6-姜烯酚对H2O2诱导NCM460和HCT116氧化损伤的作用研究

Effects of 6-shogaol on H2O2-Induced Oxidative Damage in NCM460 and HCT116

摘要:

建立结直肠癌细胞氧化损伤模型，加入H2O2刺激，通过体外细胞实验研究6-姜烯酚对H2O2诱导人正常肠上皮细胞（NCM 460）和原位结肠癌细胞（HCT116）氧化损伤的不同作用及可能的分子机制。利用倒置显微镜观察不同浓度6-姜烯酚对H2O2诱导NCM460和HCT116后细胞形态的改变。CCK8法筛选6-姜烯酚的浓度区间，并测定细胞存活率。Annexin V-FITC/PI流式细胞术检测不同分组细胞凋亡。Western-blot检测分析相关凋亡蛋白（Caspase3、PARP1、MCC1、A2F、BCL2）的表达。结果显示，与对照组相比，6-姜烯酚可降低H2O2诱导的NCM460中Caspase3、PARP1、MCC1、A2F表达，促进BCL2的表达（p<0.05），具有抗氧化作用，并促进H2O2诱导的NCM460增殖（p<0.05），但是在HCT116中。6-姜烯酚却促进Caspase3、PARP1、MCC1、A2F表达，抑制BCL2表达（p<0.05），不同程度加强H2O2对HCT116的氧化损伤，抑制细胞增殖（p<0.05）。因此可以得出6-姜烯酚对H2O2诱导NCM460及HCT116具有明显不同的相反作用，这一发现可为进一步研究6-姜烯酚抗结直肠肿瘤具体相关机制或通路提供指导意义。

Abstract:

In this study, the model of oxidative damage in colorectal cancer cells was established. After the stimulation of hydrogen peroxide (H2O2), the different effects and possible molecular mechanisms of 6-shogoal on H2O2-induced oxidative damage in human intestinal epithelial cells (NCM460) and in situ colon cancer cells (HCT116) cells in vitro were investigated. Inverted microscope was used to observe the changes of cell morphology in HCT116 and NCM460 induced by H2O2 with different concentrations of 6-shogoal. CCK-8 (Cell Counting Kit-8) method was used to screen the concentration interval of 6-shogaol and the cell survival rate was determined. Annexin and V-FITC/PI cytometry were used to detect apoptosis of different groups. The expressions of related apoptotic proteins (Caspase-3, poly (ADP- ribose) polymerase-1 (PARP-1), MCC1, A2F, B-cell lymphoma-2 (Bcl-2)) were detected by Western-blot. As compared with the model group, 6-shogaol could reduce the expressions of caspase-3, PARP-1, MCC-1, A2F and promote the expression of Bcl-2 (p < 0.05) in NCM460 induced by H2O2, indicating that 6-shogaol had anti-oxidant and pro-proliferative effects (p<0.05). However, 6-shogoal could promote the expression of caspase3, PARP-1, MCC1 and A2F, and inhibit the expression of Bcl-2 (p<0.05) in HCT116 induced by H2O2, indicating that 6-shogaol could enhance the oxidative damage of HCT116 induced by H2O2 and inhibit cell proliferation (p<0.05). It could be concluded that 6-shogaol had different opposite effects in NCM460 and HCT116 induced by H2O2, which might provide guidance for further studies on specific mechanisms or pathways of 6-shogoal against colorectal cancer.