See the published version of this preprint at BMC Microbiology.
This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research article
Yanmin Xu
Hongqi Hospital
Corresponding Author
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: Clostridioides (formerly Clostridium) difficile infection is the leading cause of antibiotic-associated colitis. Studies have demonstrated that C. difficile toxin A (TcdA) can cause apoptosis in many human cell types. The purpose of this study was to investigate the relationships among exposure to TcdA, the role of globular heads receptor of C1q (gC1qR) gene and the underlying intracellular apoptotic mechanism of human colonic epithelial cells (NCM 460).
Methods: In this study, gC1qR expression was examined using real-time polymerase chain reaction (PCR), western blot and immunohistochemical staining analysis. Cells viability was assessed by the water-soluble tetrazolium salt (WST-1) assay, apoptosis of cells was assessed by flow cytometry and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) Assay. Mitochondrial function was assessed via reactive oxygen species (ROS) generation, changes in the mitochondrial membrane potential (ΔΨm) and the content of ATP.
Results: Our study demonstrated that the concentration of TcdA increasing from 10 ng/ml to 20 ng/ml inhibited cell viability and induced cell apoptosis ( p < 0.01). Moreover, TcdA induced gC1qR mRNA and protein expression. Overexpression of gC1qR could cause the mitochondrial dysfunction (including production of ROS, decrease of ΔΨm and the content of ATP) and cell apoptosis. However, silencing of gC1qR gene could reverse the TcdA-induced cell apoptosis and mitochondrial dysfunction.
Conclusion: Therefore, these data support a mechanism wherein gC1qR plays a crucial role in TcdA-induced human colonic epithelial cells apoptosis involving a mitochondrial dependent manner.
Keywords
Clostridium difficile, TcdA, gC1qR, Mitochondrial function
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
This is a list of supplementary files associated with this preprint. Click to download.
Loading...
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
View the published article at BMC Microbiology.
Version 1
Posted 11 Mar, 2020
No community comments so far
Editorial decision: Major revision
On 23 Apr, 2020
Review #3 received
Received 21 Apr, 2020
Reviewer #3 agreed
On 09 Apr, 2020
Review #2 received
Received 25 Mar, 2020
Review #1 received
Received 23 Mar, 2020
Reviewer #2 agreed
On 19 Mar, 2020
Reviewer #1 agreed
On 19 Mar, 2020
Reviewers invited
Invitations sent on 11 Mar, 2020
Submission checks complete
On 06 Mar, 2020
Editor assigned
On 06 Mar, 2020
Editor invited
On 05 Mar, 2020
First submitted
On 04 Mar, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Learn more about our company.
See the published version of this preprint at BMC Microbiology.
This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Research article
Yanmin Xu
Hongqi Hospital
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
View the published article at BMC Microbiology.
Version 1
Posted 11 Mar, 2020
No community comments so far
Editorial decision: Major revision
On 23 Apr, 2020
Review #3 received
Received 21 Apr, 2020
Reviewer #3 agreed
On 09 Apr, 2020
Review #2 received
Received 25 Mar, 2020
Review #1 received
Received 23 Mar, 2020
Reviewer #2 agreed
On 19 Mar, 2020
Reviewer #1 agreed
On 19 Mar, 2020
Reviewers invited
Invitations sent on 11 Mar, 2020
Submission checks complete
On 06 Mar, 2020
Editor assigned
On 06 Mar, 2020
Editor invited
On 05 Mar, 2020
First submitted
On 04 Mar, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at BMC Microbiology.
Background: Clostridioides (formerly Clostridium) difficile infection is the leading cause of antibiotic-associated colitis. Studies have demonstrated that C. difficile toxin A (TcdA) can cause apoptosis in many human cell types. The purpose of this study was to investigate the relationships among exposure to TcdA, the role of globular heads receptor of C1q (gC1qR) gene and the underlying intracellular apoptotic mechanism of human colonic epithelial cells (NCM 460).
Methods: In this study, gC1qR expression was examined using real-time polymerase chain reaction (PCR), western blot and immunohistochemical staining analysis. Cells viability was assessed by the water-soluble tetrazolium salt (WST-1) assay, apoptosis of cells was assessed by flow cytometry and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) Assay. Mitochondrial function was assessed via reactive oxygen species (ROS) generation, changes in the mitochondrial membrane potential (ΔΨm) and the content of ATP.
Results: Our study demonstrated that the concentration of TcdA increasing from 10 ng/ml to 20 ng/ml inhibited cell viability and induced cell apoptosis ( p < 0.01). Moreover, TcdA induced gC1qR mRNA and protein expression. Overexpression of gC1qR could cause the mitochondrial dysfunction (including production of ROS, decrease of ΔΨm and the content of ATP) and cell apoptosis. However, silencing of gC1qR gene could reverse the TcdA-induced cell apoptosis and mitochondrial dysfunction.
Conclusion: Therefore, these data support a mechanism wherein gC1qR plays a crucial role in TcdA-induced human colonic epithelial cells apoptosis involving a mitochondrial dependent manner.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Loading...