Carrasco-Pozo Catalina, Pastene Edgar, Vergara Carola, Zapata Moises, Sandoval Cristian, Gotteland Martin
Institute of Nutrition and Food Technology, University of Chile, Santiago, Chile.
Biochim Biophys Acta. 2012 Dec;1820(12):2052-61. doi: 10.1016/j.bbagen.2012.09.015. Epub 2012 Sep 26.
The effect of indomethacin (INDO) on Ca(2+) mobilization, cytotoxicity, apoptosis and caspase activation and the potential protective effect of quercetin (QUE), resveratrol (RES) and rutin (RUT) were determined in Caco-2 cells.
Caco-2 cells were incubated with INDO in the presence or absence of QUE, RES or RUT. The concentrations of Ca(2+) in the cytosol (Fluo-3 AM) and mitochondria (Rhod-2 AM) were determined as well as the cytotoxicity (MTT reduction and LDH leakage), apoptosis (TUNEL) and caspase-3 and 9 activities.
INDO promoted Ca(2+) efflux from the endoplasmic reticulum (ER), resulting in an early, but transient, increment of cytosolic Ca(2+) at 3.5min, followed by a subsequent increment of intra-mitochondrial Ca(2+) at 24min. INDO also induced cytotoxicity, apoptosis, and increased caspase activities and cytochrome c release. All these alterations were prevented by the inhibitors of the IP3R and RyR receptors, 2-Aminoethoxydiphenyl borate (2-APB) and dantrolene. QUE was the most efficient polyphenol in preventing Ca(2+) mobilization induced by INDO and all of its consequences including cytotoxicity and apoptosis.
In Caco-2 cells, INDO stimulates ER Ca(2+) mobilization, probably through the activation of IP3R and RyR receptors, and the subsequent entry of Ca(2+) into the mitochondria. Polyphenols protected the cells against the Ca(2+) mobilization induced by INDO and its consequences on cytotoxicity and apoptosis.
These results confirm the possibility of using polyphenols and particularly QUE for the protection of the gastroduodenal mucosa in subjects consuming NSAIDs.
在Caco-2细胞中测定了吲哚美辛（INDO）对Ca(2+)动员、细胞毒性、凋亡及半胱天冬酶激活的影响，以及槲皮素（QUE）、白藜芦醇（RES）和芦丁（RUT）的潜在保护作用。
在存在或不存在QUE、RES或RUT的情况下，将Caco-2细胞与INDO一起孵育。测定了胞质溶胶（Fluo-3 AM）和线粒体（Rhod-2 AM）中Ca(2+)的浓度，以及细胞毒性（MTT还原和LDH泄漏）、凋亡（TUNEL）和半胱天冬酶-3及-9活性。
INDO促进Ca(2+)从内质网（ER）流出，导致在3.5分钟时胞质Ca(2+)早期但短暂增加，随后在24分钟时线粒体内Ca(2+)增加。INDO还诱导细胞毒性、凋亡，并增加半胱天冬酶活性和细胞色素c释放。IP3R和RyR受体的抑制剂2-氨基乙氧基二苯硼酸盐（2-APB）和丹曲林可防止所有这些改变。QUE是预防INDO诱导的Ca(2+)动员及其所有后果（包括细胞毒性和凋亡）最有效的多酚。
在Caco-2细胞中，INDO可能通过激活IP3R和RyR受体刺激内质网Ca(2+)动员，随后Ca(2+)进入线粒体。多酚可保护细胞免受INDO诱导的Ca(2+)动员及其对细胞毒性和凋亡的影响。
这些结果证实了在服用非甾体抗炎药的受试者中使用多酚尤其是QUE保护胃十二指肠黏膜的可能性。
