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Preventive effects of Lactobacillus johnsonii on the renal injury of mice induced by high fluoride exposure: Insights from colonic microbiota and co-occurrence network analysis

Xin, Jinge, Sun, Ning, Wang, Hesong, Ma, Hailin, Wu, Bangyuan, Li, Lianxin, Wang, Yanyan, Huang, Haonan, Zeng, Dong, Bai, Xiuquan, Chen, Ali, Gong, Shenhai, Ni, Xueqin, Bai, Yang·2021·Ecotoxicology and Environmental Safety
DOI 10.1016/j.ecoenv.2021.113006
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Image 1
CC-BY-NC-ND · From the PDF
Fig. 1 Histopathological changes in the kidney tissue of mice. Histological observation of ileum under original magnification (100 × ). (A)Kideny tissue in Ctrl group; (B) Kideny tissue in F group; (C) Kideny tissue in Prob group. The swell, severe granular degeneration as well as vacuolization of the epithelial cells of fluorideexposure mice was indicated by white arrows.
Fig. 1 Histopathological changes in the kidney tissue of mice. Histological observation of ileum under original magnification (100 × ). (A)Kideny tissue in Ctrl group; (B) Kideny tissue in F group; (C) Kideny tissue in Prob group. The swell, severe granular degeneration as well as vacuolization of the epithelial cells of fluorideexposure mice was indicated by white arrows.CC-BY-NC-ND · From the PDF
Image 3
CC-BY-NC-ND · From the PDF
Fig. 3 Change of peroxidation damage and antioxidant capacity in the kidney. (A) MDA content. ( F 2, 27 = 37.37, P < 0.001). (B) PC content ( F 2, 27 = 53.38, P < 0.001). (C) ASA activity ( F 2, 27 = 4.52, P = 0.020). (D) AHR ability ( F 2, 27 = 14.83, P < 0.001). (E) SOD activity ( F 2, 27 = 25.38, P < 0.001). (F) CAT activity ( F 2, 27 = 7.85, P = 0.002). (G) GSH-Px activity ( F 2, 27 = 9.06, P = 0.001). (H) GSH content ( F 2, 27 = 12.33, P < 0.001). Data are presented with the mean ± standard deviation (n = 10). Significance between groups was indicated with asterisk(* p < 0.05, ** p < 0.01). MDA, malondialdehyde. PC: protein carbonyls. ASA: anti-su p eroxide anion; AHR: anti-hydroxyl radical. SOD, superoxide dismutase. CAT, catalase. GSH-Px, glutathione peroxidase. GSH, glutathione.
Fig. 3 Change of peroxidation damage and antioxidant capacity in the kidney. (A) MDA content. ( F 2, 27 = 37.37, P < 0.001). (B) PC content ( F 2, 27 = 53.38, P < 0.001). (C) ASA activity ( F 2, 27 = 4.52, P = 0.020). (D) AHR ability ( F 2, 27 = 14.83, P < 0.001). (E) SOD activity ( F 2, 27 = 25.38, P < 0.001). (F) CAT activity ( F 2, 27 = 7.85, P = 0.002). (G) GSH-Px activity ( F 2, 27 = 9.06, P = 0.001). (H) GSH content ( F 2, 27 = 12.33, P < 0.001). Data are presented with the mean ± standard deviation (n = 10). Significance between groups was indicated with asterisk(* p < 0.05, ** p < 0.01). MDA, malondialdehyde. PC: protein carbonyls. ASA: anti-su p eroxide anion; AHR: anti-hydroxyl radical. SOD, superoxide dismutase. CAT, catalase. GSH-Px, glutathione peroxidase. GSH, glutathione.CC-BY-NC-ND · From the PDF
Fig. 4 Linear Discriminant Analysis Effect Size Analysis. Significantly discriminative taxa (Biomarkers) between the Ctrl and F groups of mice were displayed by LDA value distribution histogram (A) and cladogram (C). Significantly discriminative taxa (Biomarkers) between the F and Prob groups of mice were displayed by LDA value distribution histogram (B) and cladogram (D). Only taxa meeting the LDA significance thresholds ( > 4) are shown. Different coloured bars and regions represent different groups. From the interior to the exterior, each layer represents the phylum, class, order, family, and genus levels.
Fig. 4 Linear Discriminant Analysis Effect Size Analysis. Significantly discriminative taxa (Biomarkers) between the Ctrl and F groups of mice were displayed by LDA value distribution histogram (A) and cladogram (C). Significantly discriminative taxa (Biomarkers) between the F and Prob groups of mice were displayed by LDA value distribution histogram (B) and cladogram (D). Only taxa meeting the LDA significance thresholds ( > 4) are shown. Different coloured bars and regions represent different groups. From the interior to the exterior, each layer represents the phylum, class, order, family, and genus levels.CC-BY-NC-ND · From the PDF
Fig. 5 Defining control specific and sensitive bacterial OTUs in colonic samples. (A) Bipartite networks of indicator species analysis. It display different treatment specific OTUs in the colonic bacterial communities using indicator species analysis. Circle represent individual bacteria and OTUs that are positively and significantly associated ( p < 0.05) with one or more of different grouping factor (association(s) given by connecting lines). OTUs are colored according to their Phylum assignment. (B) Different treatments are enriched and depleted for certain OTUs. Enrichment and depletion of bacterial OTUs included in differential abundance analysis for each treatments compared with other treatments were determined using DESeq2 analysis. Each point represents an individual OTU, and the position along the x axis and y axis represents the abundance fold change and p value, respectively, compared with other group. (C) Venn diagrams show the number of OTUs responding to different grouping factor identified with indicator species analysis (red) and by DESeq2 analysis (blue). OTUs identified by both methods were defined as control specific OTUs (csOTUs), high F exposure sensitive OTUs (fsOTUs) as well as high F exposure and L. johnsonii BS15 co-sensitive OTUs (flsOTUs).
Fig. 5 Defining control specific and sensitive bacterial OTUs in colonic samples. (A) Bipartite networks of indicator species analysis. It display different treatment specific OTUs in the colonic bacterial communities using indicator species analysis. Circle represent individual bacteria and OTUs that are positively and significantly associated ( p < 0.05) with one or more of different grouping factor (association(s) given by connecting lines). OTUs are colored according to their Phylum assignment. (B) Different treatments are enriched and depleted for certain OTUs. Enrichment and depletion of bacterial OTUs included in differential abundance analysis for each treatments compared with other treatments were determined using DESeq2 analysis. Each point represents an individual OTU, and the position along the x axis and y axis represents the abundance fold change and p value, respectively, compared with other group. (C) Venn diagrams show the number of OTUs responding to different grouping factor identified with indicator species analysis (red) and by DESeq2 analysis (blue). OTUs identified by both methods were defined as control specific OTUs (csOTUs), high F exposure sensitive OTUs (fsOTUs) as well as high F exposure and L. johnsonii BS15 co-sensitive OTUs (flsOTUs).CC-BY-NC-ND · From the PDF
Fig. 6 Characteristics of co-occurrence networks and sensitive OTUs. (A) Co-occurrence networks visualizing significant correlations ( ρ > 0.6, P < 0.01; indicated with gray lines) between OTU pairs in the Ctrl, F and Prob colonic bacterial communities. OTUs were colored by their association to the different representative meaning. Shaded areas represent the network modules containing csOTUs, flOTUs and flsOTUs. (B) Cumulative relative abundance of bacteria of different grouping sensitive modules in colonic network. The cumulative relative abundance in samples of csOTUs (red), flOTUs (blue) and flsOTUs (green) indicates the overall response of grouping sensitive modules to different treatments. (C) Qualitative taxonomic composition of grouping sensitive modules is reported as proportional OTUs numbers per phylum and compared to the overall taxonomic distribution in the entire dataset (column ' all ' ). (D) Defining network modules. Plot showing the number of OTUs in the top 10 most populated modules for the colonic co-occurrence networks. OTUs were colored by their association to the different grouping factor. Percentages on the X-axis indicate the proportion of csOTUs, flOTUs and flsOTUs present in each module. (E) Degree of co-occurrence and abundance of csOTUs, flOTUs and flsOTUs. Relative abundance of all OTUs from the colonic microbial co-occurence network was plotted as a function of their degree of cooccurrence. OTUs were colored by their association to the different grouping and keystone OTUs (top 3% degree of co-occurrence) have yellow background. Side panels recapitulate the distributions of co-occurence degrees and abundance for the csOTUs, flOTUs and flsOTUs (circle colored by association to different treat­ ments). Spearman ' s rank correlation between the keystone OTUs and (F) clinical biochemical parameters, (G) Oxidative stress and antioxidant damage, and (H) mRNA expression levels of colon tight junction proteins. (I) keystone OTUs that are significantly altered by excess fluoride intake compared with the control group and reverted by L. johnsonii BS15. The results were presented as heatmaps using R package pheatmap. Heatmap showed that partial microbial modules was intimately associated with renal and colonic functions, which was positively or negatively related to functional paramrter. Rho in the color key represented Spearman rank correlation coefficient. * P < 0.05 denoted statistical significance between the correlations of keystone OTUs and functional parameters.
Fig. 6 Characteristics of co-occurrence networks and sensitive OTUs. (A) Co-occurrence networks visualizing significant correlations ( ρ > 0.6, P < 0.01; indicated with gray lines) between OTU pairs in the Ctrl, F and Prob colonic bacterial communities. OTUs were colored by their association to the different representative meaning. Shaded areas represent the network modules containing csOTUs, flOTUs and flsOTUs. (B) Cumulative relative abundance of bacteria of different grouping sensitive modules in colonic network. The cumulative relative abundance in samples of csOTUs (red), flOTUs (blue) and flsOTUs (green) indicates the overall response of grouping sensitive modules to different treatments. (C) Qualitative taxonomic composition of grouping sensitive modules is reported as proportional OTUs numbers per phylum and compared to the overall taxonomic distribution in the entire dataset (column ' all ' ). (D) Defining network modules. Plot showing the number of OTUs in the top 10 most populated modules for the colonic co-occurrence networks. OTUs were colored by their association to the different grouping factor. Percentages on the X-axis indicate the proportion of csOTUs, flOTUs and flsOTUs present in each module. (E) Degree of co-occurrence and abundance of csOTUs, flOTUs and flsOTUs. Relative abundance of all OTUs from the colonic microbial co-occurence network was plotted as a function of their degree of cooccurrence. OTUs were colored by their association to the different grouping and keystone OTUs (top 3% degree of co-occurrence) have yellow background. Side panels recapitulate the distributions of co-occurence degrees and abundance for the csOTUs, flOTUs and flsOTUs (circle colored by association to different treat­ ments). Spearman ' s rank correlation between the keystone OTUs and (F) clinical biochemical parameters, (G) Oxidative stress and antioxidant damage, and (H) mRNA expression levels of colon tight junction proteins. (I) keystone OTUs that are significantly altered by excess fluoride intake compared with the control group and reverted by L. johnsonii BS15. The results were presented as heatmaps using R package pheatmap. Heatmap showed that partial microbial modules was intimately associated with renal and colonic functions, which was positively or negatively related to functional paramrter. Rho in the color key represented Spearman rank correlation coefficient. * P < 0.05 denoted statistical significance between the correlations of keystone OTUs and functional parameters.CC-BY-NC-ND · From the PDF

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Metadata

Authors
Xin, J., Sun, N., Wang, H., Ma, H., Wu, B., Li, L., Wang, Y., Huang, H., Zeng, D., Bai, X., Chen, A., Gong, S., Ni, X., & Bai, Y.
Venue
Ecotoxicology and Environmental Safety
Publisher
Elsevier BV
Published
2021-12
Volume
228
Pages
113006
ISSN
0147-6513
DOI
10.1016/j.ecoenv.2021.113006
Cited by
22
References
2
Language
en
Licence
Funders
  • Southern Medical University (2020Z008)
  • Sichuan Province Science and Technology Support Program (2021YFH0097)
  • Natural Science Foundation of Guangdong Province (2019A1515012115)

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