| Objective:To investigate whether naringenin(Nar)regulates the proliferation and apoptosis of lung cancer A549 cells by acting on ROS/P38 MAPK signaling pathway.Methods: Culture A549 cells as experimental subjects.1.The control group and 30、60、90 and 120μmol/L Nar groups were set up,and different concentrations of Nar were added to A549 cells for 48 h to observe the changes in cell growth in each group.The cell vitality of A549 cells in the above groups were detected by CCK-8 method,and the direct effect of Nar on normal proliferation of A549 cells was observed,and follow-up experiments were prepared.2.The groups were the same as above.Cell apoptosis and ROS content were detected by flow cytometry.3.The groups were the same as above.The activity of SOD in each group was detected by WST-8 method and the content of MDA in each group was detected by TBA method.4.Set up the control group and different concentration Nar groups,and the inhibitor group,a total of six groups,respectively for 30、60、90、120μmol/L Nar,and 120μmol/L Nar+ P38 MAPK inhibitor 10μmol/L processing after 48 hours,using Western blot method to detect Nrf2 and downstream NQO1、HO-1 protein expression level,and detection of P38 MAPK protein phosphorylation and caspase-3activation levels,above analysis of the change of protein expression,To investigate the effect of Nar on the proliferation and apoptosis of A549 cells.Results:1.Compared with the control group,lung cancer A549 cells were treated with different concentration gradient of Nar(30、60、90、120μmol/L),the proliferation of A549 cells was inhibited in different concentrations of Nar groups,and the proliferation inhibition was more obvious with the increase of Nar concentration(P<0.05).Analysis of variance showed that the survival rate of the five groups was statistically significant(F =70.782,P<0.001).2.Compared with the control group,A549 cells were treated with different concentration gradients of Nar,with the increase of Nar concentration,the apoptosis of A549 cells was gradually increased(P<0.001),and the difference between the five groups was statistically significant by ANOVA(P<0.001).Compared with the control group,the expression of MFI in naringin group with different concentration gradient was increased(P<0.001),and with the increase of Nar concentration,The MFI value increased significantly.The difference among the five groups was statistically significant(F=178.220,P<0.001).3.Compared with the control group,SOD activity in Nar groups with different concentration gradient decreased(P<0.05),and SOD activity showed an obvious trend of decrease with the increase of Nar concentration.ANOVA showed a statistically significant difference among the five groups(F=76.847,P=0.000).Compared with blank control group,MDA content in Nar groups with different concentrations increased(P<0.05),and MDA content showed a significant increasing trend with the increase of Nar concentration.ANOVA showed a statistically significant difference among the 5 groups(F=50.589,P=0.000).4.Compared with the control group,the expression levels of Nrf2、NQO1 and HO-1 in 30、60、90 and 120μmol/L Nar groups were significantly decreased with the increase of Nar concentration,and the protein expression was significantly decreased in the 5 groups.5.Compared with the control group,the phosphorylation level of P38 MAPK protein and the activation level of caspase-3 protein in 30、60、90 and 120μmol/L Nar groups were significantly increased with the increase of Nar concentration,and the differences among the five groups were statistically significant by ANOVA(F=108.073,P<0.001).Compared with 90μmol/L and 120μmol/L Nar groups,P38 MAPK protein phosphorylation and caspase-3 activation were decreased in the inhibitor group(P<0.05),while there was no statistical significance in the inhibitor group compared with 30μmol/L and 60μmol/L Nar groups(P>0.05).Conclusion:1.The proliferation activity of A549 cells was inhibited by Nar,and the inhibitory effect of Nar on A549 cells was enhanced with the increase of Nar dose,and the effect was strongest in the 120μmol/L Nar group.2.Nar can significantly promote apoptosis of A549 cells,which can cause oxidative stress damage in a concentration-dependent manner.With the increase of Nar concentration,the apoptosis of A549 cells also increases gradually.3.Nar can inhibit the proliferation and promote apoptosis of A549 cells,and the reaction mechanism may be related to the activation of ROS/p38-MAPK signaling pathway. |
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