Adenoviral Transfer Of Hemopexin Gene Attenuates Oxidative Stress In Neurons Exposed To Blood Colt

A growing body of experimental evidence suggests that the heminwhich released from heme could accumulates in intracranial hematomasand a potent oxidant, the hemopexin(HPX) decreases hemin accumulationand catabolism by meural cells. In a prior study, we observed that HPXgene knockout increased the striatal injury and behavioral deficits of mousein intercerebral hemorrhage, To investigate the effect of HPX morespecifically, we extracted neuron-glia cells from cortex of newborn SD rats,transfer adenoviral hemopexin gene and then co-cultured primaryneuron-glia cell with50ul arterial blood blot using insert transwell, theblank group co-cultured with50ul DMEM/F12which content28ul serumwhile the control model group transfer adenoviral vector. By12h and24h,malondialdehyde (MDA) in supernatant was detected by Thibabituric Acidmethod, The HPX group MDA decreased compared with modelgroup(P<0.01), superoxide dismutase (SOD) was detected by TotalSuperoxide Dismutase Assay Kit with WST-1, and inducedglutathione(GSH) was detected by5,5’-dithio-bis(2-nitrobenzoic acid)(DNTB). SOD and GSH increased in HPX group compared withmodel group(P<0.05, P<0.01).The apoptosis of neural cells based onAnnexinV–fuoresce inisothiocyanate (FITC) and propidiumiodide(PI)stains turned no significant difference in HPX group compared with modelgroup(P>0.05) at12h, the percentage of apoptosis decreased at24h whileHPX group compared with model group(P<0.01). The expression of HO-1and caspase-3are detected by Western blot demonstrated that HO-1expression decreased in HPX group compared with control model group at24h (P<o.o1) while the caspase-3decreased in HPX group compared withmodel group both12h and24h(P<o.o1,P<0.05).All these result concludedthat hemopexin protects neuronal cell injury exposed to blood byanti-oxidation and decreasing HO-1and caspase-3expression.