Sevoflurane Relieves Hepatic Ischemia–reperfusion Injury In Rats By Inhibiting The Expression Of Grp78

Objective This article aimed to study the role of sevoflurane pre-conditioning in hepaticischemia–reperfusion and its potential mechanism whether is related to the expression of Grp78.Methods A total of 24 male Sprague–Dawley rats with an average weight of 240 ±15 g.After a week of feeding,these rats were randomly divided into three groups: Sham group,I/R group,and SEV group.For rats of Sham group,after anesthesia,they were only underwent laparotomy and second laparotomy,while hepatic ischemia–reperfusion was not carried on them.Rat liver ischemia–reperfusion model was constructed in I/R group,and SEV group.For rats of SEV group,they were pre-treated with sevoflurane at first.After the operation,to collect three groups of rat vena cava blood samples,get the upper liquid centrifugal,liversamples were rapidly collected and prepared into 10%homogenate by using 0.9% saline.Serum TNF-?,IL-1?,IL-10,and IL-6 concentrations were detected by ELISA.Malondialdehyde(MDA),superoxide dismutase(SOD),and nitric oxide(NO)in liver homogenate were determined.Hematoxylin–Eosin staining,Tunel,and immunohistochemistry were performed.Ischemia–reperfusion hepatocyte model was established.Mouse embryonic hepatocytes BNLCL.2 were cultured and collected at the logarithmic growth stage.The cells were divided into Control group(Control group),model group(I/R group)and sevoflurane group(SEVgroup)according to different methods.Hepatic ischemia reperfusion model was established with saccharide-oxygen deprivation.SEVgroup was treated with sevoflurane in advance.Cells transfection was conducted.Normal BNLCL.2 cells were named as si Grp78 group and si NC group.For cells of I/R group,they were set as I/R+si NC group and I/R+ si Grp78 group.Meanwhile,cells of SEV group,they were renamed as I/R+si NC+SEV group and I/R+ si Grp78+SEV group.Apoptosis was observed by flow cytometry.Quantitative real-time PCR(q RT-PCR)to Grp78 m RNA and Western blotting analysis were used to calculate PERK,e IF2?,p-c-JNK/JNK.Results Compared with I/R group,liver damage degree,liver cell apoptosis,and glucoseregulatory protein 78(Grp78)expression was obviously reduced in rats of SEV group.TNF-?,IL-1?,and IL-6 concentrations were also significantly decreased(P<0.01).IL-10 was significantly higher.MDA and NO concentrations were dramatically lower(P<0.01)and SOD concentration was significantly higher(P<0.01).Apoptosis rate,Grp78,PERK,e IF2?,and p-c-JNK/JNK expression was also significantly decreased(P<0.01).si Grp78 m RNA and protein expression in BNLCL.2cells of si Grp78 group was significantly lower than that in si NC group(P<0.01).We also noted that,for cells of si NC group,I/R+si NC group and I/R+si NC+SEV group,the apoptosis rate of I/R+si NC group was higher than that of si NC group(P<0.01).Cells of I/R + si NC+SEV group were with much lower apoptosis rate when compared with I/R + si NC group(P<0.01).When compared with si Grp78 group,significantly increased apoptosis rate was found in I/R+si Grp78 group(P<0.01).However,no significant difference in apoptosis rate was found between I/R+si Grp78 group and I/R+si Grp78+SEV group.Western blotting analysis revealed significantly higher PERK,e IF2?,and p-c-JNK/JNK protein relative expression was found in I/R + si NC group when compared with si NC group(P<0.01).But these protein relative expression in I/R + si NC+SEV group was all markedly lower than those of I/R + si NC group(P<0.01).Meanwhile,it could also be noticed PERK,e IF2?,and p-c-JNK/JNK protein relative expression in I/R+si Grp78 group was much higher than those of si Grp78 group(P<0.01).However,when compared with I/R+si Grp78 group,changes of these protein relative expression in I/R+si Grp78+SEV group were not obvious.Sevoflurane significantly reduced apoptosis and expression of PERK,e IF2?,p-c-JNK/JNK by inhibiting the expression of Grp78(P<0.01).Conclusion Sevoflurane relieves hepatic ischemia–reperfusion injury by inhibiting the expression of Grp78.