RESUMO
Objective To investigate the potential of ginkgolide B(GB)in mitigating vascular endothelial injury by antagonizing endoplasmic reticulum stress(ERS)and elucidate its underlying molecular mechanism.Methods An injury model of human bone marrow-derived endothelial progenitor cells(EPCs)induced by tunica-mycin(TM)was established.Cell proliferation was assessed using MTS assay,while cell viability was determined through Calcein-AM/EthD-I double staining.Transwell assay was employed to evaluate cell migration ability.DCFH-DA staining was utilized to measure intracellular ROS levels,and NADPH activity was quantified via ELISA.JC-1 and DiOC6 staining were performed for qualitative and quantitative assessment of mitochondrial membrane potential respectively.Qrt-pcr analysis was conducted to determine mRNA expression levels,whereas western blot analysis enabled detection of protein expression levels in the cells.Results GB dose-dependently attenuated tunicamycin-induced ERS-mediated endothelial injury in hEPCs,as evidenced by decreased cell viability,impaired cell migration,and angiogenesis inhibition(P<0.01).Furthermore,GB treatment significantly reduced ROS production and NADPH levels within the cells(P<0.01),while also inhibiting ERS-mediated decline in mitochondrial membrane potential concentration-dependently(P<0.01).Additionally,GB inhibited the expression of ERS-related proteins such as GRP78,ATF4,CHOP etc.,regulated apoptosis-related protein Bcl-xl,Bax cleaved caspase-4 cytochrome c;thereby effectively counteracting endoplasmic reticulum stress-induced cellular damage.Conclusions GB exerts a protective effect on vascular endothelium by antagonizing endoplasmic reticulum stress;this mechanism may be attributed to its ability to reduce intracellular reactive oxygen species levels.It also suppresses the expression of ERS-related proteins(CHOP78 and ATF4),and modulates apoptosis-associated proteins(Bcl-xl,Bax,cleaved caspase-4,and cytochrome c).
RESUMO
Objective:To evaluate the protective immunity by vaccination of BALB/c mice with rLV-HA-GCN4,a recombinant lentivirus expressing the trimeric HA of swine H1N1 influenza virus. Methods:The female mice were randomly divided into rLV-HA-GCN4,rLV-HA,LV and PBS groups. Mice were primed with plasmid and boosted with lentivirus by the administration of intramuscular thigh injections at an interval of two weeks. At day 28 post-prime immunization,mice were inoculated intranasally with 100TCID50 of swine H1N1 influenza virus in a 50 μl volume. The immune levels were assessed by the T lymphocyte transformation test, flow cytometry,indirect ELISA and the indexes of spleen and lung. Results:The spleen lymphocyte transformation rate was 0. 3±0. 11 in the rLV-HA-GCN4 group at day 14 post-boost immunization, showing a statistical significance ( P<0. 01 ) compared to the PBS group. Meanwhile,rLV-HA-GCN4 could cause T lymphocyte response mainly based on the Th1-type CD4+ T cells. The IgG antibody titer reached to 1:8 000 at day 14 post-boost immunization and approximately 1:7 000 at day 14 post challenge. After challenge,the spleen and lung indexes of rLV-HA-GCN4 group were significantly lower than those of PBS group (P<0. 05). The body weight of rLV-HA-GCN4 group demonstrated a slight decrease before 3 days post challenge and then a gradual increase compared to the LV and PBS groups (P<0. 05). Conclusion:rLV-HA-GCN4 can effectively induce cellular and humoral immune response in BALB/c mice against swine H1N1 influenza virus.