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Wei-Hua Huang
Central Couth University
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Background: Ginsenoside CK (GCK) serves as the potential anti-colorectal cancer (CRC) protopanaxadiol (PPD)-type saponin, which could be mainly bio-converted to yield PPD by gut microbiota. Meanwhile, the anti-CRC effects of GCK could be altered by gut microbiota due to its different diversity in CRC patients. We aimed to investigate the bioconversion variation of GCK mediated by gut microbiota from CRC patients by comparing with healthy subjects.
Methods: Gut microbiota profiled by 16S rRNA gene sequencing was collected from healthy volunteers and CRC patients. GCK was incubated with gut microbiota in vitro. A LC-MS/MS method was validated to quantify GCK and PPD after incubation at different time points.
Results: The bioconversion of GCK in healthy subjects group was much faster than CRC group, as well as the yield of PPD. Moreover, significant difference of PPD concentration between healthy subjects group and CRC group could be observed at 12 h, 48 h and 72 h check points. According to 16S rRNA sequencing, the profiles of gut microbiota derived from healthy volunteers and CRC patients significantly varied, in which 12 differentially abundant taxon were found, such as Bifidobacterium, Roseburia, Bacteroides and Collinsella. Spearman’s correlation analysis showed bacteria enriched in healthy subjects group were positively associated with biotransformation of GCK, while bacteria enriched in CRC group displayed non correlation characters. Among them, Roseburia which could secrete β-glycosidase showed the strongest positive association with the bioconversion of GCK.
Conclusion: The bioconversion of GCK in healthy subjects was much faster than CRC patients mediated by gut microbiota, which might alter the anti-CRC effects of GCK.
Keywords
Ginsenoside CK, gut microbiota, LC-MS/MS, colorectal cancer, 16S rRNA gene sequencing
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This is a list of supplementary files associated with this preprint. Click to download.
- ChineseMedicineHWH01292021GraphicAbstract.pptx
- ChineseMedicineHWH01292021SupplementaryFigure.pptx
SFigure 1: Representative MRM chromatograms of blank gut microbiota solution, GCK, PPD and PPT. a1,b1,c1 were the corresponding chromatograms of blank gut microbiota solution; a2: GCK; b2: PPD; c2: PPT. SFigure 2: Relative abundance of differentially abundant bacteria in each sample. (a) Bifidobacterium; (b) Blautia; (c) Corynebacterium; (d) Enhydrobacter; (e) Rothia; (f) SMB53; (g) Bacteroides; (h) Collinsella; (i) Coprobacillus; (j) Enterobacteriaceae; significant difference of relative abundance were analyzed using Kruskal-Wallis test; * p<0.05, ** p<0.01 and # p<0.001. SFigure3: Significant functional pathways between health and CRC group by PICRUSt analysis (p<0.05, LDA scores >2).
- ChineseMedicineHWH01292021SupplementaryTable.docx
Supplementary Table 1: MRM parameters of GCK, PPD and PPT Supplementary Table 2: Linearity range, correlation coefficients (r), calibration curves and LLOQ of GCK and PPD Supplementary Table 3: Precision and accuracy of GCK and PPD Supplementary Table 4: Recovery and matrix effect of GCK and PPD Supplementary Table 5: Taxonomical information of the differentially abundance bacteria by LEfSe analysis
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View the published article at Chinese Medicine.
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On 10 Feb, 2021
Reviewer #1 agreed
On 09 Feb, 2021
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Received 09 Feb, 2021
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A new preprint design is coming!
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See the published version of this preprint at Chinese Medicine.
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
Wei-Hua Huang
Central Couth University
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 Chinese Medicine.
Version 1
Posted 05 Feb, 2021
No community comments so far
Editorial decision: Major revision
On 23 Feb, 2021
Review #3 received
Received 22 Feb, 2021
Review #2 received
Received 21 Feb, 2021
Reviewer #3 agreed
On 10 Feb, 2021
Reviewer #2 agreed
On 10 Feb, 2021
Reviewer #1 agreed
On 09 Feb, 2021
Review #1 received
Received 09 Feb, 2021
Reviews received
Received 09 Feb, 2021
Reviewers invited
Invitations sent on 07 Feb, 2021
Editor assigned
On 29 Jan, 2021
Submission checks complete
On 29 Jan, 2021
Editor invited
On 29 Jan, 2021
First submitted
On 28 Jan, 2021
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Internal Medicine
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Chinese Medicine.
Background: Ginsenoside CK (GCK) serves as the potential anti-colorectal cancer (CRC) protopanaxadiol (PPD)-type saponin, which could be mainly bio-converted to yield PPD by gut microbiota. Meanwhile, the anti-CRC effects of GCK could be altered by gut microbiota due to its different diversity in CRC patients. We aimed to investigate the bioconversion variation of GCK mediated by gut microbiota from CRC patients by comparing with healthy subjects.
Methods: Gut microbiota profiled by 16S rRNA gene sequencing was collected from healthy volunteers and CRC patients. GCK was incubated with gut microbiota in vitro. A LC-MS/MS method was validated to quantify GCK and PPD after incubation at different time points.
Results: The bioconversion of GCK in healthy subjects group was much faster than CRC group, as well as the yield of PPD. Moreover, significant difference of PPD concentration between healthy subjects group and CRC group could be observed at 12 h, 48 h and 72 h check points. According to 16S rRNA sequencing, the profiles of gut microbiota derived from healthy volunteers and CRC patients significantly varied, in which 12 differentially abundant taxon were found, such as Bifidobacterium, Roseburia, Bacteroides and Collinsella. Spearman’s correlation analysis showed bacteria enriched in healthy subjects group were positively associated with biotransformation of GCK, while bacteria enriched in CRC group displayed non correlation characters. Among them, Roseburia which could secrete β-glycosidase showed the strongest positive association with the bioconversion of GCK.
Conclusion: The bioconversion of GCK in healthy subjects was much faster than CRC patients mediated by gut microbiota, which might alter the anti-CRC effects of GCK.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
This is a list of supplementary files associated with this preprint. Click to download.
- ChineseMedicineHWH01292021GraphicAbstract.pptx
- ChineseMedicineHWH01292021SupplementaryFigure.pptx
SFigure 1: Representative MRM chromatograms of blank gut microbiota solution, GCK, PPD and PPT. a1,b1,c1 were the corresponding chromatograms of blank gut microbiota solution; a2: GCK; b2: PPD; c2: PPT. SFigure 2: Relative abundance of differentially abundant bacteria in each sample. (a) Bifidobacterium; (b) Blautia; (c) Corynebacterium; (d) Enhydrobacter; (e) Rothia; (f) SMB53; (g) Bacteroides; (h) Collinsella; (i) Coprobacillus; (j) Enterobacteriaceae; significant difference of relative abundance were analyzed using Kruskal-Wallis test; * p<0.05, ** p<0.01 and # p<0.001. SFigure3: Significant functional pathways between health and CRC group by PICRUSt analysis (p<0.05, LDA scores >2).
- ChineseMedicineHWH01292021SupplementaryTable.docx
Supplementary Table 1: MRM parameters of GCK, PPD and PPT Supplementary Table 2: Linearity range, correlation coefficients (r), calibration curves and LLOQ of GCK and PPD Supplementary Table 3: Precision and accuracy of GCK and PPD Supplementary Table 4: Recovery and matrix effect of GCK and PPD Supplementary Table 5: Taxonomical information of the differentially abundance bacteria by LEfSe analysis
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