Mechanism of Sodium Butyrate Promoting Rumen Epithelial Cell Proliferation in Vitro Based on Insulin-Like Growth Factor Ⅰ Signaling Pathway

ZHANG Yali; LIU Lixiang; SUN Daming; LIU Junhua

doi:10.3969/j.issn.1006-267x.2021.03.049

2021 , Vol. 33 >Issue 3: 1687 - 1698

DOI: https://doi.org/10.3969/j.issn.1006-267x.2021.03.049

Mechanism of Sodium Butyrate Promoting Rumen Epithelial Cell Proliferation in Vitro Based on Insulin-Like Growth Factor Ⅰ Signaling Pathway

ZHANG Yali ,
LIU Lixiang ,
SUN Daming ,
LIU Junhua

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Laboratory of Digestive Tract Microbiology, College of Animal Science and Technology, Nanjing Agricultural University, Jiangsu Key Laboratory of Digestive Tract Nutrition and Animal Health, National Animal Digestive Tract Nutrition International Joint Research Center, Nanjing 210095, China

Received date: 2020-08-19

Online published: 2021-03-18

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Abstract

This experiment was conducted to study the mechanism of sodium butyrate promoting rumen epithelial cell proliferation in vitro based on insulin-like growth factor Ⅰ (IGF-Ⅰ) signaling pathway. Four healthy 49-day-old male lambs with similar parity and body weight were slaughtered to collect rumen epithelial tissue, which was digested with trypsin to obtain primary rumen epithelial cells. After the cells adhered to the wall, they were divided into four groups, and supplemented 0, 2, 4 and 8 mmol/L sodium butyrate, respectively. In order to further study the role of IGF-Ⅰ signaling pathway in the promotion of rumen epithelial cell proliferation by sodium butyrate addition, the cells were divided into three groups, the sodium butyrate group supplemented 4 mmol/L sodium butyrate, the inhibitor group supplemented 4 mmol/L sodium butyrate+2.5 μmol/L insulin-like growth factor Ⅰ receptor (IGF-ⅠR) inhibitor, and the control group supplemented the same volume of DMEM. All cell experiments were repeated 4 times. When the cells were treated for 24 h, the cell fluid was collected for determinate IGF-Ⅰ concentration, and the cells were collected to analyses the cell cycle and expression of genes related to proliferation and apoptosis. The results showed as follows: 1) the proportion of cells in G0/G1 phase in 0 and 8 mmol/L sodium butyrate groups was significantly higher than that in 2 and 4 mmol/L sodium butyrate groups (P<0.05), and the proportion of cells in G0/G1 phase in the 8 mmol/L sodium butyrate group was significantly higher than that in the 0 mmol/L sodium butyrate group (P<0.05). The proportion of cells in S phase in the 4 mmol/L sodium butyrate group was significantly higher than that in 0, 2 and 8 mmol/L sodium butyrate groups (P<0.05). The proportion of cells in G2/M phase in the 8 mmol/L sodium butyrate group was significantly lower than that in 0, 2 and 4 mmol/L sodium butyrate groups (P<0.05). 2) The mRNA relative expression level of cyclin protein A2 (Cyclin A2) in the 8 mmol/L sodium butyrate group was significantly lower than that in 0, 2 and 4 mmol/L sodium butyrate groups (P<0.05), the mRNA relative expression level of cyclin dependent protein kinases Ⅰ (CDKⅠ) in the 8 mmol/L sodium butyrate group was significantly lower than that in the 0 and 2 mmol/L sodium butyrate group (P<0.05), the mRNA relative expression level of cyclin protein DⅠ (Cyclin DⅠ) in 2 and 4 mmol/L sodium butyrate groups was significantly higher than that in 0 and 8 mmol/L sodium butyrate groups (P<0.05), the mRNA relative expression level of cyclin dependent protein kinases 4 (CDK4) in the 4 mmol/L sodium butyrate group was significantly higher than that in the 0, 2 and 8 mmol/L sodium butyrate groups (P<0.05), and the mRNA relative expression level of cyclin dependent protein kinases 6 (CDK6) in 2 and 4 mmol/L sodium butyrate groups was significantly higher than that in 8 mmol/L sodium butyrate group (P<0.05). 3) The mRNA relative expression levels of cysteinyl aspartate specific proteinase-3 (Caspase-3) and B-lymphoma-2-associated X protein (Bax) in the 8 mmol/L sodium butyrate group were significantly higher than those in 0, 2 and 4 mmol/L sodium butyrate groups (P<0.05). 4) The IGF-ⅠR mRNA relative expression level and cell culture fluid IGF-Ⅰ concentration in sodium butyrate group were significantly higher than those in control group and inhibitor group (P<0.05). The mRNA relative expression levels of Cyclin DⅠ and CDK4 in sodium butyrate group were significantly higher than those in control group and inhibitor group (P<0.05). These results suggest that adding 4 mmol/L sodium butyrate promotes the rumen epithelial cell proliferation, mainly through the IGF-Ⅰ signaling pathway, it shows that the mechanism of sodium butyrate promoting rumen epithelial growth is closely related to IGF-Ⅰ signaling pathway.

Key words： sodium butyrate; rumen epithelial cells; proliferation; apoptosis; IGF-Ⅰ; lambs

Cite this article

ZHANG Yali , LIU Lixiang , SUN Daming , LIU Junhua . Mechanism of Sodium Butyrate Promoting Rumen Epithelial Cell Proliferation in Vitro Based on Insulin-Like Growth Factor Ⅰ Signaling Pathway[J]. Chinese Journal of Animal Nutrition, 2021 , 33(3) : 1687 -1698 . DOI: 10.3969/j.issn.1006-267x.2021.03.049

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