Protective Effects Of Rutin On Cisplatin-induced Toxicity In Human Glomerular Mesangial Cell And Its Mechanisms

Backgroud:Cisplatin is one of the most effective agents widely used in the treatment of a variety of malignancies.Unfortunately,cisplatin also can cause a variety of side effects such as hepatotoxicity,renal toxicity and gonadal toxicity.Among the side effects,the incidence of renal toxicity was the most serious problem.Such side effects limit its clinical use.The mechanism of cisplatin-induced nephrotoxicity may be related to the accumulation of ROS and the activation of p53 and apoptosis factors.At present,the main clinical application of using hydrating agents to prevent cisplatininduced nephrotoxicity,but it may complicate the electrolyte disturbance.The rutin have antioxidant,anti-inflammatory,anti-allergic,anti-angiogenic and antiviral properties.In this study,human glomerular mesangial cells were selected as the study subjects.To investigate the protective effects of rutin on cisplatin-induced nephrotoxicity and its mechanisms.Purpose:By testing SOD,MDA,p53,pro-caspase-9,pro-caspase-3,cleaved-caspase-3,Bax and Bcl-2 expression in human glomerular mesangial cells to investigate the protective effect of rutin on cisplatin-induced nephrotoxicity.Methods:The cells were divided into five groups: control group;model group(DDP 10 ?g/ml)and DDP+RUT group.The DDP+RUT group was further divided into three different RUT dosage groups [DDP 10 ?g/ml + RUT 12.5 ?mol/L(Low dose)];DDP+RUT [DDP 10 ?g/ml + RUT 25 ?mol/L(Medium dose)];DDP+RUT [DDP 10 ?g/ml + RUT 50 ?mol/L(High dose)].The apoptotic rate,morphological change,active oxygen content,and content of SOD,MDA,p53 and pro-caspase-9,procaspase-3,cleaved-caspase-3,Bax and Bcl-2 in the cells were detected by MTT,fluorescence staining,reactive oxygenase,flow cytometry and Western Blotting.Results:1.MTT results showed that DDP could significantly decrease the viability of cells after 24 hours.rutin could improve cell viability at different concentrations.2.Cell morphology results showed that after 24 h treatment,the apoptotic cells in the model group were increased,the cells were dispersed and the cell disruption.The dosage of rutin was compared with the model group,the apoptotic cells were decreased and the cell morphology was significantly improved.3.Flow cytometry showed that the cells in the model group after 24 h treatment showed significant cycle arrest and the number of apoptotic cells increased.Compared with the model group,the cell cycle arrest and the number of apoptotic cells was decreased.4.The results of reactive oxygen species and SOD and MDA showed that the activity of reactive oxygen species increased,SOD activity decreased and MDA content increased after 24 h treatment.Compared with the model group,the content of reactive oxygen species decreased,SOD activity increased and MDA decreased.5.Western Blotting results showed that the expression of p53 and cleaved-caspase-3 were significantly increased in the model group,while the levels of pro-caspase-9,pro-caspase-3 and Bcl-2/Bax were significantly decreased.Compared with the model group,the expression of pro-caspase-9,pro-caspase-3 and Bcl-2 / Bax were significantly increased in the different dosage groups of rutin,and the expression of p53 and cleaved-caspase-3 were increased,especially in the middle dose.Conclusion:1.RUT can improve the cell survival rate and cell morphology.By increasing the activity of SOD in the cells,reducing the accumulation of ROS and MDA and improving the toxicity caused by DDP,it can proved that the RUTin can protect the HMCs anginst the cisplatin-induced nephrotoxicity.2.RUT can protects HMCs by downregulating the expression of related molecules in the p53 / caspase signaling pathway.