The Effect Of Nrf2 For The Damage Of TK6 Cell Induced By Pb(lead)

ObjectiveThe children’s immune system can be damaged by Pb exposure.The Nrf2 is a kind of protective protein molecular,which can protect cell from oxidative stress.We are studying the protective function of Nrf2 in TK6 cells damaged by Pb,hoping to understand the underlying mechanism of Nrf2 in preventing TK6 from toxic damage of Pb.The illumination of protective role of Nrf2 would be beneficial for therapy and prevention of Pb venenation.MethodsPart Ⅰ The underlying correlation ship between transcription factor Nrf2 and TK6 cytotoxicity induced by PbThe TK6 cells were respectively exposed to 0μM,116μM,233μM,466μM PbAc solution and positive group(H2O2).Then,the cell viability,oxidative stress and apoptosis of TK6 cells were respectively detected by relative biological kits.The RNA and protein of TK6 cells were extracted to get the information of gene expression of Nrf2,HO-1 and Nqo1 through qRT-PCR and Western-blot technology.Part Ⅱ The correlationship between transcription factor Nrf2 and the protection mechanism of Se for TK6 cytotoxicity induced by PbThe TK6 cells were respectively exposed to 0μM,116μM,233μM,466μM PbAc solution.Meanwhile,the TK6 cells were exposed to 116μM PbAc with 0.01μg Na2SeO3,233μM PbAc with 0.01μg Na2SeO3 and 466μM PbAc with 0.01μg Na2SeO3.Then,the viability,oxidative stress and apoptosis of TK6 cells were detected by relative biological kits.The RNA and protein of TK6 cells were extracted to get the information of gene expression of Nrf2,HO-1 and Nqo1 through qRT-PCR and Western-blot technology.ResultsPart Ⅰ The underlying correlation ship between transcription factor Nrf2 and TK6 cytotoxicity induced by Pb1.The results showed,compared with control group,the early apoptosis of high does,low dose and positive group were increased(P<0.05);The late apoptosis of high,medium,low dose group and positive group were increased(P<0.05),compared to control group.2.The ROS in high,medium,low dose group and positive group were significantly increased,compared to control group;The MDA in high,medium,low dose group and positive group were significantly increased,compared to control group(P<0.05);The SOD activity in high,medium,low dose group and positive group were significantly decreased,compared to control group(P<0.05);The ratio of GSH/GSSG in high,medium,low dose group and positive group were significantly decreased,compared to control group(P<0.05).3.Comparing with control group,the mRNA expression levels of Nrf2 in high,medium and low dose group were up-regulated(P<0.05);Except for low dose group,both of high and medium group respectively make the mRNA expression levels of HO-1 and Nqo1 up-regulated(P<0.05).4.According to the result,compared with control group,the cytoplasm protein levels of Nrf2 in high and medium group were up-regulated(P<0.05).But for the low does group,the the cytoplasm-protein levels of Nrf2 were down-regulated(P<0.05).Comparing with control group,the protein expression levels of nuclei-Nrf2 in high,medium and low dose group were up-regulated(P<0.05);Aside from low dose group,both of high and medium group induced the protein expression levels of HO-1 up-regulation(P<0.05);The protein expression levels of Nqo1 in high,medium and low dose group were up-regulated(P<0.05),when they were contrasted with control group.Part Ⅱ The correlationship between transcription factor Nrf2 and the protection mechanism of Se for TK6 cytotoxicity induced by Pb1.There was no any significantly difference in early apoptosis among all PbAc groups and PbAc with Na2SeO3 groups(P>0.05);The late apoptosis of TK6 cells were significantly different(P<0.05)between all PbAc groups and PbAc with Na2SeO3 groups.PbAc with Na2SeO3 groups can reduce the late apoptosis of TK6 cells,compared with PbAc groups(P<0.05).2.All PbAc with Na2SeO3 groups can decrease the ROS and MDA of TK6 cells,and promote the SOD activity of TK6 cells,compared to all PbAc groups(P<0.05).There was only significant difference in the ratio of GSH/GSSG in low PbAc groups and low PbAc with Na2SeO3 groups(P>0.05);3.The mRNA expression levels of Nrf2,HO-1 and Nqo1 were respectively up-regulated(P<0.05)in all PbAc with Na2SeO3 groups,compared to PbAc groups.4.The protein expression levels of cytoplasm-Nrf2 were no any significant difference in TK6 cells between all PbAc with Na2SeO3 groups and PbAc groups.Instead,the protein expression levels of nuclei-Nrf2 were significantly grown up in all PbAc with Na2SeO3 groups,compared to all PbAc groups;At the same time,the protein expression levels of HO-1and Nqo1 were up-regulated in all PbAc with Na2SeO3 groups,compared to all PbAc groups(P<0.05).Conclusions1.The apoptosis of TK6 cells can be induced for suffering from oxidative stress when it is exposed to Pb.2.The up-regulation of protein expression of Nrf2 can release effectively the damage of oxidative stress induced by Pb in TK6 cells.3.The protein expression of Nrf2 is extra up-regulated via Se to reduce the damage of oxidative stress,when it is in Pb exposure.4.The Nrf2 can protect TK6 cells from the damage induced Pb.