Effect of weaning age on intestinal mucosal morphology, permeability, gene expression of tight junction proteins, cytokines and secretory IgA in Wuzhishan mini piglets

Figures & data

Table 1. Sequences of primers used for real-time PCR.

Target	Accession number	Sense and Antisense primer	PCR product (bp)
GAPDH	AF017079	F:5′GAAGGTCGGAGTGAACGGAT3′	149
		R:5′CATGGGTAGAATCATACTGGAACA3′
Occludin	NM_001163647.1	F:5′ATGCTTTCTCAGCCAGCGTA3′	176
		R:5′AAGGTTCCATAGCCTCGGTC3′
ZO-1	XM003353439.1	F:5′GAGGATGGTCACACCGTGGT3′	169
		R:5′GGAGGATGCTGTTGTCTCGG3′
Claudin-1	NM_001161635.1	F:5′GGACTAATAGCCATCTTTGT3′	88
		R:5′CAGCCATCCGCATCTTCT3′
IL-1β	NM_214055.1	F:5′ACCTGGACCTTGGTTCTC3′	124
		R:5′GGATTCTTCATCGGCTTC3′
TNF-α	NM_214022.1	F:5′ACGCTCTTCTGCCTACTGC3′	162
		R:5′TCCCTCGGCTTTGACATT3′
IL-10	NM_214041.1	F:5′CACTGCTCTATTGCCTGATCTTCC3′	136
		R:5′AAACTCTTCACTGGGCCGAAG3′
IL-6	NM_214399.1	F:5’TTCAGTCCAGTCGCCTTCT3’	91
		R:5’GTGGCATCACCTTTGGCATCTTCTT3’

GAPDH: glyceraldehyde phosphate dehydrogenase; ZO-1: zonula occluden-1; IL-1β: interleukin 1β; TNF-α: tumor necrosis factor α; IL-10: interleukin 10; IL-6: interleukin 6.

Table 2. Effect of weaning age on jejunal and ileal morphology of Wuzhishan piglets at 56 days old.

Items	Treatments^a				SEM^b	p
	21W	28W	35W	42W
Jejunum
Villus height (μm)	239.89	245.33	258.24	274.35	8.202	.512
Villus width (μm)	127.64	114.56	108.58	102.25	4.401	.212
Crypt depth (μm)	137.51	135.17	126.44	130.46	5.277	.914
Villus hight:crypt depth (V/C)	1.75	1.83	2.07	2.11	0.065	.122
Ileum
Villus height (μm)	214.32	208.02	235.96	242.92	8.396	.443
Villus width (μm)	115.58	112.68	99.25	98.71	4.511	.462
Crypt depth (μm)	132.71	131.33	129.06	128.67	5.988	.996
Villus hight:crypt depth (V/C)	1.64	1.60	1.85	1.90	0.069	.333

^a Piglets were weaned at 21(21W), 28(28W), 35(35W) and 42(42W) days old, respectively.

^b SEM, total standard error of means (n = 8).

Figure 1. Effects of weaning age (21W, 28W, 35W, 42W) on plasma D-lactate level (A) and DAO activity (B) in piglets at 28, 35, 42, 49 and 56 day old (n = 8 pigs per treatment). ^a,b,cDifferent letters within a row indicate a significant difference (p < .05) between them. Values are means, with standard deviation represented by vertical bars.

Table 3. Effect of weaning age on the relative expression levels of Claudin-1, Occludin and ZO-1 mRNA in the intestine mucosa of Wuzhishan piglets at 56 days old.

items	Treatments^d				SEM^e	p
	21W	28W	35W	42W
Jejunum
Occludin	1.00^b	1.32^a,b	1.55^a	1.67^a	0.095	.005
ZO-1	1.00^c	1.54^b	1.89^a,b	2.01^a	0.130	.001
Claudin	1.00	1.21	1.23	1.30	0.079	.254
Ileum
Occludin	1.00^b	1.48^a,b	1.71^a	1.95^a	0.119	.004
ZO-1	1.00^b	1.21^b	1.86^a	2.23^a	0.160	.001
Claudin	1.00^b	1.23^a,b	1.44^a	1.59^a	0.087	.009

^a,b,c Different letters within a row indicate a significant difference (p < .05) between them.

^d Piglets were weaned at 21(21W), 28(28W), 35(35W) and 42(42W) days old, respectively.

^e SEM, total standard error of means (n = 8).

Table 4. Effects of weaning age on the relative expression levels of cytokines mRNA in intestinal mucosa of Wuzhishan piglets at 56 days old.

Items	Treatments^a				SEM^b	p
	21W	28W	35W	42W
Jejunum
IL-1β	1.00	0.88	0.80	0.79	0.064	.359
TNF-α	1.00	0.96	0.80	0.75	0.075	.363
IL-6	1.00	0.84	0.74	0.78	0.046	.180
IL-10	1.00	1.04	1.25	1.32	0.125	.142
Ileum
IL-1β	1.00	0.95	0.82	0.79	0.087	.292
TNF-α	1.00	0.84	0.80	0.80	0.092	.612
IL-6	1.00	0.86	0.75	0.78	0.121	.315
IL-10	1.00	1.34	1.43	1.47	0.158	.130

IL-1β: interleukin 1β; TNF-α: tumor necrosis factor α; IL-10: interleukin 10; IL-6: interleukin 6.

^a Piglets were weaned at 21(21W), 28(28W), 35(35W) and 42(42W) days old, respectively.

^b SEM, total standard error of means (n = 8).

Table 5. Effects of weaning age on contents of intestinal mucus sIgA in Wuzhishan piglets at 56 days old (μg/g).

items	Treatments^d				SEM^e	p
	21W	28W	35W	42W
Jejunum	67.34^a	61.01^a,b	52.25^b,c	47.31^c	2.677	.007
Ileum	72.84^a	71.08^a	57.50^b	54.51^b	2.819	.011

^a,b,c Different letters within a row indicate a significant difference (p < .05) between them.

^d Piglets were weaned at 21(21W), 28(28W), 35(35W) and 42 (W) days old, respectively.

^e SEM, total standard error of means (n = 8).