[Effects of Rg_1 on LPS-induced apoptosis and autophagy of lung epithelial cells].
[Effects of Rg_1 on LPS-induced apoptosis and autophagy of lung epithelial cells].
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DOI：
10.19540/j.cnki.cjcmm.20190111.002
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发表时间：
2019-04-01
期刊：
影响因子：
--
通讯作者：
Nie, Shi-Nan
中科院分区：
文献类型：
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作者：
Ji, Qi-Jian;Sun, Zhao-Rui;Nie, Shi-Nan
本研究旨在探讨人参皂苷Rg_1对内毒素（LPS）诱导的肺上皮细胞凋亡的保护作用及其机制。首先用LPS处理小鼠肺上皮细胞（MLE-12）。观察细胞自噬和凋亡的变化及其与LPS浓度和时间的关系。然后，在特定浓度和作用时间的LPS下，调节MLE-12细胞的自噬水平，观察细胞凋亡的变化。其次，在相同浓度和作用时间下，分别加入LPS诱导的自噬抑制剂3-MA和LPS诱导的Rg_1。将肺上皮细胞分组，观察LPS诱导的肺上皮细胞凋亡及抗凋亡药物Rg_1的作用机制。在动物实验中，将小鼠分组，通过凋亡蛋白、肺损伤评分和HE染色切片检测，验证抗LPS Rg_1是否对LPS诱导的肺损伤具有保护作用。结果表明，LPS作用12 h后，细胞凋亡和自噬均随LPS浓度的增加而增加。LPS浓度为25 g·L-1时，细胞凋亡逐渐增加，自噬随时间的推移先增加后减少。LPS组细胞凋亡高于对照组，LPS+3-MA组细胞凋亡进一步增加，而LPS+RAM（rapamycin，autophagy promoter）组细胞凋亡明显减少。LPS组自噬增加，LPS+3-MA组减少，LPS+RAM组增加。LPS组细胞凋亡率高于对照组，LPS+Rg_1组细胞凋亡率低于对照组。LPS+Rg_1+3-MA组凋亡细胞数再次增加。LPS组的自噬作用在给予LPS Rg_1后进一步增强，而在给予3-MA后减弱。在小鼠体内实验中，LPS组细胞凋亡率明显高于对照组，而LPS + Rg_1组细胞凋亡率明显低于对照组。肺损伤评分和HE染色也符合上述趋势。LPS可诱导肺上皮细胞凋亡，且呈时间和浓度依赖性。随着LPS浓度的增加，肺上皮细胞的自噬作用增强。在特定浓度的LPS下，自噬先增加，然后在12-16小时后减少。在一定时间内适当增加肺上皮细胞自噬可以减少LPS诱导的细胞凋亡，而抑制自噬可以增加细胞凋亡。人参皂苷Rg_1对自噬诱导的肺癌上皮细胞凋亡具有保护作用。
This paper aimed to study the protective effect of ginsenoside Rg_1 on endotoxin(LPS)-induced apoptosis of lung epithelial cells and its mechanism of action. Mouse lung epithelial cells(MLE-12) were first treated with LPS. The autophagy changes and apoptosis and the relationship with concentration and time of LPS were observed. Then,the level of autophagy in MLE-12 was regulated at a specific concentration and action time of LPS,and the changes of apoptosis were observed. Secondly,ginsenoside Rg_1 and autophagy inhibitor 3-MA were added respectively at the same concentration and action time of LPS. The lung epithelial cells were grouped to observe the effect of ginsenoside Rg_1 on LPS-induced apoptosis of lung epithelial cells and its mechanism. In the animal experiment,the mice were grouped and tested by apoptosis protein,lung injury score and HE staining section to verify whether ginsenoside Rg_1 has a protective effect on LPS-induced lung injury. The results showed that apoptosis and autophagy increased as the rise of concentration after treatment with LPS for 12 h. The apoptosis increased gradually,and the autophagy increased first and then decreased over time at the LPS concentration of 25 g·L-1. The apoptosis of LPS group was higher than that of control group,and LPS+3-MA group increased further,while apoptosis decreased significantly in LPS+RAM(rapamycin,autophagy promoter) group. The autophagy increased in LPS group,decreased in LPS+3-MA group and increased in LPS+RAM group. The apoptosis of LPS group was higher than that of control group,and the apoptosis of LPS+Rg_1 group decreased. The apoptosis of LPS+Rg_1+3-MA group increased again. The autophagy of LPS group further increased after administration of ginsenoside Rg_1,but decreased after administration of 3-MA. In the in vivo experiments in mice,the apoptosis of LPS group increased significantly compared with the control group,while LPS + ginsenoside Rg_1 group decreased. Lung injury score and HE staining also conformed to the above trend. LPS can induce the apoptosis of lung epithelial cells in a time-dependent and concentration-dependent manner. The autophagy of lung epithelial cells increases with the rise of LPS concentration. At the specific concentration of LPS,autophagy increases first and then decreases after 12-16 hours. Proper increase of autophagy in lung epithelial cells within a certain period of time can reduce the apoptosis induced by LPS,while inhibition of autophagy can increase apoptosis. Ginsenoside Rg_1 has a protective effect on lung cancer epithelial cell apoptosis induced by autophagy.