Cardioprotective effects of rhamnetin in H9c2 cardiomyoblast cells under H₂O₂-induced apoptosis

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• 作者：Eun-Seok Park^a ; Jun Chul Kang^a ; Yong Chang Jang^a ; Jong Seok Park^b ; Shin Yi Jang^c ; Dae-Eun Kim^d ; Bokyung Kim^e ; Hwa-Sup Shin^a ; ^{hsshin@kku.ac.kr" class="auth_mail}
• 关键词：CAT ; catalase ; DMEM ; Dulbecco壮s modified Eagle壮s media ; DMSO ; dimethylsulfoxide ; ERK ; extracellular signal regulated kinase ; FBS ; fetal bovine serum ; JNK ; c-Jun N-terminal protein kinase ; MAPKs ; mitogen-activated proteins kinases ; XTT ; 2 ; 3-bis [2-methyloxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide ; PBS ; phosphate-buffered saline ; ROS ; reactive oxygen species ; SDS-PAGE ; sodium dodecyl sulfate-polyacrylamide gel electrophoresis ; SOD ; superoxide dismutase ; TUNEL ; terminal deoxynucleotid
• 刊名：Journal of Ethnopharmacology
• 出版年：14 May 2014
• 年：2014
• 卷：153
• 期：3
• 页码：552-560
• 全文大小：1956 K

文摘

Many studies have emphasized that flavonoids, found in various fruits, vegetables, and seeds, as well as tea and red wine, have potential health-promoting and disease-preventing effects. Rhamnetin is a flavonoid that exhibits antioxidant capabilities. However, little is known about its effect on cardiac myocytes under oxidative stress and the underlying mechanisms.

Materials and methods

H9c2 cardiomyoblast cells were subjected to H₂O₂, to study the protective effect of rhamnetin on cell viability, apoptosis, and ROS production. Signaling proteins related to apoptosis, survival, and redox were analyzed by Western blot. Furthermore, the mRNA expressions of SIRTs were tested by real time-polymerase chain reaction (PCR).

Results

We investigated the protective effects of rhamnetin against H₂O₂-induced apoptosis in H9c2 cardiomyoblasts. Rhamnetin protected cells against H₂O₂-induced cell death without any cytotoxicity, as determined by the XTT assay, LDH assay, TUNEL assay, Hoechst 33342 assay, and Western blot analysis of apoptosis-related proteins. Rhamnetin also enhanced the expression of catalase and Mn-SOD, thereby inhibiting production of intracellular ROS. Furthermore, rhamnetin recovered the H₂O₂-induced decrease in phosphorylation of Akt/GSK-3尾 and MAPKs (ERK1/2, p38 MAPK, and JNK) and pretreatment with their inhibitors, attenuating the rhamnetin-induced cytoprotective effect. Further studies with real time-PCR and a sirtuin inhibitor showed that cardioprotection by rhamnetin occurred through induction of SIRT3 and SIRT4.

Conclusions

Taken together, these results suggest that rhamnetin may have novel therapeutic potential to protect the heart from ischemia-related injury.