| ObjectiveTo investigate whether curcumin combined with cisplatin can enhance the anti-tumor effect of cisplatin on A549 human lung adenocarcinoma cell,LLC-1mouse lung cancer cell and LLC-1 lung cancer mice,reduce the renal toxicity caused by cisplatin,and further explore its mechanism of action.MethodsIn vitro experiments: A549 and LLC-1 lung cancer cells were divided into Control group,DDP 13.3μmol group,DDP 13.3μmol + Cur 10μmol group,DDP13.3μmol + 20μmol/L group,DDP 13.3μmol + 40μmol/L group,DDP 13.3μmol+ 80μmol/L group,detected A549 and LLC-1 lung cancer cells viability by MTT;Applied mitochondrial membrane potential and PI-ANNEXIN V cell flow cytometry to detected the apoptotic rate of A549 and LLC-1 lung cancer cells;Applied cell migration assay to detect migration rate of A549 and LLC-1 lung cancer cells;Detection of inflammatory factors in A549 and LLC-1 lung cancer cell supernatants was by ELISA.In vitro experiments: A total of 48 male C57 mice were successfully inoculated with mouse-derived lung cancer cell LLC-1,divided into Control group,DDP 5mg/kg group,DDP combined with Low dose Cur(5mg/kg +50mg/kg),DDP combined with Middle dose Cur(5mg/kg + 100mg/kg),DDPwith High dose Cur(5mg/kg + 200mg/kg),DDP combined with P38 MAPK inhibitor(5mg/kg + SB 303580 1.4mg/kg).8 mice per group were observed for14 consecutive days.Measured tumor weight and calculated tumor inhibition rate.Blood cell count and renal function indicators were detected by automatic biochemical analyzer.HE staining was used to observe the tumor and kidney tissue.The changes of glutathione and malondialdehyde in kidney tissues were detected by ELISA;The expression of P38 and P-P38 in kidney tissues was detected by Western-blot method.ResultsIn vitro experiments: Compared with the Control group,DDP could inhibit the proliferation of A549 and LLC-1 lung cancer cells and reduce cell viability,mitochondrial membrane potential of A549 and LLC-1 lung cancer cells decreased,and the rate of apoptosis increased,the migration ability of A549、LLC-1 cancer cells decreased,at the same time,the contents of TNF-α and IL-6 in the supernatant of A549 and LLC-1 lung cancer cells decreased.Compared with the DDP group,the combined application of Cur and DDP could further enhance the inhibitory effect on A549 and LLC-1 lung cancer cells,reduce the mitochondrial membrane potential of A549 and LLC-1 lung cancer cells and promote apoptosis,inhibit the migration of A549 and LLC-1,reduce the levels of TNF-α and IL-6 in the supernatants of A549 and LLC-1lung cancer cells.In vivo experiments: Compared with the Control group,DDP could reduce the weight of lung cancer mice and inhibit tumor growth;HE staining of tumor and kidney tissues were found that tumor cell necrosis increased and kidney tissue structure was damaged;Various blood cell counts decreased,increased the level of serum creatinine and urea nitrogen;GSH content in kidney tissue decreased,and MDA content increased;P38 and P-P38 protein expression was up-regulated.Compared with the DDP group,the combined application ofCur and DDP could increase the weight of lung cancer mice and further increase the tumor suppression rate;HE staining of tumors and kidneys showed that the area of tumor necrosis increased,and the pathological damage of kidney tissues decreased;blood cell counts increased,serum creatinine and urea nitrogen content decreased;GSH content in kidney tissue increased,and MDA content decreased;And reduced the activity of P38 and P-P38 protein,inhibited the expression of P38,P-P38.ConclusionCur combined with DDP can enhance the growth inhibitory effect of DDP on A549,LLC-1 cells,promote the apoptosis of tumor cells,inhibit the migration ability and reduce the level of inflammatory factors in vitro.The combination of Cur and DDP can enhance growth inhibitory effect of DDP on C57 lung cancer mice in vivo,and reduce the nephrotoxicity and blood toxicity caused by DDP.The reduction of nephrotoxicity may be related to the down-regulated of P38 and P-P38 protein expression. |
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