Effects Of Guiqi Yiyuan Gao On DNA Dsbs And Bystander Effect In Rats With Lung Injury Induced By Heavy-ion Rradiation

Objective: To observe the effects of Guiqi Yiyuan Gao on the signal factors,?H2AX and 53BP1,DNA double-strand damage and repair of the model rats of radiation-induced bystander effect of lung-kidney induced by heavy-ion irradiation on right lung;and to analyze the regulation mechanism of Guiqi Yiyuan Gao on DNA double-strand damage and repair of radiation-induced bystander effect of lung-kidney model rats;and to study its molecular biological mechanism to prevent RIBE on the level of DNA double-strand damage and repair.In the end,to provide the theoretical foundation and pharmacodynamic foundation for Guiqi Yiyuan Gao on preventing RIBE in the clinic.Methods: 42 Wistar rats of SPF level were randomly divided into normal control group,irradiation group,decoction group.rats of decoction groups were intervened by Guiqi Yiyuan Gao for 14 days,at the same time,other rats were intervened by normal saline.After the intervention,the right lung of rats of irradiation groups and decoction groups were irradiated by heavy ion and were killed after 6h,12 h,24h.Cell Cycle of Bone Marrow of rats were measured by Flow cytometry.The transcriptional level of H2 AX and 53BP1 were measured by Q-PCR,protein expression of ?H2AX and 53BP1 were measured by Western-blot.Results: 1.The changes of Cell Cycle of Bone Marrow:(1)Compared with normal control group,at G0/G1 phase,the cells in 12 hours after irradiation group were decreased(P<0.05);the cells in 12 hours after irradiation+decoction group and the cells in 24 hours after irradiation+decoction group were decreased(P<0.05).(2)Compared with normal control group,at S phase,the cells in 12 hours after irradiation group were increased(P<0.05);the cells in 6 hours after irradiation+decoction group,the cells in 12 hours after irradiation+decoction group and the cells in 24 hours after irradiation+decoction group was increased(P<0.05).(3)Compared with normal control group,at G2/M phase,cells in 12 hours after irradiation group and cells in 24 hours after irradiation group were increased(P<0.05);the cells in 24 hours after irradiation+decoction group were increased(P<0.05).2.The changes of transcriptional level of H2 AX of right lung,left lung and left kidney:(1)Compared with normal control group,transcriptional levels of H2 AX of right lung were increased in 6 hour,12 hour and 24 hour after irradiation and the difference were of statiscal significance(P<0.05).Compared with irradiation groups,transcriptional levels of H2 AX of right lung decreased in 6 hour,12 hour and 24 hour of irradiation+decoction groups(P<0.05).(2)Compared with normal control group,transcriptional levels of H2 AX of left lung were increased in 6 hour,12 hour and 24 hour after irradiation and the difference were of statiscal significance(P<0.05).Compared with irradiation groups,transcriptional levels of H2 AX of left lung decreased in 12 hour and 24 hour of irradiation+decoction groups(P<0.05).(3)Compared with normal control group,transcriptional levels of H2 AX of left kidney were increased in 6 hour,12 hour and 24 hour after irradiation and the difference were of statiscal significance(P<0.05).Compared with irradiation groups,transcriptional levels of H2 AX of left kidney decreased in 12 hour and 24 hour of irradiation+decoction groups(P<0.05).3.The changes of transcriptional level of 53BP1 of right lung,left lung and left kidney:(1)Compared with normal control group,transcriptional levels of 53BP1 of right lung were increased in 6 hour,12 hour and 24 hour after irradiation and the difference were of statiscal significance(P<0.05).Compared with irradiation groups,transcriptional levels of 53BP1 of right lung decreased in 6 hour,12 hour and 24 hour of irradiation+decoction groups(P<0.05).(2)Compared with normal control group,transcriptional levels of 53BP1 of left lung were increased in 6 hour,12 hour and 24 hour after irradiation and the difference were of statiscal significance(P<0.05).Compared with irradiation groups,transcriptional levels of 53BP1 of left lung decreased in 12 hour and 24 hour of irradiation+decoction groups(P<0.05).(3)Compared with normal control group,transcriptional levels of 53BP1 of left kidney were increased in 6 hour,12 hour and 24 hour after irradiation and the difference were of statiscal significance(P<0.05).Compared with irradiation groups,transcriptional levels of 53BP1 of left kidney decreased in 12 hour and 24 hour of irradiation+decoction groups(P<0.05).4.The changes of protein expression of ?H2AX of right lung,left lung and left kidney:(1)Compared with normal control group,protein expression levels of ?H2AX of right lung were increased in 6 hour,12 hour and 24 hour after irradiation and the difference were of statiscal significance(P<0.05).Compared with irradiation groups,protein expression levels of ?H2AX of right lung decreased in 6 hour,12 hour and 24 hour of irradiation+decoction groups(P<0.05).(2)Compared with normal control group,protein expression levels of ?H2AX of left lung were increased in 6 hour,12 hour and 24 hour after irradiation and the difference were of statiscal significance(P<0.05).Compared with irradiation groups,protein expression levels of ?H2AX of left lung decreased in 6 hour,12 hour and 24 hour of irradiation+decoction groups(P<0.05).(3)Compared with normal control group,protein expression levels of ?H2AX of left kidney were increased in 6 hour,12 hour and 24 hour after irradiation and the difference were of statiscal significance(P<0.05).Compared with irradiation groups,protein expression levels of ?H2AX of left kidney decreased in 6 hour,12 hour and 24 hour of irradiation+decoction groups(P<0.05).5.The changes of protein expression of 53BP1 of right lung,left lung and left kidney:(1)Compared with normal control group,protein expression levels of 53BP1 of right lung were increased in 6 hour,12 hour and 24 hour after irradiation and the difference were of statiscal significance(P<0.05).Compared with irradiation groups,protein expression levels of 53BP1 of right lung decreased in 6 hour,12 hour and 24 hour of irradiation+decoction groups(P<0.05).(2)Compared with normal control group,protein expression levels of 53BP1 of left lung were increased in 6 hour,12 hour and 24 hour after irradiation and the difference were of statiscal significance(P<0.05).Compared with irradiation groups,protein expression levels of 53BP1 of left lung decreased in 12 hour and 24 hour of irradiation+decoction groups(P<0.05).(3)Compared with normal control group,protein expression levels of 53BP1 of left kidney were increased in 6 hour,12 hour and 24 hour after irradiation and the difference were of statiscal significance(P<0.05).Compared with irradiation groups,protein expression levels of 53BP1 of left kidney decreased in 12 hour and 24 hour of irradiation+decoction groups(P<0.05).Conclusions: Guiqi Yiyuan Gao can effectively prevent double chain fracture of DNA and accelerate its repair at the same time,by directly adjust and control the expression of ?H2AX?53BP1 in directly irradiated structural right lung,structural left lung beside the directly irradiated and left kidney beside the directly irradiated.This is probably one of the mechanisms by which Guiqi Yiyuan Gao can avoid heavy-ion irradiation and its bystander effect.The paper provides new theoretical and pharmacodynamic foundation for the protection of the bystander effect damage of heavy-ion irradiation.