Probiotic OMNi-BiOTiC® 10 AAD Reduces Cyclophosphamide-Induced Inflammation and Adipose Tissue Wasting in Mice

Abstract

Cancer therapy is often associated with severe side effects such as drug induced weight loss, also known as chemotherapy-induced cachexia. The aim of this study was to investigate the effects of a multispecies probiotic (OMNi-BiOTiC^® 10 AAD) in a chemotherapy mouse model. A total of 24 male BALB/c mice were gavage-fed with the probiotic formulation or water, once a day for 3 weeks. In the third week, the mice received intraperitoneal cyclophosphamide. At euthanasia, the organs were dissected, and serum was sampled for cytokine analysis. Tight junction components, myosin light chain kinase, mucins, and apoptosis markers were detected in the ileum and colon using histological analyses and qRT-PCR. Lipolysis was analyzed by enzymatic activity assay, Western blotting analyses, and qRT-PCR in WAT. The fecal microbiome was measured with 16S-rRNA gene sequencing from stool samples, and fecal volatile organic compounds analysis was performed using gas chromatography/mass spectrometry. The probiotic-fed mice exhibited significantly less body weight loss and adipose tissue wasting associated with a reduced CGI58 mediated lipolysis. They showed significantly fewer pro-inflammatory cytokines and lower gut permeability compared to animals fed without the probiotic. The colons of the probiotic-fed animals showed lower inflammation scores and less goblet cell loss. qRT-PCR revealed no differences in regards to tight junction components, mucins, or apoptosis markers. No differences in microbiome alpha diversity, but differences in beta diversity, were observed between the treatment groups. Taxonomic analysis showed that the probiotic group had a lower relative abundance of Odoribacter and Ruminococcus-UCG014 and a higher abundance of Desulfovibrio. VOC analysis yielded no significant differences. The results of this study indicate that oral administration of the multispecies probiotic OMNi-BiOTiC^® 10 AAD could mitigate cyclophosphamide-induced chemotherapy side effects.

Keywords: chemotherapy; gut permeability; histomorphometry; inflammation; microbiome; volatile organic compound; weight loss.

Figure 1

Nine-week-old mice received a daily gavage of either 10 AAD (AAD) or water (Aqua) for 4 weeks. (a) At delivery, mice were weighed (body weight delivery) and treated with CTX for one week. (b) Body weights at euthanasia. (c) Daily calorie intake during CTX therapy was determined by weighing the amount of food in the hoppers to estimate the mean daily calorie intake of the two groups. Data are shown as median ± interquartile range (n = 9 per group).

Figure 2

Tissue weights of total (a), perirenal (b), gonadal (c), and inguinal (d) WAT of 10 AAD or water (Aqua) treated animals after 1 week of CTX treatment. Western blotting analysis of ATGL (e); HSL, pHSL (Ser563) (f); and CGI-58 (g) in WAT Triglyceride hydrolase (TGH); activity of WAT (h); Western blot (i). Data are shown as median ± interquartile range (n = 9 per group (a–d); n = 6 per group (e–i)).

Figure 3

After 3 weeks of 10 AAD/Aqua treatment, the mice were treated with CTX for another week. Thereafter, the mice were sacrificed. FITC permeability (a); histological slides of ileum samples of the Aqua (b) and 10 AAD (c) group (H&E staining, 400× magnification). Histomorphometry markers for inflammation (d,e,g,h) in the colon. Correlation analysis (f) (only significant correlations are displayed). Correlation paths (i) and colon histology (H&E staining, 400× magnification) of Aqua (j) and 10 AAD (k). Data are shown as median ± interquartile range (n = 9 per group).

Figure 4

Microbiome analysis. Markers for alpha (a–c) and beta (d,e) diversity. Mean relative abundances at the phylum (f) and family (g) level. Distribution of the mean relative abundances between the groups at the phylum (h) and family (i) level. * p < 0.05.

Figure 5

Charts (median and interquartile range) of the relative abundance of species with significant group differences (a–c); correlation analysis (d) (only significant correlations displayed), and correlation path analysis (e).