Probiotic factors partially prevent changes to caspases 3 and 7 activation and transepithelial electrical resistance in a model of 5-fluorouracil-induced epithelial cell damage

Abstract

The potential efficacy of a probiotic-based preventative strategy against intestinal mucositis has yet to be investigated in detail. We evaluated supernatants (SN) from Escherichia coli Nissle 1917 (EcN) and Lactobacillus rhamnosus GG (LGG) for their capacity to prevent 5-fluorouracil (5-FU)-induced damage to intestinal epithelial cells. A 5-day study was performed. IEC-6 cells were treated daily from days 0 to 3, with 1 mL of PBS (untreated control), de Man Rogosa Sharpe (MRS) broth, tryptone soy roth (TSB), LGG SN, or EcN SN. With the exception of the untreated control cells, all groups were treated with 5-FU (5 μM) for 24 h at day 3. Transepithelial electrical resistance (TEER) was determined on days 3, 4, and 5, while activation of caspases 3 and 7 was determined on days 4 and 5 to assess apoptosis. Pretreatment with LGG SN increased TEER (p < 0.05) compared to controls at day 3. 5-FU administration reduced TEER compared to untreated cells on days 4 and 5. Pretreatment with MRS, LGG SN, TSB, and EcN SN partially prevented the decrease in TEER induced by 5-FU on day 4, while EcN SN also improved TEER compared to its TSB vehicle control. These differences were also observed at day 5, along with significant improvements in TEER in cells treated with LGG and EcN SN compared to healthy controls. 5-FU increased caspase activity on days 4 and 5 compared to controls. At day 4, cells pretreated with MRS, TSB, LGG SN, or EcN SN all displayed reduced caspase activity compared to 5-FU controls, while both SN groups had significantly lower caspase activity than their respective vehicle controls. Caspase activity in cells pretreated with MRS, LGG SN, and EcN SN was also reduced at day 5, compared to 5-FU controls. We conclude that pretreatment with selected probiotic SN could prevent or inhibit enterocyte apoptosis and loss of intestinal barrier function induced by 5-FU, potentially forming the basis of a preventative treatment modality for mucositis.

A review of complementary therapies for chemotherapy induced gastrointestinal mucositis

“…Lactobacillus fermentum BR11 reduced jejunal inflammation as evident from reduced myeloperoxidase activity in 5-FU induced rat models of GIM (81). Pre-treatment EcN and LGG (Lactobacillus rhamnosus) supernatants lowered caspase activity, inhibited enterocyte apoptosis and loss of intestinal barrier function induced by 5-FU treatment of IEC-6 cells (82). Oral administration of probiotics Lactobacillus casei variety rhamnosus (Lcr35) or Lactobacillus acidophilus and Bifidobacterium bifidum (LaBi) appeard to ameliorate the intestinal mucositis severity by inhibition the expressions of proinflammatory cytokines (83).…”

Some Contributions to the Study of Oenological Lactic Acid Bacteria through Their Interaction with Polyphenols

“…Bacteria cell-free supernatants of LAB strains have been reported to exhibit functions similar to the living bacteria from which they were derived, and to reduce the infection risk of the use of probiotic bacteria in patients with depressed immune systems [31]. So, CFS from the LAB strains were prepared in MRS broth and their antibacterial activity against E. coli CIAL-153 was evaluated (Figure 2).…”

Newly Developed Synbiotics and the Chemotherapy-Damaged Gut

“…36 Moreover, recently, probiotic-derived factors have begun to show promise at preventing mucositis in vivo and in vitro. 35,37 In addition, there are now early indications that antioxidant and anti-inflammatory constituents in plant extracts, such as Iberogast 38 and grape seed extract, 39 and animal-sourced oils, such as Lyprinol 40 and Emu oil, 41 could have utility in preventing mucositis.…”

“…74 In addition, Prisciandaro et al recently reported that supernatants from Lactobacillus rhamnosus and Escherichia coli Nissle 1917 significantly decreased caspase 3/7 activity after a challenge by the chemotherapy antimetabolite, 5-fluorouracil, in the IEC-6 cell line, showing their potential ability to prevent or inhibit enterocyte apoptosis induced by chemotherapy. 35 …”