Article Figures & Data
Figures
- Figure 1.
Schematic representation of the animal study design. A, long-term study (21-week after azoxymethane follow-up). B, short-term study (2- and 4-week after azoxymethane follow-up). Solid arrows indicate azoxymethane injection (10 mg/kg body weight, weekly i.p.) and empty arrows indicate saline i.p. injection.
- Figure 2.
Effect of SJW diet on overall survival, rectal bleeding, and biochemical parameters in azoxymethane-treated mice. A, comparison of the Kaplan-Meier survival curves for saline-treated mice on control (Con/Sal; black line) or 5% SJW diet (SJW/Sal; green line), azoxymethane-treated mice on control diet (Con/AOM; red line), 2.5% (SJW(L)/AOM, orange line) or 5% SJW diet (SJW/AOM, blue line). The median survival for each group is mentioned on the right side. P values for difference in survival were calculated using the Mantel–Cox (log-rank) test. B, rectal bleeding score of mice under study on a relative scale of 0 to 10. C, body weight gain of mice during the course of the study with respect to initial body weight. Change in (D) serum albumin, (E) BUN, and (F) cholesterol levels in mice. The statistical analysis for B–F was analyzed using one-way ANOVA with the Tukey correction for multiple testing. Color codes are same as those used for A. *, P < 0.05; **, P < 0.01; ***, P < 0.001; and ****, P < 0.0001, respectively. NS, not significant.
- Figure 3.
Effect of SJW diet on azoxymethane-induced colorectal tumorigenesis. Scatter plots for (A) total number of tumors (multiplicity), (B) number of tumors with diameter > 2 mm and (C) total tumor volume in azoxymethane-treated mice on control diet (red) or 2.5% (orange) or 5% (blue) SJW- diet. Scatter plots for number of polyps found in the colon of azoxymethane-treated mice on control (red box) or 5% SJW diet (blue triangle) after (D) 2 week or (E) 4 week of last azoxymethane injection. Statistical significance was calculated using one-way ANOVA with the Tukey correction for multiple testing for A, B, and C. The two-tailed Mann–Whitney test was performed for D and E. *, P < 0.05; **, P < 0.01; ***, P < 0.001; and ****, P < 0.0001, respectively.
- Figure 4.
Pathways found to be downregulated in the colon epithelium of azoxymethane-treated mice due to SJW diet. Microarray-based analysis of gene expression and the Ingenuity Pathway Analysis revealed downregulation of (A) NF-κB (A) and, ERK1/2 signaling (B). C, heatmap showing relative expression level of genes related to NF-κB and ERK1/2 signaling pathways in normal colon epithelium from saline-treated mice on control diet (Con/Sal) or 5% SJW diet (SJW/Sal) and non-tumor colon epithelium from azoxymethane-treated mice on control diet (Con/AOM) or 5% SJW diet (SJW/AOM).
- Figure 5.
qRT-PCR analysis of changes in the expression of representative genes related to NF-κB and ERK1/2 signaling pathway in colon epithelium due to SJW diet. All values are presented as mean + standard deviation. Statistical significance was calculated using one-way ANOVA with the Tukey correction for multiple testing *, P < 0.05; **, P < 0.01; ***, P < 0.001; and ****, P < 0.0001, respectively. NS, not significant.
- Figure 6.
qRT-PCR analysis of changes in the expression of representative genes related to NF-κB and ERK1/2 signaling pathway in colon epithelium due to SJW diet after just a 2 weeks (A) or 4 weeks (B) following the last azoxymethane injection: short-term study. Statistical significance was calculated using one-way ANOVA with the Tukey correction for multiple testing. *, P < 0.05; **, P < 0.01; ***, P < 0.001; and ****, P < 0.0001, respectively. NS, not significant.
Additional Files
Supplementary Data
Files in this Data Supplement:
- Supplementary Materials and Methods - Extraction of constituents from the SJW Diet and SJW Powder UPLC-MS Analysis
- Supplementary Figure Legends - Supplementary Figure Legends
- Supplementary Table S1 - Supplementary Table S1. Composition of St. John's wort extract.
- Supplementary Table S2 - Supplementary Table S2. Diet composition (per 1000 gm).
- Supplementary Figure S7 - Supplementary Figure S7: (A) Food intake of mice on control, 2.5% and 5% St. John's wort extract-supplemented diet at 1 week and 3 week after the introduction of respective diets.
- Supplementary Figure S1 - Supplementary Figure S1: (A) Representative UPLC-ESITOFMS ion chromatogram of the methanolic extract of St. John's wort extract supplemented diet after 20 days of storage at 4C.
- Supplementary Figure S2A - Supplementary Figure S2A: Confirmation of identities of (A) hyperforin and (B) hypericin peaksby comparison of fragmentation pattern with respective authentic standards.
- Supplementary Figure S2B - Supplementary Figure S2B: Confirmation of identities of (A) hyperforin and (B) hypericin peaksby comparison of fragmentation pattern with respective authentic standards.
- Supplementary Figure S3 - Supplementary Figure S3: Putative identification of constituents of St. John's wort extract in the chromatogram of the methanolic extracts by analysis of their fragmentation pattern.
- Supplementary Figure S4 - Supplementary Figure S4: Extracted and integrated ion chromatograms for hyperforin showing the relative abundance (area under the curve of the highlighted peak) do not diminish upon storage of the diet at 4{degree sign}C.
- Supplementary Figure S5 - Supplementary Figure S5: Extracted and integrated ion chromatograms for hypericin showing the relative abundance (area under the curve of the highlighted peak) do not diminish upon storage of the diet at 4{degree sign}C.
- Supplementary Figure S6 - Supplementary Figure S6: Extracted and integrated ion chromatograms for quercetin showing the relative abundance (area under the curve of the highlighted peak) do not diminish upon storage of the diet at 4{degree sign}C.
- Supplementary Figure S8 - Supplementary Figure S8: Serum ALT (A) and ALP (B) level of mice on control, 2.5% and 5% St. John's wort extract supplemented diet. Statistical significance was calculated using oneway ANOVA with Tukey's correction for multiple testing.
- Supplementary Figure S9 - Supplementary Figure S9: Serum ALT (A), ALP (B) and Bile acid (C) levels. All values are presented as mean {plus minus} standard deviation.
- Supplementary Figure S10 - Supplementary Figure S10: Hematoxylin and Eosin-stained sections of colon from saline injected mice on control (Con/Sal) or St. John's wort-supplemented diet (SJW/Sal) and AOM injected mice on control (Con/AOM) or St. John's wort-supplemented diet (SJW/Sal) in the long-term study.
- Supplementary Figure S11 - Supplementary Figure S11: Hematoxylin and Eosin-stained sections of colon from saline injected mice on control (Con/Sal) or St. John's wort-supplemented diet (SJW/Sal) and AOM injected mice on control (Con/AOM) or St. John's wort-supplemented diet (SJW/Sal) in the four week of the short-term study.
Volume 8, Issue 9
