RESUMO
Previous research has focused on revealing the functions of each individual gene and/or pathway in idiopathic dilated cardiomyopathy (DCM) or ischemic cardiomyopathy (IC). However, the common or specific pathways of the initiation and processes of DCM and IC are still unclear. Here, we attempted to uncover the critical genes and potential molecular networks that play important roles in DCM and IC progression commonly or specifically. The transcriptional profiles from normal and DCM or IC patient samples were analyzed and compared using bioinformatic methods. Initially, the normal and DCM or IC sample data were processed and the most notable differentially expressed genes (DEGs) from DCM or IC were identified. By comparing the DEGs from DCM with those from IC, the DCM- and IC-specific DEGs were identified. The gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses indicated the significance of multiple biological processes as well as signaling pathways that affect heart function and DCM or IC progression. Protein-protein interaction network analysis identified the relationships between different genes, and some important genes such as MYC and FN1 were found to be hubs, which master each individual module of DCM-specific and IC-specific DEGs, respectively. We discovered commonalities and differences of gene expression profiles and molecular pathways between different cardiomyopathies. The gene discovery and molecular signature analysis in this study could offer insights into disease mechanisms and also identify markers useful for diagnostic, prognostic, and therapeutic purposes.
Assuntos
Cardiomiopatia Dilatada/genética , RNA/genética , Perfilação da Expressão Gênica/métodos , Ontologia Genética , Redes Reguladoras de Genes/genética , Humanos , Mapas de Interação de ProteínasRESUMO
We investigated the effect of Jianpi Bushen prescription (JBP) on the expression of the SHP-1 and apoptosis-related genes in chemically damaged model mice and a compound e-jiao slurry (EJS) group (positive control). Kunming mice received an abdominal injection of 100 mg/kg cyclophosphamide once a day for 3 consecutive days to induce chemical damage. The mice underwent lavage at a suspension of 0.1 g/kg low-dose JBP (100%), high-dose JBP (200%), and 0.2 mL/10 g EJS twice a day for 9 days. mRNA and protein expression of SHP-1 in bone marrow mononuclear cells was detected using real-time polymerase chain reaction and Western blot; mRNA expression of B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X (Bax) protein was detected by in situ hybridization. Expression of SHP-1 and Bax mRNA was significantly upregulated in the model group compared to the control group (P < 0.05). Expression in the low-dose JBP, high-dose JBP, and EJS groups was significantly downregulated compared with the model group (P < 0.05). The low-dose JBP group exhibited much lower SHP-1 and Bax mRNA expression levels. Compared with controls, Bcl-2 mRNA expression was significantly reduced in the model group (P < 0.05). Expression in the low-dose JBP, high-dose JBP, and EJS groups significantly increased compared with the model group (P < 0.05). The low-dose JBP group showed much higher Bcl-2 mRNA expression. Therefore, JBP regulates the expression of the SHP- 1, Bax, and Bcl-2 genes in chemically damaged mice.
Assuntos
Apoptose/genética , Medicamentos de Ervas Chinesas/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Proteína Tirosina Fosfatase não Receptora Tipo 6/genética , Animais , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/metabolismo , Ciclofosfamida/administração & dosagem , Ciclofosfamida/efeitos adversos , Regulação para Baixo , Medicamentos de Ervas Chinesas/administração & dosagem , Feminino , Masculino , Camundongos , Proteína Tirosina Fosfatase não Receptora Tipo 6/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , RNA Mensageiro/genética , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismoRESUMO
This study investigated the effect of the Jianpi Bushen Prescription (JBP) on the expression of 3 major proteins in chemically damaged model mice. The 3 proteins were the Wnt3a, the SHP-1, and the transcription factors (NF-E2, c-jun, and c-fos) of the AP-1 protein family. Kunming mice were randomly divided into chemically damaged group (N=48), which received an abdominal injection of (100 mg/kg) cyclophosphamide once a day for 3 consecutive days, and control group (N=12), which received the same amount of saline. Then, the chemically damaged mice were randomly divided into chemically damaged model group (N=12), which received 0.2 mL/10 g of saline twice a day for 9 days, positive control group (N=12), which received 0.2 mL/10 g of the e-jiao slurry (EJS) compound twice a day for 9 days, low dose JBP group (N=12), which received 0.1 g/kg suspension JBP (100% concentration) twice a day for 9 days and high dose JBP group (N=12), which received 0.1 g/kg suspension JBP (200% concentration) twice a day for 9 days. The bilateral femur and tibia bone marrow were collected from the mice in all groups. The protein expression of the specified proteins and transcription factors in the bone marrow mononuclear cells were detected by Western blot analysis. The results showed that the protein expression of Wnt3a was significantly downregulated in the chemically damaged model group compared to the control group (P<0.05). The low dose JBP, high dose JBP, and e-jiao slurry treatments significantly upregulated the protein expression of Wnt3a compared to the chemically damaged model group (P<0.05), with the low dose JBP producing the best results. Compared to the control group, the protein expressions of SHP-1, c-fos, c-jun, and NF-E2 were significantly higher in the chemically damaged model group (all P<0.05). The protein expressions of SHP-1, c-fos, c-jun, and NF-E2 were significantly lower in the chemically damaged model+the low dose JBP, chemically damaged model+high dose JBP, or chemically damaged model+EJS group compared to chemically damaged model (all P<0.05), with the low dose JBP producing the best results. These results indicate that JBP regulates the expressions of SHP-1, Wnt3a, and AP-1 proteins in chemically damaged mice.
Assuntos
Medicamentos de Ervas Chinesas/administração & dosagem , Proteína Tirosina Fosfatase não Receptora Tipo 6/biossíntese , Fator de Transcrição AP-1/biossíntese , Proteína Wnt3A/biossíntese , Animais , Células da Medula Óssea/efeitos dos fármacos , Células da Medula Óssea/metabolismo , Ciclofosfamida/toxicidade , Fêmur/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Camundongos , Tíbia/efeitos dos fármacosRESUMO
The effects of the traditional Chinese drug Jianpi Bushen Prescription (JBP) were investigated on expressions of Wnt3a and Cyclin D1 genes in radiation-damaged mice. The radiation damage model was induced in Kumming mice by single total body irradiation treatment for 9 days. Mice were divided into the radiation group, low-dose (100%) JBP group, high-dose (200%) JBP group, or batyl alcohol group (positive control), which were administered twice a day for 9 days. mRNA and protein expressions of Wnt3a were detected in bone marrow mononuclear cells by real-time polymerase chain reaction and Western blot, whereas Cyclin D1 mRNA was detected by in situ hybridization. Wnt3a expressions were significantly downregulated in the radiation damage model group compared with all other groups (P < 0.05). The positive cell rate of Cyclin D1 mRNA expression and the number of granulocyte macrophage colonies were significantly decreased in the radiation damage model group relative to all other groups (P < 0.05). Furthermore, mRNA and protein expressions of Wnt3a, the positive cell rate of Cyclin D1 mRNA expression in bone marrow cells, and the number of granulocyte macrophage colonies were all significantly higher in the low-dose JBP group than in the high-dose JBP group (P < 0.05). In summary, JBP plays a protective role on radiation-induced bone marrow through the activation of the Wnt3a signaling pathway, and promotes the transcription and expression of Cyclin D1.