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Abstract: Objective To investigate the role and mechanism of Hexavitamin soya lecithin
on the progression of alcoholic hepatitis. Methods Total of eighteen 4-week-old C57BL/6N mice were divided into 3 groups by the random number table method, with 6 mice in
each group. Mice in alcoholic hepatitis group were intragastrically administered with
30% ethanol at a dose of 5 g/(kg·d), once daily for 8 weeks. Mice in control group were
intragastrically administered with an equal volume of 1 × PBS, once daily for 8 weeks.
Mice in Hexavitamin soya lecithin treatment group were intragastrically administered with
30% ethanol and hexavitin phospholipid [0.8 mg/(g·d)], once daily for 8 weeks. Liver and
jejunum tissues of mice in each group were collected. HE staining and oil red O staining were
used to observe liver tissue pathology. HE staining was used for intestinal tissue pathology.
Immunohistochemical staining was used to analyze the expression level of ZO-1 in intestinal
tissue. Western blot was used to detect the protein expression levels of Occludin and Mucin
2 (MUC2). Intestinal tissue of the mice was performed transcriptomic sequencing, Venn
analysis, KEGG analysis and GSEA analysis. Results Pathological analysis of liver tissue
showed that, compared with the control group, mice in alcoholic hepatitis group exhibited
obvious ballooning degeneration, steatosis, and mild inflammatory cell infiltration in their
liver tissue. Compared with the alcoholic hepatitis group, mice in the Hexavitamin soya
treatment group showed significant improvement in ballooning degeneration and steatosis,
and the inflammatory cell infiltration reduced. Transcriptomic analysis revealed that the
alcoholic hepatitis model group and the six-dimensional phospholipid treatment group shared
34 identical differential genes. KEGG analysis of these differential genes was enriched in
intestinal barrier-related pathways, such as the cell adhesion molecule pathway and the
cytokine-cytokine receptor interaction pathway. These two pathways were significantly
downregulated in alcoholic hepatitis and partially restored after treatment with Hexavitamin
soya lecithin. Immunohistochemical analysis demonstrated that ZO-1 expression of mice reduced
in alcoholic hepatitis group, and after Hexavitamin soya lecithin treatment, the intestinal ZO-1
expression returned to normal levels. HE staining showed that the number of goblet cells in each
intestinal villus of control group, alcoholic hepatitis group and hexa-phosphatidylcholine treatment
group was 11.92 ± 0.88, 8.13 ± 1.30 and 11.40 ± 0.96, respectively. The number of goblet cells
in the small intestine of alcoholic hepatitis group decreased significantly, which was obviously
recovered after Hexavitamin soya lecithin treatment (all P < 0.05). Western blot analysis
showed that the expression of Occludin and MUC2 reduced significantly in the alcoholic
hepatitis group, while the expression levels increased significantly after Hexavitamin soya
lecithin treatment. The differences in relative mRNA expression levels of antibacterial defense
related genes Zbp1 (13.86 ± 5.03 vs. 3.93 ± 1.35 vs. 7.61 ± 2.91), Gbp6 (1.98 ± 0.97 vs. 0.38 ± 0.11 vs.
0.81 ± 0.32), and Irgm2 (22.40 ± 9.02 vs. 5.40 ± 2.05 vs. 10.02 ± 2.25) among the control group,
alcoholic hepatitis group and Hexavitamin soya lecithin treatment group were statistically
significant. Specifically, the expression levels in alcoholic hepatitis group were significantly
lower than those in the control group. After treatment with Hexavitamin soya phospholipids,
the relative mRNA expression levels of Zbp1, Gbp6, and Irgm2 were significantly restored (all
P < 0.05). Conclusion Hexavitamin soya lecithin delay the progression of alcoholic hepatitis
by mitigating alcohol-induced intestinal barrier dysfunction.
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