HSP70into The Nucleus Combined With PARP-1Protection In Hepatic Ischemia-reperfusion Injury

Objective:Through the establishment of heat shock and hepatic ischemia-reperfusion model tostudy the role of HSP70into the nucleus binding with PARP-1in terms of hepaticischemia-reperfusion.Methods:Choose healthy male (Sprague Dawley SD) rats,weighing between180g-250g,wererandomly divided into five groups:HSP70inhibition group (H-/P+group); negativecontrol group (NC group); heat shock group(H+/P+group),to promote the expression ofHSP70by heat shock;positive control group (PCgroup),that ischemia-reperfusion group,Pringle was established by hepatic ischemia-reperfusion model; PARP-1inhibitinggroup(H+/P-group),3-AB inhibited by over-expression of PARP-1.After each group ofsuccessful modeling of liver tissue and blood samples were taken to analyze the survivalrate of rats,changes in liver function markers, HE staining of liver specimensmorphological changes, Western blot analysis of HSP70and PARP-1expression andimmuneco-precipitation analysis HSP70and the binding of PARP-1.The resultsusingprofessional image analysis software for statistical analysis using SPSS18.0software.Results:1. Automatic biochemical analyzer to detect serological samples,statistical softwareanalysis results are shown for each group were compared with each other significantdifference, P <0.01,statistically significant.ALT of PC group has highest value,there is asignificant difference compared with the other groups, P <0.01; H+/P+group and H-/P+group, H+/P-group there was a significant difference, P <0.01,ALT values lower; H-/P+group and the H+/P-compare the ALT values higher,there is a significant difference, P <0.01;indicate the nucleusof HSP70and PARP-1may play a liver protective effects,HSP70on liver function protective effect is stronger than PARP-1.2. By extracting the Nucleusline Westren Blot Detection Of HSP70, HSP70confirmed into The nucleus, and the clear expression of each treatment group of HSP70Statistical analysis showed that:H+/P+group and H+/P-have not significant difference,P>0.05; H-/P+group showed no significant difference with the PC group, P>0.05; H+/P+group, H+/P-compared with other groups,the expression of Hsp70increased,therewas a significant difference, P <0.01; NC group and the rest of the group weresignificantly different, P <0.01;only a small amount of Hsp70expression;indicate heatstress,ischemia-reperfusion injury could promote increased expression of HSP70,HSP70inhibition of expression of quercetinplayedthe.3.Extracting the nucleus of PARP-1line Westren blot detection, statistical analysis ofeach group of PARP-1expression: H+/P+hsa a significant difference compared withthe rest of the group, P <0.01,expression was significantly increased; H-/P+group andthe PC group,NC group and the H+/P-have significant difference increasing,P <0.01; H+/P-group, NC group PARP-1expression levels and more with the PC group,there was asignificant difference, P <0.01;there was a significant difference between PC andNCgroup,P <0.01show that the heat shock and ischemia-reperfusion injury haveprompted PARP-1protein expression increased,3-AB reachedinhibitingPARP-1overexpression.4.Co-immunoprecipitation tests: liver extract nuclear proteins in each group,respectively, with PARP-1, HSP70antibody immunoprecipitation, Western blot analysisagain, the results show: all appear in a band at70kDa and29kDa for HSP70and PARP-1proteins, HSP70was confirmed nucleus PARP-1and the presence of the complex.Indicates when the heat shock and ischemia-reperfusion injury and other stress, HSP70into the nucleus in combination with PARP-1.5.HE staining showed: NC group (control group) were normal lobular architecture,lobular central vein, hepatic sinusoidal structure is clearly visible, no obvious periportalinfiltration of neutrophils, no significant sinusoidal congestion; PC group(ischemia-reperfusion group) significant swelling of liver cells, sinusoidal narrow, congestion, lobular central venous congestion, liver sinusoidal cell cord and unclearboundaries, periportal see neutrophil infiltration; H-/P+group (HSP70inhibition group),H+/P-group (PARP-1inhibition group) and H+/P+pathological changes (heat shockgroup) liver cell damage between the NC group and the PC group.Conclusion:1.Heat and ischemia-reperfusion injury can promote the expression of HSP70andPARP-1ofcell.2.HSP70into the nucleus and binding wiht PARP-1protein,play a roll in protectingliver cells.