Effect Of Nano-cadmium Sulfide On Vitro Toxicity Expression In Human Renal Tubular Epithelial Cells

ObjectiveTo study the cytotoxicity of Nano Cadmium sulfide(nCdS)on human renal tubular epithelial cells(HKC),and to study the oxidative damage,inflammatory response and related mechanisms.Method1)Nano Cadmium sulfide(nCds)characterization: using transmission electron microscopy(TEM)and X-ray diffraction(XRD)nanoparticles used in experiments were carried out to characterize,reach the size of 6 ~ 8nm can be used for this study.2)Culture of HKC:Cells were cultured and identified in vitro.3)Nano Cadmium sulfide(nCds)on HKC growth: CdS nanoparticles were treated with different doses of 24h(nCds),and negative control group,using the method of MTT(different dose group)detection of Nano Cadmium sulfide nanoparticles(nCds)on cell influence.4)nCds on HKC were studied: CdS nanoparticles with different doses of 24h(nCds),and negative control group,using AO/EB staining,DAPI staining changes in cell morphology were observed under fluorescence microscope,transmission electron microscope to observe the changes of nucleus.5)Effects of nCds on oxidative stress in cultured HKC: detection of ROS and GSH in cells treated with nCds.6)nCds on HKC to explore the internal mechanism of oxidative stress: immunohis to chemical observation of nano Cadmium sulfide(nCds)on cell MDA,SOD,GSH-Px,GSH the influence of Uchiya Ka cells the relative expression of cells,quantitative real-time PCR to detect GSH-Px,SOD,MDA,GSH.7)nCds on HKC to investigate the inflammatory reaction in the mechanism: by Western blotting(Western blotting method)to detect the Nano Cadmium sulfide(nCds)effect of NF-kappa B protein expression level in the treated cells.ResultWith the development of nCds dose increased,HKC survival rate was gradually decreased,ROS increased,MDA decreased,GSH-Px content,GSH content,the intra and external cellular Cd2+ concentration increased.The survival rate of HKC 10,20,40,80g/mL dose group decreased compared with the control group,the difference was statistically significant(P<0.05).10,20,30,40 g/m L dose group of ROS and MDA content as well as the 40 g/m L dose group of GSH and control group difference was statistically significant(P <0.05).With the development of nCds dose increased,compared with SOD,ROS,MDA,GSH-Px,GSH and control group,reduce the content of 40 g/m L dose group of SOD,GSH-Px,GSH,increase the content of ROS,MDA,(P <0.05).Cadmium sulfide(nCdS)on the expression of NF-kappa B protein level in HKC cells,in addition to the 10 g/m L dose group had no significant difference,other each treatment group were significantly higher than the control group(P<0.05),and with the increase of treatment dose,the expression of NF-kappa B protein levels increased gradually.Conclusion1 Nano Cadmium sulfide(nCds)can reduce the survival rate of human renal tubular epithelial cells(HKC),ROS and NF-kappa B protein increased,SOD,MDA,GSH-Px,GSH.2 This study tested the toxicity of nCds is produced by the effect of reactive oxygen species and inhibit the antioxidant and oxidative stress leads to mitochondrial dysfunction,which caused different changes in gene expression.3 In this study,we verified that the toxicity of n Cds by the NF-kappa B and other related proteins.4 The mechanism of oxidative stress,inflammatory reaction and apoptosis related pathway in the mechanism of nCds was investigated.