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Exploration of Anti-inflammatory Mechanism of Forsythiaside A and Forsythiaside B in CuSO4-Induced Inflammation in Zebrafish by Metabolomic and Proteomic Analyses
Yunxia Li
Corresponding Author
ORCiD: https://orcid.org/0000-0003-1307-4335
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: Inflammation is a general pathological phenomenon during severe disturbances to the homeostasis. Forsythiaside A (FA) and Forsythiaside B (FB), isolated from the dried fruit of Forsythia suspensa (Thunb.) Vahl, are phenylethanoid compounds that show a significant anti-inflammatory effect. However, the properties and therapeutic mechanisms of this effect have not yet been systematically elucidated.
Methods: In this study, the anti-inflammatory effects of FA and FB were investigated in CuSO4-induced inflammation in zebrafish larvae. Intracellular generation of reactive oxygen species (ROS) and nitric oxide (NO) was investigated using fluorescence probes. Metabolomic and proteomic analyses using liquid chromatography-mass spectrometry were carried out to identify the expressions of metabolites and proteins associated with the anti-inflammatory mechanism of FA and FB. Quantitative polymerase chain reaction (PCR) was performed to detect the progressive changes in gene expression.
Results: FA and FB inhibited neutrophils migration to the damaged neuromasts and remarkably reduced CuSO4-induced ROS and NO generation in zebrafish larvae. Metabolomic analysis pointed to the involvement of Nicotinate and nicotinamide metabolism, Energy metabolism, Pyrimidine metabolism, and Purine metabolism. Proteomic analysis identified 146 differentially-expressed proteins between the control and model groups. These included Collagen [collagen type II alpha 1b precursor (col2a1b), collagen alpha-2(IX) chain precursor (col9a2), collagen type IX alpha I precursor (col9a1b)], Nucleoside diphosphate kinase 3 isoform X1 (Nme3), WD repeat-containing protein 3 (Wdr3), and 28S ribosomal protein S7 mitochondrial precursor (Mrps7). FA and FB were shown to reverse the abnormal expressions of potential metabolite and protein biomarkers and alleviate CuSO4-induced damage to the neuromasts in the zebrafish lateral line.
Conclusions: Our results indicate that FA and FB possess remarkable anti-inflammatory properties, protecting against CuSO4-induced neuromasts damage in zebrafish larvae. The results also suggest a multi-component and multi-regulatory therapeutic mechanism for FA and FB.
Keywords
Forsythiaside A, Forsythiaside B, Zebrafish, Inflammation, Metabolomics, Proteomics
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Received 29 Apr, 2020
Review #1 received
Received 28 Apr, 2020
Reviewer #2 agreed
On 21 Apr, 2020
Reviewers invited
Invitations sent on 18 Apr, 2020
Reviewer #1 agreed
On 18 Apr, 2020
Editor assigned
On 16 Apr, 2020
Submission checks complete
On 15 Apr, 2020
Editor invited
On 15 Apr, 2020
No comments provided
Editorial decision: Major Revision
On 17 Mar, 2020
Review #2 received
Received 12 Mar, 2020
Reviewer #2 agreed
On 12 Feb, 2020
Review #1 received
Received 15 Jan, 2020
Reviewer #1 agreed
On 02 Jan, 2020
Reviewers invited
Invitations sent on 23 Dec, 2019
First submitted
On 22 Dec, 2019
Editor assigned
On 22 Dec, 2019
Submission checks complete
On 21 Dec, 2019
Editor invited
On 21 Dec, 2019
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Subject Areas
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This preprint is under consideration at Journal of Neuroinflammation. A preprint is 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
Exploration of Anti-inflammatory Mechanism of Forsythiaside A and Forsythiaside B in CuSO4-Induced Inflammation in Zebrafish by Metabolomic and Proteomic Analyses
Yunxia Li
Corresponding Author
ORCiD: https://orcid.org/0000-0003-1307-4335
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: Under Review
Version 3
Posted 01 Jun, 2020
No community comments so far
Editor assigned
On 21 May, 2020
Submission checks complete
On 20 May, 2020
Editor invited
On 20 May, 2020
No comments provided
Editorial decision: Minor Revision
On 09 May, 2020
Review #2 received
Received 29 Apr, 2020
Review #1 received
Received 28 Apr, 2020
Reviewer #2 agreed
On 21 Apr, 2020
Reviewers invited
Invitations sent on 18 Apr, 2020
Reviewer #1 agreed
On 18 Apr, 2020
Editor assigned
On 16 Apr, 2020
Submission checks complete
On 15 Apr, 2020
Editor invited
On 15 Apr, 2020
No comments provided
Editorial decision: Major Revision
On 17 Mar, 2020
Review #2 received
Received 12 Mar, 2020
Reviewer #2 agreed
On 12 Feb, 2020
Review #1 received
Received 15 Jan, 2020
Reviewer #1 agreed
On 02 Jan, 2020
Reviewers invited
Invitations sent on 23 Dec, 2019
First submitted
On 22 Dec, 2019
Editor assigned
On 22 Dec, 2019
Submission checks complete
On 21 Dec, 2019
Editor invited
On 21 Dec, 2019
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Neurobiology of Disease
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Background: Inflammation is a general pathological phenomenon during severe disturbances to the homeostasis. Forsythiaside A (FA) and Forsythiaside B (FB), isolated from the dried fruit of Forsythia suspensa (Thunb.) Vahl, are phenylethanoid compounds that show a significant anti-inflammatory effect. However, the properties and therapeutic mechanisms of this effect have not yet been systematically elucidated.
Methods: In this study, the anti-inflammatory effects of FA and FB were investigated in CuSO4-induced inflammation in zebrafish larvae. Intracellular generation of reactive oxygen species (ROS) and nitric oxide (NO) was investigated using fluorescence probes. Metabolomic and proteomic analyses using liquid chromatography-mass spectrometry were carried out to identify the expressions of metabolites and proteins associated with the anti-inflammatory mechanism of FA and FB. Quantitative polymerase chain reaction (PCR) was performed to detect the progressive changes in gene expression.
Results: FA and FB inhibited neutrophils migration to the damaged neuromasts and remarkably reduced CuSO4-induced ROS and NO generation in zebrafish larvae. Metabolomic analysis pointed to the involvement of Nicotinate and nicotinamide metabolism, Energy metabolism, Pyrimidine metabolism, and Purine metabolism. Proteomic analysis identified 146 differentially-expressed proteins between the control and model groups. These included Collagen [collagen type II alpha 1b precursor (col2a1b), collagen alpha-2(IX) chain precursor (col9a2), collagen type IX alpha I precursor (col9a1b)], Nucleoside diphosphate kinase 3 isoform X1 (Nme3), WD repeat-containing protein 3 (Wdr3), and 28S ribosomal protein S7 mitochondrial precursor (Mrps7). FA and FB were shown to reverse the abnormal expressions of potential metabolite and protein biomarkers and alleviate CuSO4-induced damage to the neuromasts in the zebrafish lateral line.
Conclusions: Our results indicate that FA and FB possess remarkable anti-inflammatory properties, protecting against CuSO4-induced neuromasts damage in zebrafish larvae. The results also suggest a multi-component and multi-regulatory therapeutic mechanism for FA and FB.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8