Protective effects of ulinastatin pretreatment regulating NLRP3 on neuron with hypoxic-ischemic damage

LI Xiaobing; YUE Zongyuan; BAI Jing; REN Lei; RAO Wei; PENG Cheng; FEI Zhou; ZHANG Lei

Abstract：Objective The potential therapeutic effect and the related mechanism of ulinastatin (UTI) and nucleotide-binding oligomerization domain receptor protein 3 (NLRP3) on cultured neuron associated with hypoxic-ischemic damage were investigated.Methods Neurons were in vitro cultivated.The cells were randomly divided into the sham group (Sham group),physiological saline control group (Control group),the damage group (OGD group) and the UTI pretreatment group (UTI group).Control group and UTI group were performed the pretreatment for 72 h,and then all the groups were subjected to oxygen-glucose deprivation (OGD) for 3 h and 6 h,lastly,all the neurons were cultivated by normal culture for another 2 h and then sampled.The expression of NLRP3 was detected by immunofluorescence staining;the expression of NLRP3 was detected by Western blotting.Neurons viability was detected by cell counting kit-8 (CCK-8).Results The expression of NLRP3 protein was different among Sham group,Control group,and UTI group (P ＜0.05);after OGD for 6 h,UTI could obviously reduce the expression of NLRP3 protein and the number of apoptotic neurons compared with Control group (P ＜ 0.05).But after OGD for 3 h,the expression of NLRP3 protein and the number of apoptotic neurons did no change significantly in UTI group compared with Control group (P ＞ 0.05).Conclusion The UTI pretreatmnent can effectively enhance the brain damage nerve function which maybe through regulating NLRP3 expression following the ischemic brain injury.