Ulinastatin induces Nrf2/HO-1 axis and protects against oxidative stress in ovalbumin-induced murine asthma model / 中国药理学通报

ABSTRACT

Aim To explore the potential mechanism of ulinastatin’s antioxidant effect by examining the Nrf2 /HO-1 pathway.Methods OVA-induced asthma of mice was cured by intraperitoneal injection of ulinas-tatin (1 00 kU·kg -1 ·d -1 ).Control mice were given the same volume of PBS (pH 7.4).To investigate the effect of ulinastatin on airway hyperresponsiveness, levels of interleukin IL-4,IFN-γand OVA specific IgE in bronchoalveolar lavage fluid (BALF)were measured using enzyme-linked immunosorbent assays (ELISAs). The content of ROS from BALF of mice was tested in double hydrogen rhodamine (DHR)-1 23 method.The level of protein carbonyl and MDA from lung tissue of mice was detected with Protein carbonyl content assay kit and MDA kit.And antioxidative enzyme in mice BALF was tested by antioxidant enzyme kit.The levels of HO-1 in lung tissue from mice were detected by Western blot and Real-time PCR.Nuclear transfer and binding activity of Nrf2 were tested respectively by Western blot,IF and EMSA.Results Ulinastatin could alleviate the airway hyperresponsiveness,dis-tinctly reduce the content of IL-4,OVA specific IgE, ROS,protein carbonyl and MDA,but upraise the ex-pression of IFN-γand antioxidative enzyme such as SOD,GSH and TAOC. Moreover, the antioxidant effect of ulinastatin could be reversed by Znpp,which was the inhibitor of HO-1 .Ulinastatin could obviously induce the expression of HO-1 in protein level in a dose-and time-dependent manner.Ulinastatin could also induce the nuclear transfer of Nrf2 and increase the binding activity of Nrf2 as well as the expression of HO-1 in gene level;Conclusion Ulinastatin could induce the activation of Nrf2 /HO-1 pathway,which may contribute to the protective effects of ulinastatin a-gainst OVA-induced oxidative stress.