列当水提物对谷氨酸诱导的氧化衰老模型的影响及其机制
Effect and mechanism research of Orobanche cernua Loefling water extracts on glutamic acid-induced oxidative senescence model
侯婕 1刘鑫煜 1张鹏 1赵留威 1赵佳乐 1马环宇 1杨英 1高珍珍2
作者信息
- 1. 内蒙古农业大学兽医学院,内蒙古呼和浩特 010010
- 2. 内蒙古农业大学兽医学院,内蒙古呼和浩特 010010;江苏农林职业技术学院畜牧兽医学院,江苏句容 212499
- 折叠
摘要
探讨列当水提物(OLE)对谷氨酸(Glu)诱导的氧化衰老的影响及其机制.采用MTT法检测Glu、OLE对细胞增殖活力的影响;根据MTT试验结果确定最佳给药浓度与时间,设置空白组、Glu组(模型组)、OLE低、中、高质量浓度组(1.56、3.13、6.25 g/L).通过衰老相关β-半乳糖苷酶染色(SA-β-Gal)检测模型组与OLE组的衰老程度;检测各组超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GPX)活性及丙二醛(MDA)含量;通过流式细胞仪分析OLE作用12 h后细胞的周期分布;通过 Western blot检测Bcl-2、Bax、ERK1/2及磷酸化ERK1/2蛋白的相对表达水平.结果显示:与空白组相比,模型组SOD、GPX活性显著降低(P＜0.05),MDA含量显著上升,β-Gal染色阳性率显著上升,而经过OLE预处理再用Glu诱导组的MDA含量出现下降,SOD、GPX活性升高;与模型组相比,OLE预处理能抑制细胞凋亡(P＜0.05),作用于PC12细胞的G2期促进细胞增殖;与模型组相比,OLE中剂量组可显著提高ERK1/2、p-ERK1/2、Bcl-2的表达,降低Bax蛋白表达(P＜0.05).结果表明OLE可通过提高SOD、GPX活性,抑制MDA的产生,纠正Glu造成的氧化损伤,抑制细胞凋亡;通过提高ERK1/2、p-ERK1/2、Bcl-2的表达,降低Bax蛋白表达拮抗Glu带来的氧化损伤,对Glu引起的氧化衰老有预防作用.
Abstract
This study aims to explore the effects of aqueous Orobanche cernua Loefling extract(OLE)on glutamate(Glu)-induced oxidative senescence and its mechanism.The effects of Glu and OLE on cell proliferation viability were assessed using MTT assay;the optimal concentration and time of administration were determined based on the results of MTT assay,and experimental groups included the blank group,the Glu group(model group),and the groups with low,medium,and high quality concentrations of OLE(6.25,3.13,1.56 g/L).Staining for senescence-associated beta-galactosidase(SA-β-Gal)was used to detect the degree of senescence in the model and OLE groups;superoxide dismutase(SOD),glutathione peroxidase(GPX),and malondialdehyde(MDA)in each group were detected;flow cytometry was used to analyze the cell cycle distribution after 12 h of OLE treatment;and Western blot was used to detect the protein expression of Bcl-2,Bax,ERK1/2 and phosphorylated ERK1/2.The results showed that:compared with the blank group,the model group showed a significant decrease in SOD and GPX activities,a significant in-crease in MDA(P＜0.05),and a significant increase in SA-β-Gal staining positivity;compared with the blank group,OLE inhibited apoptosis(P＜0.05)and acted on the G2 phase of PC12 cells to promote cell proliferation;compared with the blank group,OLE medium-dose group significantly increased the expression of ERK1/2,p-ERK1/2,and Bcl-2,while decreasing the expression of Bax protein(P＜0.05).In conclusion,OLE may have a preventive effect on Glu-induced oxidative se-nescence by inhibiting the production of MDA,increasing the activities of SOD and GPX,and in-hibiting apoptosis.
关键词
列当/谷氨酸/氧化衰老/ERK1/2/Bax/Bcl-2Key words
Orobanche cernua Loef ling/glutamate/oxidative senescence/ERK1/2/Bax/Bcl-2引用本文复制引用
基金项目
国家自然科学基金地区科学基金(32060813)
出版年
2024
NETL
NSTL
万方数据