丁酸钠和吲哚-3-丙酸共处理对Caco-2细胞紧密连接以及炎症因子的影响

doi:10.11843/j.issn.0366-6964.2024.11.028

Abstract

Abstract:

This study aims to investigate the effects of co-treatment of sodium butyrate and indole-3-propionic acid on the expression of tight junction-related proteins and inflammatory factors in Caco-2 cells. Caco-2 cells were administrated with 0.4 mmol·L^-1 sodium butyrate and 0.1 mmol·L^-1 indole-3-propionic acid each and together for 24 h; Then the transmembrane resistance (TEER) value, expression of tight junction related proteins, gene expression and secretion level of inflammatory cytokines were measured. The results showed that, compared with control group, the co-treatment of sodium butyrate and indole-3-propionic acid significantly increased the TEER value in Caco-2 cells (P < 0.01), while no significant changes were observed in sodium butyrate or indole-3-propionic group (P>0.05); In addition, the results by qPCR and Western blot showed that the relative expression of Claudin-2 mRNA was significantly down-regulated in the co-treatment group compared with the control group (P < 0.01); Compared with sodium butyrate group, the relative mRNA expression of Claudin-1 was significantly up-regulated in the co-treatment group (P < 0.05), and the protein expression of Claudin-1, ZO-1 and Occludin was highly significantly increased in the co-treatment group(P < 0.001); Compared with the indole-3-propionic acid group, the relative mRNA expression of ZO-1 was significantly up-regulated in the co-treatment group (P < 0.05), and the protein expression of Claudin-1 and ZO-1 was highly significantly increased (P < 0.001). For inflammatory cytokines, the relative mRNA expression levels of IL-6, IL-1β, IL-8 and TNF-α were significantly down-regulated in the co-treatment group (P < 0.05); Meanwhile, the results of ELISA showed that the co-treatment group highly significantly suppressed the secretion of IL-6, IL-1β, IL-8 and TNF-α in the cell supernatant compared with the control group (P < 0.001). In conclusion, compared with sodium butyrate or indole-3-propionic acid, the co-treatment of sodium butyrate and indole-3-propionic acid increased the TEER value, promoted the expression of tight junction proteins, suppressed the secretion of pro-inflammatory cytokines, and reduced intestinal permeability. Overall, these results suggested that the co-treatment of sodium butyrate and indole-3-propionic acid has a stronger role in enhancing intestinal epithelial cell barrier function and inhibiting inflammatory response.

Key words: sodium butyrate, indole-3-propionic acid, tight junction, inflammatory cytokines, Caco-2 cells

CLC Number:

S816

Kexin WANG, Xian WU, Haizhou GONG, Qiaoyu FANG, Xiangchen LI, Yanan ZHANG. Effects of Co-Treatment of Sodium Butyrate and Indole-3-Propionic Acid on Tight Junctions and Inflammatory Cytokines in Caco-2 Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(11): 5124-5134.

Figures/Tables 7

Table 1

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References 33

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基因名称 Gene name	引物序列(5'→3') Primer sequence(5'→3')	参考文献 Reference
Occludin	F: GGGCATTGCTCATCCTGAAG R: GCCTGTAAGGAGGTGGACTT	[12]
ZO-1	F: TTCACGCAGTTACGAGCAAG R: TTGGTGTTTGAAGGCAGAGC	[12]
Claudin-1	F: CCAGGTACGAATTTGGTCAGG R: TGGTGTTGGGTAAGAGGTTGT	[14]
Claudin-2	F: CCTAAACCACAAGGCTAAGGGCTA R: GGTGTCTTCCTCAGTACCCCACA	[15]
IL-6	F: TGGCAGCCTTCCTGATTTCT R: AATTTCTGTGTTGGCGCAGT	[12]
IL-1β	F: GAATGACGCCCTCAATCAAAGT R: TCATCTTGGGCAGTCACATACA	[14]
IL-8	F: CCTGAACCTTCCAAAGATGGC R: TTCACCAGGCAAGTCTCCTCA	[14]
TNF-α	F: CTGCCTGCTGCACTTTGGAG R: ACATGGGCTACAGGCTTGTCACT	[12]
β-Actin	F: TGCAGAAAGAGATCACCGC R: CCGATCCACACCGAGTATTTG	[16]

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Effects of Co-Treatment of Sodium Butyrate and Indole-3-Propionic Acid on Tight Junctions and Inflammatory Cytokines in Caco-2 Cells

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