Department of Surgery, Chi Mei Medical Center, Tainan, Taiwan.
BMC Complement Altern Med. 2012 Aug 27;12:138. doi: 10.1186/1472-6882-12-138.
Doxorubicin (DOX) is an effective antineoplastic drug; however, clinical use of DOX is limited by its dose-dependent cardiotoxicity. It is well known that reactive oxygen species (ROS) play a vital role in the pathological process of DOX-induced cardiotoxicity. For this study, we evaluated the protective effects of guggulsterone (GS), a steroid obtained from myrrh, to determine its preliminary mechanisms in defending against DOX-induced cytotoxicity in H9C2 cells.
In this study, we used a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay, lactate dehydrogenase (LDH) release measurements, and Hoechst 33258 staining to evaluate the protective effect of GS against DOX-induced cytotoxicity in H9C2 cells. In addition, we observed the immunofluorescence of intracellular ROS and measured lipid peroxidation, caspase-3 activity, and apoptosis-related proteins by using Western blotting.
The MTT assay and LDH release showed that treatment using GS (1-30 μM) did not cause cytotoxicity. Furthermore, GS inhibited DOX (1 μM)-induced cytotoxicity in a concentration-dependent manner. Hoechst 33258 staining showed that GS significantly reduced DOX-induced apoptosis and cell death. Using GS at a dose of 10-30 μM significantly reduced intracellular ROS and the formation of MDA in the supernatant of DOX-treated H9C2 cells and suppressed caspase-3 activity to reference levels. In immunoblot analysis, pretreatment using GS significantly reversed DOX-induced decrease of PARP, caspase-3 and bcl-2, and increase of bax, cytochrome C release, cleaved-PARP and cleaved-caspase-3. In addition, the properties of DOX-induced cancer cell (DLD-1 cells) death did not interfere when combined GS and DOX.
These data provide considerable evidence that GS could serve as a novel cardioprotective agent against DOX-induced cardiotoxicity.
阿霉素（DOX）是一种有效的抗肿瘤药物；然而，其临床应用受到剂量依赖性心脏毒性的限制。众所周知，活性氧（ROS）在 DOX 诱导的心脏毒性的病理过程中起着至关重要的作用。在这项研究中，我们评估了乳香中提取的甾体化合物古尔胶素（GS）的保护作用，以确定其在防御 DOX 诱导的 H9C2 细胞毒性中的初步机制。
在这项研究中，我们使用 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基-2H-四唑溴盐（MTT）测定法、乳酸脱氢酶（LDH）释放测量和 Hoechst 33258 染色来评估 GS 对 DOX 诱导的 H9C2 细胞毒性的保护作用。此外，我们观察了细胞内 ROS 的免疫荧光，并通过 Western 印迹测量了脂质过氧化、caspase-3 活性和凋亡相关蛋白。
MTT 测定法和 LDH 释放表明，使用 GS（1-30 μM）治疗不会引起细胞毒性。此外，GS 以浓度依赖性方式抑制 DOX（1 μM）诱导的细胞毒性。Hoechst 33258 染色显示，GS 显著减少 DOX 诱导的凋亡和细胞死亡。使用 10-30 μM 的 GS 可显著减少 DOX 处理的 H9C2 细胞中细胞内 ROS 和上清液中 MDA 的形成，并将 caspase-3 活性抑制到参考水平。在免疫印迹分析中，GS 的预处理显著逆转了 DOX 诱导的 PARP、caspase-3 和 bcl-2 的减少，以及 bax、细胞色素 C 释放、裂解-PARP 和裂解-caspase-3 的增加。此外，当 GS 与 DOX 联合使用时，不会干扰 DOX 诱导的癌细胞（DLD-1 细胞）死亡的特性。
这些数据提供了充分的证据表明，GS 可以作为一种新型的心脏保护剂，用于预防 DOX 诱导的心脏毒性。
