The protective effect of VSL#3 on intestinal permeability in a rat model of alcoholic intestinal injury

Bing Chang, Lixuan Sang, Ying Wang, Jing Tong, Dai Zhang, Bingyuan Wang, Bing Chang, Lixuan Sang, Ying Wang, Jing Tong, Dai Zhang, Bingyuan Wang

Abstract

Background: This study aimed to investigate the mechanism of the probiotic VSL#3 in acute alcoholic intestinal injury, and evaluate the effect of VSL#3, glutamine,VSL#3+glutamine and heat-killed VSL#3 therapy in a rat model.

Methods: Six- to eight-week-old male wild-type rats were divided into seven groups. To establish the acute alcohol liver disease model, rats received three doses of corn starch dissolved in PBS/40% alcohol administered intra-gastrically every 12 hours. Treatment groups received an intra-gastric dose of VSL#3, Glutamine, heat-killed VSL#3, or VSL#3+Glutamine 30 minutes prior to alcohol administration. The placebo group was treated with PBS prior to alcohol administration. TNFα and endotoxin in plasma was measured by ELISA and Tachypleus Ameboctye Lysate assays, and electron microscopy, Western blotting, and reverse transcription polymerase chain reaction were used to identify the mechanisms of VSL#3 in the regulation of epithelial permeability.

Results: First, compared with control group, endotoxin and TNFα in alcohol group was obviously high. At the same time, in VSL#3 group,the expression of endotoxin and TNFα obviously lower than the alcohol group. And the trends of the expression of tight junction proteins in these groups were reversed with the change of endotoxin and TNFα. Second, compared the groups of VSL#3 with glutamine,VSL#3+glutamine and heat-killed VSL#3,we found that both VSL#3 and heat-killed VSL#3, glutamine were as effective as VSL#3+glutamine in the treatment of acute alcohol liver disease, the expression of endotoxin and TNFα were lower than the alcohol group, and tight junction proteins were higher than the alcohol group whereas the expression of tight junction proteins were higher in VSL#3 + glutamine group than either agent alone, but have no significant difference.

Conclusion: We conclude that VSL#3 treatment can regulate the ecological balance of the gut microflora, preventing passage of endotoxin and other bacterial products from the gut lumen into the portal circulation and down-regulating the expression of TNFα, which could otherwise down-regulate the expression of tight junction proteins and increase epithelial permeability.

Figures

Figure 1
Figure 1
The expression of tight junctions and microvilli of small intestine cell under electron microscopy. (A) The expression of tight junctions and microvili in control group; (B) The expression of tight junctions and microvili in alcohol group; (C) The expression of tight junctions and microvili in glutamine group; (D) The expression of tight junctions and microvili in VSL#3 group; (E) The expression of tight junctions and microvili in VSL#3+glutamine group; (F) The expression of tight junctions and microvili in placebo group.
Figure 2
Figure 2
The expression of TNFα in each group measured by ELISA. *P < 0.05 versus alcohol group.
Figure 3
Figure 3
Levels of endotoxin in each group measured using the Tachypleus Amebocye Lysate assay. *P < 0.05 versus alcohol group.
Figure 4
Figure 4
Western blots of tight junction proteins (occludin and ZO-1). 1. control group; 2. alcohol group; 3. Glutamine group; 4. VSL#3 group; 5. VSL#3 + Glutamine group; 6. heat-killed VSL#3 group; 7. placebo group.
Figure 5
Figure 5
Western blot densitometry of tight junction protein (occludin and ZO-1) levels in each group. (A) Western blot densitometry of occludin; (B) Western blot densitometry of ZO-1.
Figure 6
Figure 6
RT-PCR to measure expression of mRNAs encoding tight junction proteins (occludin and ZO-1). 1. control group; 2.alcohol group; 3. Glutamine group; 4. VSL#3 group; 5. VSL#3 + Glutaminegroup; 6. heat-killed VSL#3 group ; 7. placebo group.
Figure 7
Figure 7
PCR densitometry of mRNA encoding tight junction proteins (occludin and ZO-1) in each group. (A) RT-PCR densitometry of occludin mRNA encoding; (B) RT-PCR densitometry of ZO-1 mRNA encoding.

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