RESUMO
Orientin is a flavonoid monomer. In recent years, its importance as a source of pharmacological active substance is growing rapidly due to its properties such as anti-myocardial ischemia, anti-apoptosis, anti-radiation, anti-tumor, and anti-aging. However, the neuroprotective effects of Orientin on stroke injury have not been comprehensively evaluated. The aim of the present study was thus to investigate the neuroprotective capacity and the potential mechanisms of Cyperus esculentus L. orientin (CLO) from Cyperus esculentus L. leaves against ischemia/reperfusion (I/R) injury using standard orientin as control. For in vitro studies, we treated HT22 cells with CoCl2 as an in vitro ischemic injury model. HT22 cells in the control group were treated with CoCl2. For in vivo studies, we used rat models of middle cerebral artery occlusion, and animals that received sham surgery were used as controls. We found that CLO protected CoCl2-induced HT22 cells against ischemia/reperfusion injury by lowering lipid peroxidation and reactive oxygen species formation as well as decreasing protein oxidation. However, CLO did not reduce the release of lactate dehydrogenase nor increase the activity of superoxide dismutase. Results showed that CLO could decrease neurological deficit score, attenuate brain water content, and reduce cerebral infarct volume, leading to neuroprotection during cerebral ischemia-reperfusion injury. Our studies indicate that CLO flavonoids can be taken as a natural antioxidant and bacteriostastic substance in food and pharmaceutical industry. The molecular mechanisms of CLO could be at least partially attributed to the antioxidant properties and subsequently inhibiting activation of casepase-3. All experimental procedures and protocols were approved on May 16, 2016 by the Experimental Animal Ethics Committee of Xinjiang Medical University of China (approval No. IACUC20160516-57).
RESUMO
AIM: To enhance the immunocontraceptive effect of Lagurus lagurus zona pellucida 3 DNA vaccine, and to achieve the prospect of application through the pVAX1-sig-LTB-lZP3-C3d3 different immunity pathway. METHODS: Two adjuvant molecules were constructed into the recombinant plasmid pVAX1-sig-LTB-lZP3-C3d3 as DNA vaccine which contains Escherichia coli heat-labile enterotoxin B subunit and the molecular adjuvant 3 copies of C3d. The results of RT-PCR and western blot showed that the DNA vaccine was expressed in mRNA and protein level. The female C57BL/6 mice were immunized by three ways: intramuscular injection, intranasal or oral route.Antibody levels and types were detected by ELISA. RESULTS: ELISA results showed that recombinant plasmid pVAX1-sig-LTB-lZP3-C3d3 immunization induced specific IgG, IgA levels were significantly different comparing with control (P<0.01). Antifertility experiment showed that the experimental group reduced the average fertility significantly different compared with the control group (P<0.01). CONCLUSION: Restriction analysis, RT-PCR and Western blot showed that the recombinant plasmid constructed correctly and can be the expression of mRNA and protein levels.It resulted that the recombinant plasmid pVAX1-sig-LTB-lZP3-C3d3 can induce the specific immune response efficiently and enhance the immunocontraceptive effects.
Assuntos
Proteínas do Ovo/genética , Proteínas do Ovo/imunologia , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/imunologia , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/imunologia , Vacinas Anticoncepcionais/genética , Vacinas Anticoncepcionais/imunologia , Vacinas de DNA/genética , Vacinas de DNA/imunologia , Animais , Anticorpos Neutralizantes/sangue , Arvicolinae , Anticoncepção Imunológica , Feminino , Vetores Genéticos/genética , Vetores Genéticos/metabolismo , Células HEK293 , Humanos , Imunoglobulina G/sangue , Infertilidade Feminina/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Glicoproteínas da Zona PelúcidaRESUMO
AIM: To investigated whether fai3 as molecular adjuvant can enhance antifertility effect of contraceptive DNA vaccine pcD-Lzp3 through intranasal route. METHODS: FAI was a fibrinogen-albumin-IgG receptor of group C streptococci in which exhibits adjuvant properties. The third gene fragment of FAI-fai3 (415~702 bp) has the function of mucosal adjuvant. In this study, Plasmid pcD-Lzp3and pcD-fai3were encapsulated separately or together with chitosan to generate chitosan-DNA complexes, chi-pcD-Lzp3, chi-pcD-fai3, and chi-(pcD-Lzp3+ pcD-fai3), as the DNA vaccines. ICR mice were immunized with these chitosan nanoparticles, respectively, on day 0, 14, 28 and 42. Indirect ELISA was employed to determine the anti-Lzp3 specific IgG, IgG1and IgG2a antibody in sera, and the anti-Lzp3 specific IgA in vaginal secretion and the feces. RESULTS: The results showed that higher levels of serum IgG and mucosal secreted IgA were induced significantly in the mice of intranasally co-administered with chi-(pcD-Lzp3+pcD-fai3). The antifertility results showed that the mice of co-immunized group had the lowest birth rate and mean litter size. CONCLUSION: The intranasal delivery of the molecular adjuvant fai3 with chi-pcD-Lzp3 could significantly enhance the antifertility of the Lzp3 DNA vaccine for contraception by enhancing both the systemic and mucosal immune responses.