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. 2022 Mar;8(2):591-597.
doi: 10.1002/vms3.658. Epub 2021 Oct 20.

In vitro study of chlorine dioxide on porcine intestinal epithelial cell gene markers

Orsolya Palócz  1 , Zoltán Noszticzius  2 , Kristóf Kály-Kullai  2 , Emma Bradley  1 , György Csikó  1
Affiliations

In vitro study of chlorine dioxide on porcine intestinal epithelial cell gene markers

Orsolya Palócz et al. Vet Med Sci. 2022 Mar.

Abstract

Background: Chlorine dioxide (ClO2 ) is an inorganic, potent biocide and is available in highly purified aqueous solution. It can be administered as an oral antiseptic in this form.

Objectives: Our aim is to determine the level of inflammatory markers and cytochrome genes expressed by enterocytes exposed to different concentrations of hyperpure chlorine dioxide solution.

Methods: Porcine jejunal enterocyte cell (IPEC-J2) cultures were treated with the aqueous solution of hyper-pure chlorine dioxide of various concentrations. We determined the alterations in mRNA levels of inflammatory mediators, such as IL6, CXCL8/IL8, TNF, HSPA6 (Hsp70), CAT and PTGS2 (COX2); furthermore, the expression of three cytochrome genes (CYP1A1, CYP1A2, CYP3A29) were analysed by quantitative PCR method.

Results: The highest applied ClO2 concentration reduced the expression of all three investigated CYP genes. The gene expression of PTGS2 and CAT were not altered by most concentrations of ClO2 . The expression of IL8 gene was reduced by all applied concentrations of ClO2 . TNF mRNA level was also decreased by most ClO2 concentrations used.

Conclusions: Different concentrations of chlorine dioxide exhibited immunomodulatory activity and caused altered transcription of CYP450 genes in porcine enterocytes. Further studies are needed to determine the appropriate ClO2 concentration for oral use in animals.

Keywords: chlorine dioxide; cytochrome genes; inflammatory markers; intestinal cells.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

FIGURE 1
FIGURE 1
Viability of porcine jejunal cells (IPEC‐J2) after 15 min of chlorine dioxide (ClO2) treatment. Data expressed as mean ± SD, n = 8/group. The chlorine dioxide was diluted in phosphate‐buffered saline; the 0 mM concentration is the control treatment, contains only phosphate‐buffered saline
FIGURE 2
FIGURE 2
The relative gene expressions of CAT and PTGS2/COX2 at various ClO2 concentrations from 0.07 to 2.22 mM in porcine jejunal cell cultures. Results are expressed as mean mRNA expression ratio relative to controls (n = 6/group). Significant differences are shown in comparison to untreated controls (*p < 0.05, **p < 0.01). Data are shown as means ± SD. CAT, catalase; PTGS2, prostaglandin‐endoperoxide synthase 2, also known as cyclooxygenase 2 (COX2)
FIGURE 3
FIGURE 3
The relative gene expressions of IL6, CXCL8/IL8 and TNF at various ClO2 concentrations from 0.07 to 2.22 mM in porcine jejunal cell cultures. Results are expressed as mean mRNA expression ratio relative to controls (n = 6/group). Significant differences are shown in comparison to untreated controls (*p < 0.05, **p < 0.01). Data are shown as means ± SD. CXCL8, C‐X‐C motif chemokine ligand 8; IL, interleukin; TNF, tumour necrosis factor
FIGURE 4
FIGURE 4
The relative gene expression of HSPA6 at various ClO2 concentrations from 0.07 to 2.22 mM in porcine jejunal cell cultures. Results are expressed as mean mRNA expression ratio relative to controls (n = 6/group). Significant differences are shown in comparison to untreated controls (* and # p < 0.05, **p < 0.01). Data are shown as means ± SD. HSPA6, heat shock protein family A (Hsp70) member 6
FIGURE 5
FIGURE 5
The relative gene expressions of CYP1A1, CYP1A2 and CYP3A29 at various ClO2 concentrations from 0.07 to 2.22 mM in porcine jejunal cell cultures. Results are expressed as mean mRNA expression ratio relative to controls (n = 6/group). Significant differences are shown in comparison to untreated controls (# p < 0.05, ** and ## p < 0.01). Data are shown as means ± SD. CYP, cytochrome P450
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