Evaluation of the mTORC activity in the presence of Toxoplasma gondii and azathioprine in human monocyte cell line
Abstract Background Autophagy is an important part of pathogenesis of IBD. Thiopurines such as azathioprine (AZA) are approved drugs for clinical practices in IBD patients. Besides, as an escape strategy, Toxoplasma gondii can use the mTORC1 complex to inactivate autophagy. Methods In this study, we...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-03-01
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| Series: | BMC Microbiology |
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| Online Access: | https://doi.org/10.1186/s12866-023-02819-8 |
| _version_ | 1797865422357266432 |
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| author | Sara Nemati Hanieh Mohammad Rahimi Anna Meyfour Hossein Pazoki Hamid Asadzadeh Aghdaei Shabnam Shahrokh Hamed Mirjalali |
| author_facet | Sara Nemati Hanieh Mohammad Rahimi Anna Meyfour Hossein Pazoki Hamid Asadzadeh Aghdaei Shabnam Shahrokh Hamed Mirjalali |
| author_sort | Sara Nemati |
| collection | DOAJ |
| description | Abstract Background Autophagy is an important part of pathogenesis of IBD. Thiopurines such as azathioprine (AZA) are approved drugs for clinical practices in IBD patients. Besides, as an escape strategy, Toxoplasma gondii can use the mTORC1 complex to inactivate autophagy. Methods In this study, we investigated whether T. gondii tachyzoites may modulate autophagy and interfere the effects of azathioprine in IBD treatment. PMA-activated human monocyte cell line (THP-1) was infected with fresh T. gondii RH tachyzoites. After 5 h of infection, the cells were treated with AZA for 6 h. The expression of atg5, atg7, atg12, lc3b, and β-actin (BACT) genes was evaluated using quantitative real-time PCR. To analyze the phosphorylation of ribosomal protein S6 (rpS6), western blot using specific primary antibodies was performed. Results The results of real-time PCR revealed that AZA, T. gondii tachyzoites, and a combination of AZA and T. gondii tachyzoites upregulated atg5 gene for 4.297-fold (P-value = 0.014), 2.49-fold (P-value = 0.006), and 4.76-fold (P-value = 0.001), respectively. The atg7 gene showed significant upregulation (2.272-fold; P-value = 0.014) and (1.51-fold; P-value = 0.020) in AZA and AZA / T. gondii, respectively. The expression of atg12 gene was significantly downregulated in AZA and T. gondii tachyzoites for (8.85-fold; P-value = 0.004) and (2.005-fold; P-value = 0.038), respectively, but upregulated in T. gondii/AZA (1.52-fold; P-value = 0.037). In addition, the lc3b gene was only significantly changed in AZA / T. gondii (3.028-fold; P-value = 0.001). Western blot analysis showed that T. gondii tachyzoites significantly phosphorylated rpS6, and tachyzoites did not interfere the effects of AZA to phosphorylate the rpS6. Conclusion Taken together, although AZA and T. gondii similarly affects the expression levels of atg5, atg7, and atg12, but T. gondii does not seem to modulate the effects of AZA via mTORC functions. |
| first_indexed | 2024-04-09T23:08:34Z |
| format | Article |
| id | doaj.art-95e31c65879a47dbba2db98d9be07dba |
| institution | Directory Open Access Journal |
| issn | 1471-2180 |
| language | English |
| last_indexed | 2024-04-09T23:08:34Z |
| publishDate | 2023-03-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Microbiology |
| spelling | doaj.art-95e31c65879a47dbba2db98d9be07dba2023-03-22T10:34:20ZengBMCBMC Microbiology1471-21802023-03-012311810.1186/s12866-023-02819-8Evaluation of the mTORC activity in the presence of Toxoplasma gondii and azathioprine in human monocyte cell lineSara Nemati0Hanieh Mohammad Rahimi1Anna Meyfour2Hossein Pazoki3Hamid Asadzadeh Aghdaei4Shabnam Shahrokh5Hamed Mirjalali6Foodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical SciencesFoodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical SciencesBasic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical SciencesDepartment of Medical Microbiology, Faculty of Medicine, Infectious Diseases Research Center, Gonabad University of Medical SciencesBasic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical SciencesGastroenterology and Liver Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical SciencesFoodborne and Waterborne Diseases Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical SciencesAbstract Background Autophagy is an important part of pathogenesis of IBD. Thiopurines such as azathioprine (AZA) are approved drugs for clinical practices in IBD patients. Besides, as an escape strategy, Toxoplasma gondii can use the mTORC1 complex to inactivate autophagy. Methods In this study, we investigated whether T. gondii tachyzoites may modulate autophagy and interfere the effects of azathioprine in IBD treatment. PMA-activated human monocyte cell line (THP-1) was infected with fresh T. gondii RH tachyzoites. After 5 h of infection, the cells were treated with AZA for 6 h. The expression of atg5, atg7, atg12, lc3b, and β-actin (BACT) genes was evaluated using quantitative real-time PCR. To analyze the phosphorylation of ribosomal protein S6 (rpS6), western blot using specific primary antibodies was performed. Results The results of real-time PCR revealed that AZA, T. gondii tachyzoites, and a combination of AZA and T. gondii tachyzoites upregulated atg5 gene for 4.297-fold (P-value = 0.014), 2.49-fold (P-value = 0.006), and 4.76-fold (P-value = 0.001), respectively. The atg7 gene showed significant upregulation (2.272-fold; P-value = 0.014) and (1.51-fold; P-value = 0.020) in AZA and AZA / T. gondii, respectively. The expression of atg12 gene was significantly downregulated in AZA and T. gondii tachyzoites for (8.85-fold; P-value = 0.004) and (2.005-fold; P-value = 0.038), respectively, but upregulated in T. gondii/AZA (1.52-fold; P-value = 0.037). In addition, the lc3b gene was only significantly changed in AZA / T. gondii (3.028-fold; P-value = 0.001). Western blot analysis showed that T. gondii tachyzoites significantly phosphorylated rpS6, and tachyzoites did not interfere the effects of AZA to phosphorylate the rpS6. Conclusion Taken together, although AZA and T. gondii similarly affects the expression levels of atg5, atg7, and atg12, but T. gondii does not seem to modulate the effects of AZA via mTORC functions.https://doi.org/10.1186/s12866-023-02819-8Toxoplasma gondiiAutophagymTORCAzathioprineInflammatory Bowel Diseases |
| spellingShingle | Sara Nemati Hanieh Mohammad Rahimi Anna Meyfour Hossein Pazoki Hamid Asadzadeh Aghdaei Shabnam Shahrokh Hamed Mirjalali Evaluation of the mTORC activity in the presence of Toxoplasma gondii and azathioprine in human monocyte cell line BMC Microbiology Toxoplasma gondii Autophagy mTORC Azathioprine Inflammatory Bowel Diseases |
| title | Evaluation of the mTORC activity in the presence of Toxoplasma gondii and azathioprine in human monocyte cell line |
| title_full | Evaluation of the mTORC activity in the presence of Toxoplasma gondii and azathioprine in human monocyte cell line |
| title_fullStr | Evaluation of the mTORC activity in the presence of Toxoplasma gondii and azathioprine in human monocyte cell line |
| title_full_unstemmed | Evaluation of the mTORC activity in the presence of Toxoplasma gondii and azathioprine in human monocyte cell line |
| title_short | Evaluation of the mTORC activity in the presence of Toxoplasma gondii and azathioprine in human monocyte cell line |
| title_sort | evaluation of the mtorc activity in the presence of toxoplasma gondii and azathioprine in human monocyte cell line |
| topic | Toxoplasma gondii Autophagy mTORC Azathioprine Inflammatory Bowel Diseases |
| url | https://doi.org/10.1186/s12866-023-02819-8 |
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