Fig. 6.
Probiotics VSL#3 pretreatment attenuates morphine tolerance and prevents morphine-induced gut microbiota alterations. (A and B) Morphine analgesic effect was determined after escalating doses of morphine; n = 10 to 20. Ftail flick (3, 51) = 46.78. Fhot plate (3, 51) = 82.50. One-way ANOVA followed by Bonferroni’s correction was used to analyze data. (C) Multidimensional scaling was used to visualize the Bray−Curtis distance of different groups; n = 9 to 15. Data were subjected to PERMANOVA test along with Bonferroni correction. (D) Taxonomic distribution of different groups at phylum level. (E and F) Dot plots show changes in abundance of bacteria in Water+Morphine-treated and VSL#3+Morphine-treated mice at family and genus level using Water+Saline mean proportion as reference; n = 9 to 15. (G) Representative H&E stained intestinal sections from different treatment groups; n = 6 to 10. (H) (i) Representative figure of florescent signal distribution and (ii) summary of serum FITC-dextran concentration from different treatment groups; n = 6 to 10. (I) CFU from liver homogenate of each mouse in different treatment groups; n = 6 to 20. (J−L) Probiotics pretreatment inhibits TLR2 up-regulation by morphine on epithelial and immune cells; n = 11 to 14. (M and N) RT-PCR of the IL-6 gene expression; n = 9 to 19. (E–N) The results were analyzed by one-way ANOVA followed by Bonferroni’s multiple comparisons test. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. Mean ± SD.