摘要
研究麦冬皂苷D对麦冬皂苷D'诱导大鼠心肌细胞凋亡的干预作用及可能机制,为麦冬皂苷类成分的毒效差异深入研究提供参考。CCK-8法检测麦冬皂苷D与麦冬皂苷D'共处理对细胞存活率的影响; Western blot法及RT-PCR法检测麦冬皂苷D及麦冬皂苷D'对内质网应激相关基因PERK,Bip,ATF-4,p-eIF2α及CHOP表达的影响;流式细胞仪检测细胞凋亡率;通过内质网形态特异性示踪探针检测麦冬皂苷D'造成的内质网形态改变及麦冬皂苷D的逆转效果,探究麦冬皂苷D保护心肌细胞损伤的可能作用机制。细胞实验结果显示,6μmol·L-1的麦冬皂苷D'可显著诱导内质网应激相关蛋白的表达,导致细胞内质网形态发生变化,促进细胞凋亡。不同浓度的麦冬皂苷D可在一定程度上逆转麦冬皂苷D'引起的心肌细胞损伤。结果表明,麦冬皂苷D可降低麦冬皂苷D'触发的内质网应激,进而对心肌细胞产生保护作用。
This study is aimed to investigate the intervention effect and possible mechanism of ophiopogonin D( OPD) in protecting cardiomyocytes against ophiopogonin D’( OPD’)-induced injury,and provide reference for further research on toxicity difference of saponins from ophiopogonins. CCK-8 assay was used to evaluate the effect of OPD and OPD’ on cell viability. The effect of OPD on OPD’-induced cell apoptosis was measured by flow cytometry. Morphologies of endoplasmic reticulum were observed by endoplasmic reticulum fluorescent probe. PERK,ATF-4,Bip and CHOP mRNA levels were detected by Real-time quantitative polymerase chain reaction( PCR) analysis. ATF-4,phosphorylated PERK and e IF2α protein levels were detected by Western blot assay. Results showed that treatment with OPD’( 6 μmol·L^-1) significantly increased the rate of apoptosis;expressions of endoplasmic reticulum stress related genes were increased. The morphology of the endoplasmic reticulum was changed. In addition,different concentrations of OPD could partially reverse the myocardial cell injury caused by OPD’. The experimental results showed that OPD’-induced myocardial toxicity may be associated with the endoplasmic reticulum stress,and OPD may modulate the expression of CYP2 J3 to relieve the endoplasmic reticulum stress caused by OPD’.
作者
王佳
王宁宁
葛运炫
谭洪玲
马增春
王宇光
高月
WANG Jia;WANG Ning-ning;GE Yun-xuan;TAN Hong-ling;MA Zeng-chun;WANG Yu-guang;GAO Yue(Guangdong Pharmaceutical University,Guangzhou 510006,China;Institute of Radiation Medicine,Academy of Military Medical Sciences,Beijing 100850,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2019年第9期1876-1881,共6页
China Journal of Chinese Materia Medica
基金
国家自然科学基金项目(81873032)
