RESUMO
A unique region of human parvovirus B19 virusVP1 (B19VVP1u) has been linked to a variety of cardiac disorders. However, the precise role of B19VVP1u in inducing cardiac injury remains unknown. The present study investigated the effects of B19VVP1u and different regions of B19VVP1u, including B19VVP1uA (residues 160), B19VVP1uB (residues 61129), B19VVP1uC (residues 130195) and B19VVP1uD (residues 196227), on inducing cardiac injury in naïve mice by zymography, immunoblotting, H&E staining and cytokine immunoassay. A significantly higher MMP9/MMP2 ratio and increased levels of inflammatory cytokines, including IL6 and IL1ß, were detected in the left ventricles of the mice injected with B19Vnonstructural protein 1 (B19VNS1) and B19VVP1u, accompanied by increased expression levels of phosphorylated (p)ERK and pP38. Significantly upregulated expression levels of atrial natriuretic peptide (ANP), hearttype fatty acidbinding protein (HFABP) and creatine kinase isoenzymeMB (CKMB), which are wellknown cardiac injury markers, as well as increased infiltration of lymphocytes, were detected in the left ventricles of the mice injected with B19VVP1, B19VNS1 and B19VVP1u. Moreover, a significantly higher MMP9/MMP2 ratio and increased levels of IL6 and IL1ß were observed in the left ventricles of the mice injected with B19VVP1u, B19VVP1uA, B19VVP1uB and B19VVP1uC, accompanied by upregulated pERK and pP38 expression. Notably, significantly lower levels of IL6 and IL1ß were observed in the left ventricles of the mice injected with B19VVP1uD. Furthermore, significantly increased ANP, HFABP and CKMB expression levels were detected in the left ventricles of the mice injected with B19VVP1u, B19VVP1uA and B19VVP1uB, along with enhanced infiltration of lymphocytes. Significantly higher serum IL1ß, IL6, TNFα and IFNγ levels were also detected in the mice injected with B19VVP1u, B19VVP1uA and B19VVP1uB. To the best of our knowledge, the findings of the present study were the first to demonstrate that the Nterminal region (residues 1129) of B19VVP1u induces an increase in the levels of cardiac injury markers, thus providing evidence for understanding the possible functional regions within B19VVP1u.
Assuntos
Proteínas do Capsídeo/imunologia , Traumatismos Cardíacos/imunologia , Infecções por Parvoviridae/complicações , Parvovirus B19 Humano/imunologia , Animais , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/isolamento & purificação , Citocinas/sangue , Citocinas/metabolismo , Modelos Animais de Doenças , Feminino , Traumatismos Cardíacos/sangue , Traumatismos Cardíacos/patologia , Traumatismos Cardíacos/virologia , Interações Hospedeiro-Patógeno/imunologia , Humanos , Camundongos , Infecções por Parvoviridae/sangue , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/virologia , Parvovirus B19 Humano/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/isolamento & purificação , Transdução de Sinais/imunologiaRESUMO
Astrocytes and astrocyte-related proteins play important roles in maintaining normal brain function, and also regulate pathological processes in brain diseases and injury. However, the role of astrocytes in the dopamine-depleted striatum remains unclear. A rat model of Parkinson's disease was therefore established by injecting 10 µL 6-hydroxydopamine (2.5 µg/µL) into the right medial forebrain bundle. Immunohistochemical staining was used to detect the immunoreactivity of glial fibrillary acidic protein (GFAP), calcium-binding protein B (S100B), and signal transducer and activator of transcription 3 (STAT3) in the striatum, and to investigate the co-expression of GFAP with S100B and STAT3. Western blot assay was used to measure the protein expression of GFAP, S100B, and STAT3 in the striatum. Results demonstrated that striatal GFAP-immunoreactive cells had an astrocytic appearance under normal conditions, but that dopamine depletion induced a reactive phenotype with obvious morphological changes. The normal striatum also contained S100B and STAT3 expression. S100B-immunoreactive cells were uniform in the striatum, with round bodies and sparse, thin processes. STAT3-immunoreactive cells presented round cell bodies with sparse processes, or were darkly stained with a large cell body. Dopamine deprivation induced by 6-hydroxydopamine significantly enhanced the immunohistochemical positive reaction of S100B and STAT3. Normal striatal astrocytes expressed both S100B and STAT3. Striatal dopamine deprivation increased the number of GFAP/S100B and GFAP/STAT3 double-labeled cells, and increased the protein levels of GFAP, S100B, and STAT3. The present results suggest that morphological changes in astrocytes and changes in expression levels of astrocyte-related proteins are involved in the pathological process of striatal dopamine depletion. The study was approved by Animal Care and Use Committee of Sun Yat-sen University, China (Zhongshan Medical Ethics 2014 No. 23) on September 22, 2014.