Interference of pseudorabies virus infection on functions of porcine granulosa cells via apoptosis modulated by MAPK signaling pathways
Abstract Background Pseudorabies virus (PRV) is one of the major viral pathogens leading to reproductive disorders in swine. However, little is known about the effects of PRV infection on porcine reproductive system. Ovarian granulosa cells are somatic cells surrounding oocytes in ovary and required...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2024-01-01
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| Series: | Virology Journal |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12985-024-02289-y |
| _version_ | 1797340311076208640 |
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| author | Lingcong Deng Wenpeng Min Songyangnian Guo Jiping Deng Xiaosong Wu Dewen Tong Anwen Yuan Qing Yang |
| author_facet | Lingcong Deng Wenpeng Min Songyangnian Guo Jiping Deng Xiaosong Wu Dewen Tong Anwen Yuan Qing Yang |
| author_sort | Lingcong Deng |
| collection | DOAJ |
| description | Abstract Background Pseudorabies virus (PRV) is one of the major viral pathogens leading to reproductive disorders in swine. However, little is known about the effects of PRV infection on porcine reproductive system. Ovarian granulosa cells are somatic cells surrounding oocytes in ovary and required for folliculogenesis. The present study aimed to investigate the interference of PRV on functions of porcine ovarian granulosa cells in vitro. Methods Primary granulosa cells were isolated from porcine ovaries. To investigate the PRV infectivity, transmission electron microscopy (TEM) was used to check the presence of viral particles, and the expression of viral gE gene was detected by quantitative real-time PCR (qPCR) in PRV-inoculated cells. After PRV infection, cell viability was detected by MTS assay, Ki67 for proliferative status was determined by immunofluorescence assay (IFA), cell cycle and apoptosis were detected by flow cytometry, and progesterone (P4) and estradiol (E2) were determined by radioimmunoassay. The checkpoint genes of cell cycle and apoptosis-related proteins were studied by qPCR and western blotting. Results Virus particles were observed in the nucleus and cytoplasm of PRV-infected granulosa cells by TEM imaging, and the expression of viral gE gene increased in a time-dependent manner post infection. PRV infection inhibited cell viability and blocked cell cycle at S phase in porcine granulosa cells, accompanied by decreases in expression of Ki67 protein and checkpoint genes related to S phase. Radioimmunoassay revealed decreased levels in P4 and E2, and the expressions of key steroidogenic enzymes were also down-regulated post PRV-infection. In addition, PRV induced apoptosis with an increase in Bax expression and activation of caspase 9, and the phosphorylation of JNK, ERK and p38 MAPKs were significantly up-regulated in porcine ovarian granulosa cells post PRV infection. Conclusions The data indicate that PRV causes infection on porcine ovarian granulosa cells and interferes the cell functions through apoptosis, and the MAPK signaling pathway is involved in the viral pathogenesis. |
| first_indexed | 2024-03-08T10:01:10Z |
| format | Article |
| id | doaj.art-cbeb4df793c24edba56da62187721c74 |
| institution | Directory Open Access Journal |
| issn | 1743-422X |
| language | English |
| last_indexed | 2024-03-08T10:01:10Z |
| publishDate | 2024-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Virology Journal |
| spelling | doaj.art-cbeb4df793c24edba56da62187721c742024-01-29T10:57:38ZengBMCVirology Journal1743-422X2024-01-0121111310.1186/s12985-024-02289-yInterference of pseudorabies virus infection on functions of porcine granulosa cells via apoptosis modulated by MAPK signaling pathwaysLingcong Deng0Wenpeng Min1Songyangnian Guo2Jiping Deng3Xiaosong Wu4Dewen Tong5Anwen Yuan6Qing Yang7College of Veterinary Medicine, Hunan Agricultural UniversityCollege of Veterinary Medicine, Hunan Agricultural UniversityCollege of Veterinary Medicine, Hunan Agricultural UniversityCollege of Veterinary Medicine, Hunan Agricultural UniversityCollege of Veterinary Medicine, Hunan Agricultural UniversityCollege of Veterinary Medicine, Northwest A&F UniversityCollege of Veterinary Medicine, Hunan Agricultural UniversityCollege of Veterinary Medicine, Hunan Agricultural UniversityAbstract Background Pseudorabies virus (PRV) is one of the major viral pathogens leading to reproductive disorders in swine. However, little is known about the effects of PRV infection on porcine reproductive system. Ovarian granulosa cells are somatic cells surrounding oocytes in ovary and required for folliculogenesis. The present study aimed to investigate the interference of PRV on functions of porcine ovarian granulosa cells in vitro. Methods Primary granulosa cells were isolated from porcine ovaries. To investigate the PRV infectivity, transmission electron microscopy (TEM) was used to check the presence of viral particles, and the expression of viral gE gene was detected by quantitative real-time PCR (qPCR) in PRV-inoculated cells. After PRV infection, cell viability was detected by MTS assay, Ki67 for proliferative status was determined by immunofluorescence assay (IFA), cell cycle and apoptosis were detected by flow cytometry, and progesterone (P4) and estradiol (E2) were determined by radioimmunoassay. The checkpoint genes of cell cycle and apoptosis-related proteins were studied by qPCR and western blotting. Results Virus particles were observed in the nucleus and cytoplasm of PRV-infected granulosa cells by TEM imaging, and the expression of viral gE gene increased in a time-dependent manner post infection. PRV infection inhibited cell viability and blocked cell cycle at S phase in porcine granulosa cells, accompanied by decreases in expression of Ki67 protein and checkpoint genes related to S phase. Radioimmunoassay revealed decreased levels in P4 and E2, and the expressions of key steroidogenic enzymes were also down-regulated post PRV-infection. In addition, PRV induced apoptosis with an increase in Bax expression and activation of caspase 9, and the phosphorylation of JNK, ERK and p38 MAPKs were significantly up-regulated in porcine ovarian granulosa cells post PRV infection. Conclusions The data indicate that PRV causes infection on porcine ovarian granulosa cells and interferes the cell functions through apoptosis, and the MAPK signaling pathway is involved in the viral pathogenesis.https://doi.org/10.1186/s12985-024-02289-yPseudorabies virusOvarian granulosa cellsSteroidogenesisApoptosisMAPK signaling pathway |
| spellingShingle | Lingcong Deng Wenpeng Min Songyangnian Guo Jiping Deng Xiaosong Wu Dewen Tong Anwen Yuan Qing Yang Interference of pseudorabies virus infection on functions of porcine granulosa cells via apoptosis modulated by MAPK signaling pathways Virology Journal Pseudorabies virus Ovarian granulosa cells Steroidogenesis Apoptosis MAPK signaling pathway |
| title | Interference of pseudorabies virus infection on functions of porcine granulosa cells via apoptosis modulated by MAPK signaling pathways |
| title_full | Interference of pseudorabies virus infection on functions of porcine granulosa cells via apoptosis modulated by MAPK signaling pathways |
| title_fullStr | Interference of pseudorabies virus infection on functions of porcine granulosa cells via apoptosis modulated by MAPK signaling pathways |
| title_full_unstemmed | Interference of pseudorabies virus infection on functions of porcine granulosa cells via apoptosis modulated by MAPK signaling pathways |
| title_short | Interference of pseudorabies virus infection on functions of porcine granulosa cells via apoptosis modulated by MAPK signaling pathways |
| title_sort | interference of pseudorabies virus infection on functions of porcine granulosa cells via apoptosis modulated by mapk signaling pathways |
| topic | Pseudorabies virus Ovarian granulosa cells Steroidogenesis Apoptosis MAPK signaling pathway |
| url | https://doi.org/10.1186/s12985-024-02289-y |
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