Ilavenil Soundharrajan, Kim Da Hye, Jeong Young-Il, Arasu Mariadhas Valan, Vijayakumar Mayakrishnan, Prabhu Ponnuraj Nagendra, Srigopalram Srisesharam, Choi Ki Choon
Grassland and Forage division, National Institute of Animal Science, RDA, Republic of Korea-330-801.
The United Graduate School of Agricultural Sciences, Tottori University, Japan 680-8553.
Asian Pac J Trop Med. 2015 Apr;8(4):263-8. doi: 10.1016/S1995-7645(14)60328-X.
To elucidate the key parameters associated with hydrogen peroxide induced oxidative stress and investigates the mechanism of trigonelline (TG) for reducing the H2O2 induced toxicity in H9c2 cells.
Cytotoxicity and antioxidant activity of TG was assessed by EZ-CYTOX kit. RNA extraction and cDNA synthesized according to the kit manufacture protocol. Apoptosis was measured by the Flowcytometry, general PCR and qPCR.
It was found that the TG significantly rescued the morphology of the H9c2 cells. Treatment of cells with TG attenuated H2O2 induced cell deaths and improved the antioxidant activity. In addition, TG regulated the apoptotic gene caspase-3, caspase-9 and anti-apoptotic gene Bcl-2, Bcl-XL during H2O2 induced oxidative stress in H9c2 cells. These results were comparable with quercetin treatment. For evident, flow cytometer results also confirmed the TG significantly reduced the H2O2 induced necrosis and apoptosis in H9c2 cells. However, further increment of TG concentration against H2O2 could induce the necrosis and apoptosis along with H2O2.
It is suggested that less than 125 μ M of TG could protect the cells from H2O2 induced cell damage by down regulating the caspases and up regulating the Bcl-2 and Bcl-XL expression. Therefore, we suggest the trigonelline could be useful for treatment of oxidative stress mediated cardiovascular diseases in future.
阐明与过氧化氢诱导的氧化应激相关的关键参数,并研究胡芦巴碱(TG)减轻H2O2诱导的H9c2细胞毒性的机制。
使用EZ-CYTOX试剂盒评估TG的细胞毒性和抗氧化活性。按照试剂盒生产协议进行RNA提取和cDNA合成。通过流式细胞术、普通PCR和qPCR检测细胞凋亡。
发现TG显著挽救了H9c2细胞的形态。用TG处理细胞可减轻H2O2诱导的细胞死亡并提高抗氧化活性。此外,在H2O2诱导的H9c2细胞氧化应激过程中,TG调节凋亡基因caspase-3、caspase-9和抗凋亡基因Bcl-2、Bcl-XL。这些结果与槲皮素处理相当。明显地,流式细胞仪结果也证实TG显著降低了H2O2诱导的H9c2细胞坏死和凋亡。然而,TG浓度相对于H2O2的进一步增加会与H2O2一起诱导坏死和凋亡。
建议小于125μM的TG可通过下调半胱天冬酶和上调Bcl-2和Bcl-XL表达来保护细胞免受H2O2诱导的细胞损伤。因此,我们建议胡芦巴碱将来可能对治疗氧化应激介导的心血管疾病有用。