胞中的FI仅增加了28%。此外，蔗糖喂养增加了SMC中作为氧源的Nox亚基p22phox和gp91phox的水平。用瘦素处理细胞可显著增加C细胞中p22phox和gp91phox的水平，而对SF细胞没有影响。关于STAT3磷酸化以及作为瘦素抵抗蛋白标志物的PTP1B和SOCS3的含量，发现它们在SF细胞中显著增加。这些结果表明，SF主动脉SMC对瘦素诱导的ROS生成具有部分抗性。
