[关键词]
[摘要]
探讨罗汉果粗提物(Momordica grosvenorii extract,MGE)对H2O2诱导MIN6细胞凋亡及氧化损伤的保护作用及机制。以不同浓度的H2O2诱导MIN6细胞建立氧化损伤模型,根据细胞活力选择最适的诱导浓度。采用MTT法检测细胞活力;流式细胞术检测细胞活性氧ROS水平和细胞凋亡率;Western blot方法对凋亡相关因子进行检测。结果表明,罗汉果粗提物(0~200 μg/mL)对MIN6细胞活性几乎无影响,因此选取200 μg/mL罗汉果粗提物进行后续实验。200 μg/mL罗汉果粗提物能显著逆转过氧化氢导致的细胞活力降低(p<0.05)。流式细胞术结果显示,与单用H2O2相比,罗汉果粗提物可以显著下调H2O2导致的ROS水平升高(p<0.05),且抑制H2O2诱导的MIN6细胞的凋亡。Western blot结果也表明罗汉果粗提物能部分逆转H2O2引起的PCNA表达降低,Bax表达增高(p<0.05)。综上所述,罗汉果粗提物可以抑制H2O2诱导MIN6细胞的ROS生成及细胞凋亡,该研究结果可为将罗汉果开发为防治糖尿病功能食品提供实验依据。
[Key word]
[Abstract]
The study is to investigate the protective effect and mechanism of crude Momordica grosvenorii extract (MGE) against hydrogen peroxide (H2O2)-induced apoptosis and oxidative damage in MIN6 cells. An oxidative damage cell model was established by the induction by H2O2 at different concentrations, and the optimal induction concentration was selected according to cell viability. The cell viability was determined by the MTT assay. Flow cytometry was used to detect the level of reactive oxygen species (ROS) and apoptosis rates of cells. The apoptosis-related factors were detected by Western blot. The results showed that the crude MGE (0~200 μg/mL) showed little effect on the viability of MIN6 cells, thus, 200 μg/mL of the crude Momordica grosvenorii extract was chosen for subsequent experiments. The crude MGE at 200 μg/mL significantly reversed the decrease (p<0.05) in H2O2-induced cell viability. The results of flow cytometry showed that compared with the treatment with H2O2 alone, the crude MGE decreased significantly (p<0.05) the H2O2-induced increase in ROS level, and inhibited H2O2-induced apoptosis of MIN6 cells. Moreover, the Western blot results revealed that the crude MGE could partially reversed H2O2-induced decrease in the expression of PCNA and increase in the expression of Bax (p<0.05). In summary, the crude MGE could inhibit H2O2-induced ROS generation and apoptosis in MIN6 cells. The results of this study can provide an experimental basis for the development of Momordica grosvenorii into functional foods for the prevention and treatment of diabetes.
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[基金项目]
广西省第四批创新驱动发展专项基金资助(桂科AA19254025);国家自然科学基金资助项目(81760663;31460229;82002822;81760443);中国博士后科学基金(2020M683630XB);广西科技计划项目基金资助(桂科ZD20302006);第四批八桂学者2017年专项经费([2017]143号);桂林市科技计划项目(20190206-1);“广西特聘专家”专项经费(2019B12);“广西医学高层次人才培养计划”(G202002005);广西壮族自治区卫生健康委员会广西神经鞘脂代谢相关疾病基础研究重点实验室(ZJC2020005)建设经费