Protective effects of curcumin on laying hens fed soybean meal with heat-induced protein oxidation

Figures & data

Table 1. Composition and nutrient level of the basal diet (g/kg, as fed basis unless otherwise stated).

Items	Contents
	CON	HSBM
Ingredients
Maize	620	620
Soybean meal	240	—
Heated soybean meal	—	240
Soybean oil	10.0	10.0
Limestone	80.0	80.0
Premix^a	50.0	50.0
Calculated nutrient levels
Apparent metabolisable energy (MJ/kg)	11.2	11.2
Crude protein	160	160
Calcium	35.8	35.8
Total phosphorus	6.17	6.17
Available phosphorus	3.69	3.69
Lysine	7.92	7.92
Methionine	3.53	3.53
Total sulphur amino acids	6.33	6.33

CON: laying hens fed a basal diet; HSBM: laying hens fed a basal diet containing heated soybean meal.

^aPremix provided per kilogram of diet: transretinyl acetate, 10,000 U; cholecalciferol, 3000 U; all-rac-α-tocopherol, 30.0 U; menadione, 1.00 mg; thiamine, 1.00 mg; riboflavin, 6.00 mg; nicotinamide, 40.0 mg; choline chloride, 350 mg; calcium pantothenate, 10.0 mg; pyidoxine HCl, 3.00 mg; biotin, 0.100 mg; folic acid, 0.300 mg; cobalamine, 0.010 mg; Cu (copper sulfate), 8.00 mg; Fe (ferrous sulfate), 80.0 mg; Mn (manganese sulfate), 100 mg; I (calcium iodate), 1.00 mg; Se (sodium selenite), 0.300 mg; calcium, 6.25 g; phosphorus, 3.00 g; methionine, 1.00 g.

Table 2. Sequences for real-time PCR primers.

Items	Gene bank ID	Primer sequence, sense/antisense
ZO1	XM_015278975.2	TGTAGCCACAGCAAGAGGTG
		CTGGAATGGCTCCTTGTGGT
OCLN	NM_205128.1	CCGTAACCCCGAGTTGGAT
		ATTGAGGCGGTCGTTGATG
CLDN1	NM_001013611.2	GCAGATCCAGTGCAAGGTGTA
		CACTTCATGCCCGTCACAG
CLDN2	NM_001277622.1	CCTGCTCACCCTCATTGGAG
		GCTGAACTCACTCTTGGGCT
CLDN3	NM_204202.1	CCCGTCCCGTTGTTGTTTTG
		CCCCTTCAACCTTCCCGAAA
B^0AT	XM_419056.6	GGGTTTTGTGTTGGCTTAGGAA
		TCCATGGCTCTGGCAGAGATTT
LAT1	NM_001030579.2	CACACTATGGGCGCATGCT
		ATTGTGCCTGGAGGTGTTGGT
ASCT1	XM_001232899.5	TTGGCCGGGAAGGAGAAG
		AGACCATAGTTGCCTCATTGAATG
EAAT3	XM_424930.6	TGCTGCTTTGGATTCCAGTGT
		AGCAATGACTGTAGTGCAGAAGTAATATATG
b^0,+AT	NM_001199133.1	CAGTAGTGAATTCTCTGAGTGTGAAGCT
		GCAATGATTGCCACAACTACCA
y^+LAT2	XM_015292306.2	GCCCTGTCAGTAAATCAGACAAGA
		TTCAGTTGCATTGTGTTTTGGTT
CAT1	NM_001145490.1	CAAGAGGAAAACTCCAGTAATTGCA
		AAGTCGAAGAGGAAGGCCATAA
CAT2	NM_001199102.1	TGCTCGCGTTCCCAAGA
		GGCCCACAGTTCACCAACAG
PepT1	NM_204365.1	CCCCTGAGGAGGATCACTGTT
		CAAAAGAGCAGCAGCAACGA
GLUT2	NM_207178.1	CTCAGCCAGGTGTACTGTGCTT
		CGTCATCCGCTTCAGTCTCA
SGLT1	NM_001293240.1	GCCATGGCCAGGGCTTA
		CAATAACCTGATCTGTGCACCAGTA
β-actin	NM_205518.1	TGCTGTGTTCCCATCTATCG
		TTGGTGACAATACCGTGTTCA

ZO1: zonula occludens 1; OCLN: occludin; CLDN1: claudin 1; CLDN2: claudin 2; CLDN3: claudin 3; B⁰AT: B⁰ amino acid transporter; LAT1: L-type amino acid transporter 1; ASCT1: alanine-serine-cysteine-threonine transporter 1; EAAT3: excitatory amino acid transporter 3; b^0,+AT: b^0,+ amino acid transporter; y⁺LAT2: y⁺L amino acid transporter 2; CAT1: cationic amino acid transporter 1; CAT2: cationic amino acid transporter 2; PepT1: oligopeptide transporter 1; GLUT2: glucose transporter 2; SGLT1: sodium-dependent glucose transporter 1.

Table 3. Effects of dietary curcumin supplementation on serum diamine oxidase activity and D-lactate concentration of laying hens fed heated soybean meal.

Items	CON	HSBM	HSBM + Cur	SEM^1	p-Value
DAO (U/L)	8.51^b	13.2^a	13.0^a	0.604	<.001
D-LA (mmol/L)	0.714^b	1.28^a	0.882^b	0.066	<.001

CON: laying hens fed a soybean meal-corn basal diet; HSBM: laying hens fed a heated soybean meal-corn basal diet; HSBM + Cur: laying hens fed a heated soybean meal-corn basal diet supplemented with 150 mg/kg curcumin; DAO: diamine oxidase; D-LA: D-lactate.

¹SEM: standard errors of mean.

^a,bMean values within a row with different superscripts letters are significantly different at p < .05.

Table 4. Effects of dietary curcumin supplementation on mRNA expression of genes related to intestinal barrier of laying hens fed heated soybean meal.

Items	CON	HSBM	HSBM + Cur	SEM^1	p-Value
Jejunum
ZO1	1.00	0.962	1.05	0.064	.873
OCLN	1.00^a	0.632^b	0.651^b	0.061	.016
CLDN1	1.00	0.969	0.939	0.075	.948
CLDN2	1.00	0.909	0.859	0.071	.738
CLDN3	1.00^a	0.674^b	1.01^a	0.053	.007
Ileum
ZO1	1.00	0.944	1.08	0.067	.725
OCLN	1.00^a,b	0.707^b	1.10^a	0.065	.026
CLDN1	1.00	1.05	0.956	0.093	.930
CLDN2	1.00	0.847	0.914	0.064	.645
CLDN3	1.00^a	0.648^b	0.864^a,b	0.056	.025

CON: laying hens fed a soybean meal-corn basal diet; HSBM: laying hens fed a heated soybean meal-corn basal diet; HSBM + Cur: laying hens fed a heated soybean meal-corn basal diet supplemented with 150 mg/kg curcumin; ZO1: zonula occludens 1; OCLN: occludin; CLDN1: claudin 1; CLDN2: claudin 2; CLDN3: claudin 3.

¹SEM: standard errors of mean.

^a,bMean values within a row with different superscripts letters are significantly different at p < .05.

Table 5. Effects of dietary curcumin supplementation on serum biochemical parameters of laying hens fed heated soybean meal.

Items	CON	HSBM	HSBM + Cur	SEM^1	p-Value
Glucose (mmol/L)	15.9^a	16.3^a	13.2^b	0.422	.001
Total protein (g/L)	55.0^a	47.7^b	51.8^a,b	1.57	.013
Albumin (g/L)	21.2^a	18.0^b	20.1^a	0.549	.002
Uric acid (µmol/L)	196	237	201	8.22	.077

CON: laying hens fed a soybean meal-corn basal diet; HSBM: laying hens fed a heated soybean meal-corn basal diet; HSBM + Cur: laying hens fed a heated soybean meal-corn basal diet supplemented with 150 mg/kg curcumin.

¹SEM: standard errors of mean.

^a,bMean values within a row with different superscripts letters are significantly different at p < .05.

Table 6. Effects of dietary curcumin supplementation on contents of amino acids in the serum of laying hens fed heated soybean meal (mg/L).

Items	CON	HSBM	HSBM + Cur	SEM^1	p-Value
Aspartic acid	3.29	4.01	5.17	0.380	.140
Threonine	26.2	38.1	32.9	2.24	.081
Serine	46.8	44.8	55.4	2.12	.111
Glutamic acid	26.9	29.2	28.3	1.50	.837
Glycine	23.5	24.6	26.9	0.840	.283
Alanine	28.3	30.5	35.1	1.29	.124
Valine	15.4^b	21.5^a	13.9^b	1.14	.003
Methionine	9.62^b	12.0^a	9.68^b	0.488	.047
Isoleucine	8.69^b	11.3^a	7.77^b	0.505	.003
Leucine	20.5^b	23.9^a	18.6^b	0.729	.002
Tyrosine	18.5	25.4	21.1	1.62	.214
Phenylalanine	21.4	27.3	26.2	1.21	.109
Lysine	33.8	48.6	42.5	2.74	.072
Histidine	12.2^b	14.3^a,b	15.4^a	0.475	.012
Arginine	47.9	54.2	51.8	2.28	.559
Proline	17.5	19.1	16.6	0.651	.287

CON: laying hens fed a soybean meal-corn basal diet; HSBM, laying hens fed a heated soybean meal-corn basal diet; HSBM + Cur: laying hens fed a heated soybean meal-corn basal diet supplemented with 150 mg/kg curcumin.

¹SEM: standard errors of mean.

^a,bMean values within a row with different superscripts letters are significantly different at p < .05.

Table 7. Effects of dietary curcumin supplementation on contents of amino acids in the liver of laying hens fed heated soybean meal (mg/g).

Items	CON	HSBM	HSBM + Cur	SEM^1	p-Value
Aspartic acid	14.9	14.0	14.6	0.315	.157
Threonine	7.49	7.02	7.45	0.184	.195
Serine	7.91	7.44	8.49	0.335	.258
Glutamic acid	20.7^a	19.3^b	20.3^a,b	0.337	.029
Glycine	7.72^a	7.20^b	7.26^b	0.144	.036
Alanine	9.10^a	8.53^b	8.82^a,b	0.140	.034
Valine	8.22	7.75	8.13	0.185	.207
Methionine	3.37	3.07	3.34	0.126	.211
Isoleucine	6.26	5.85	6.28	0.171	.191
Leucine	13.5	12.7	13.3	0.312	.170
Tyrosine	5.78^a	5.28^b	5.76^a	0.133	.023
Phenylalanine	7.21	6.83	7.13	0.154	.208
Lysine	12.1	11.4	11.9	0.258	.177
Histidine	3.94	3.71	3.81	0.085	.173
Arginine	9.96^a	9.22^b	9.84^a	0.192	.042
Proline	6.55	6.14	6.39	0.135	.126

CON, laying hens fed a soybean meal-corn basal diet; HSBM, laying hens fed a heated soybean meal-corn basal diet; HSBM + Cur, laying hens fed a heated soybean meal-corn basal diet supplemented with 150 mg/kg curcumin.

¹SEM: standard errors of mean.

^a,bMean values within a row with different superscripts letters are significantly different at p < .05.

Table 8. Effects of dietary curcumin supplementation on contents of amino acids in the egg yolk of laying hens fed heated soybean meal (mg/g).

Items	CON	HSBM	HSBM + Cur	SEM^1	p-Value
Aspartic acid	14.1	13.0	13.6	0.401	.204
Threonine	7.69	7.04	7.43	0.240	.201
Serine	12.6^a	10.9^b	12.1^a	0.368	.010
Glutamic acid	18.5	17.2	17.6	0.517	.231
Glycine	4.65	4.68	4.71	0.223	.981
Alanine	7.78	7.26	7.44	0.233	.330
Valine	8.06	7.40	7.79	0.238	.203
Methionine	3.38^a	3.01^b	3.53^a	0.108	.009
Isoleucine	6.82^a	6.09^b	6.54^a,b	0.185	.037
Leucine	12.9	11.9	12.4	0.373	.227
Tyrosine	6.31^a	5.67^b	6.17^a	0.124	.005
Phenylalanine	6.37	5.92	6.23	0.174	.248
Lysine	11.3	10.4	10.8	0.336	.218
Histidine	3.51	3.25	3.31	0.121	.332
Arginine	10.1^a	9.08^b	9.58^a,b	0.247	.035
Proline	6.17^a	5.49^b	5.96^a,b	0.154	.029

CON: laying hens fed a soybean meal-corn basal diet; HSBM: laying hens fed a heated soybean meal-corn basal diet; HSBM + Cur: laying hens fed a heated soybean meal-corn basal diet supplemented with 150 mg/kg curcumin.

¹SEM: standard errors of mean.

^a,bMean values within a row with different superscripts letters are significantly different at p < .05.

Table 9. Effects of dietary curcumin supplementation on mRNA expression of nutrient transporters in the intestinal mucosa of laying hens fed heated soybean meal.

Items	CON	HSBM	HSBM + Cur	SEM^1	p-Value
Jejunum
B^0AT	1.00	0.972	1.04	0.077	.942
LAT1	1.00	1.04	1.06	0.097	.968
ASCT1	1.00	0.992	1.01	0.086	.996
EAAT3	1.00^a	0.732^b	0.734^b	0.045	.011
b^0,+AT	1.00	0.942	0.976	0.070	.951
y^+LAT2	1.00^a,b	0.638^b	1.13^a	0.079	.023
CAT1	1.00^a	0.736^b	1.06^a	0.044	.002
CAT2	1.00	0.902	0.972	0.085	.897
PepT1	1.00^a	0.483^c	0.739^b	0.057	<.001
GLUT2	1.00^a	0.730^b	0.680^b	0.049	.008
SGLT1	1.00	0.853	1.25	0.073	.078
Ileum
B^0AT	1.00^a	0.569^b	1.04^a	0.051	<.001
LAT1	1.00^a	0.626^b	0.946^a	0.049	.001
ASCT1	1.00	0.832	0.927	0.098	.796
EAAT3	1.00	0.909	0.888	0.077	.832
b^0,+AT	1.00	0.887	0.920	0.078	.843
y^+LAT2	1.00^a	0.685^b	0.993^a	0.051	.009
CAT1	1.00^b	0.879^b	1.23^a	0.043	.002
CAT2	1.00	0.991	0.912	0.056	.796
PepT1	1.00	0.908	0.982	0.074	.879
GLUT2	1.00	0.882	0.848	0.083	.747
SGLT1	1.00^a	0.985^a	0.673^b	0.051	.008

CON: laying hens fed a soybean meal-corn basal diet; HSBM: laying hens fed a heated soybean meal-corn basal diet; HSBM + Cur: laying hens fed a heated soybean meal-corn basal diet supplemented with 150 mg/kg curcumin; B⁰AT: B⁰ amino acid transporter; LAT1: L-type amino acid transporter 1; ASCT1: alanine-serine-cysteine-threonine transporter 1; EAAT3: excitatory amino acid transporter 3; b^0,+AT: b^0,+ amino acid transporter; y⁺LAT2: y⁺L amino acid transporter 2; CAT1: cationic amino acid transporter 1; CAT2: cationic amino acid transporter 2; PepT1: oligopeptide transporter 1; GLUT2: glucose transporter 2; SGLT1: sodium-dependent glucose transporter 1.

¹SEM: standard errors of mean.

^a,b,cMean values within a row with different superscripts letters are significantly different at p < .05.