Effect of vitamin A on barrier function damage of piglet intestinal epithelial cells induced by soybean 7S globulin

doi:10.3969/j.issn.1004-1524.2021.11.04

Acta Agriculturae Zhejiangensis ›› 2021, Vol. 33 ›› Issue (11): 2026-2033.DOI: 10.3969/j.issn.1004-1524.2021.11.04

• Animal Science • Previous Articles Next Articles

Effect of vitamin A on barrier function damage of piglet intestinal epithelial cells induced by soybean 7S globulin

YANG Ju(), DENG Junliang^*(), XIA Jiangying, SONG Tianhao, PANG Lianfeng, REN Zhihua

College of Veterinary Medicine/Key Laboratory of Animal Disease and Human Health of Sichuan Province/Key Laboratory of Environmental Hazards and Animal Diseases of Sichuan Province Colleges and Universities, Sichuan Agriculture University, Chengdu 611130, China

Received:2021-01-08 Online:2021-11-25 Published:2021-11-26
Contact: DENG Junliang

Abstract

Abstract:

To explore effect of vitamin A (V_A) on barrier function damage of piglet intestinal epithelial cells (IPEC-J2) induced by soybean 7S globulin, took IPEC-J2 as the research object and soybean 7S globulin as the inducing factor, cell viability, trans-epithelial electrical resistance (TEER), permeability of sodium fluorescein, cell membrane integrity related indicators and tight junction protein mRNA expression were assessed. The results showed that 5 mg·mL^-1 soybean 7S globulin caused IPEC-J2 viability, TEER, ZO-1, Claudin-1 and Occludin mRNA expression significantly reduced (P<0.01), and permeability of fluorescein sodium, content of alkaline phosphatase (ALP), lactate dehydrogenase (LDH), diamine oxidase (DAO) and intestinal fatty acid binding protein 1 (IFABP1) in the cell culture fluid significantly increased (P<0.01). 0.1 and 1 μmol·L ^-1 V_A could significantly protect cell viability (P<0.01), 0.1-10 μmol·L^-1 V_A could significantly (P<0.05) alleviate the changes of TEER, fluorescein sodium permeability, cell membrane integrity related indicators and tight junction protein mRNA expression caused by 7S globulin; 10 000 μmol·L^-1 V_A exacerbated these effects. The results indicated that 0.1-10 μmol·L^-1 V_A could protect the cell barrier function of IPEC-J2 caused by soybean 7S globulin through protecting cell viability and cell integrity, and affecting expression of tight junction protein mRNA. However, 10 000 μmol·L^-1 V_A aggravated this damage.

Key words: soybean 7S globulin, vitamin A, piglet intestinal epithelial cell, permeability, tight junction protein

CLC Number:

S852.4

YANG Ju, DENG Junliang, XIA Jiangying, SONG Tianhao, PANG Lianfeng, REN Zhihua. Effect of vitamin A on barrier function damage of piglet intestinal epithelial cells induced by soybean 7S globulin[J]. Acta Agriculturae Zhejiangensis, 2021, 33(11): 2026-2033.

Figures/Tables 7

Table 1 Primers for amplification of genes by qRT-PCR

基因名称 Gene name	基因ID Gene ID	上游引物序列(5'-3') Forward primer sequence (5'-3')	下游引物序列(5'-3') Reverse primer sequence (5'-3')	产物大小 Product size/bp
ZO-1	396567	CCAGGGAGAGAAGTGCCAGTAGG	TTTGGTGGGTTTGGTGGGTTGAC	92
Claudin-1	100625166	CCATCGTCAGCACCGCACTG	CGACACGCAGGACATCCACAG	107
Occludin	397236	TGGCTGCCTTCTGCTTCATTGC	GAACACCATCACACCCAGGATAGC	130
β-actin	414396	GGCACCACACCTTCTACAACGAG	TCATCTTCTCACGGTTGGCTTTGG	102

Fig.1 Effect of VA on viability of IPEC-J2 injured by soybean 7S globulin Data on the bars marked without the same uppercase letter indicated significant differences at P<0.01, data on the bars marked without the same lowercase letter indicated significant differences at P<0.05. The same as below.

Fig.2 Trans-epithelial electrical resistance of IPEC-J2

Fig.3 Effect of VA on trans-epithelial electrical resistance of IPEC-J2 monolayer cells induced by soybean 7S globulin

Fig.4 Effect of VA on permeability of sodium fluorescein of IPEC-J2 monolayer cells induced by soybean 7S globulin

Fig.5 Effect of VA on destruction of IPEC-J2 integrity by soybean 7S globulin

Fig.6 Effect of VA on expression of tight junction protein genes in IPEC-J2 induced by soybean 7S globulin

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Effect of vitamin A on barrier function damage of piglet intestinal epithelial cells induced by soybean 7S globulin

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