Effects of metabolic energy intervention on lipid content and liver transcriptome in finisher yellow-feathered chickens

Figures & data

Table 1. Composition and nutrient levels of experimental diets (% as fed-basis)*.

Components	LME	HME
Ingredient, %
Corn	65.0	80.0
Soybean meal	11.0	11.0
Corn gluten meal	2.70	1.50
Lard	3.00	3.00
L-lysine HCl	0.42	0.40
DL-Methionine	0.22	0.21
L-Threonine	0.13	0.12
Limestone	1.05	1.05
Dicalcium phosphate	1.55	1.55
Salt	0.25	0.25
Premix^a	1.00	1.00
Corncob powder	13.68	0.00
Total	100.00	100.00
Nutrient composition
AME (MJ/kg)	12.06	13.31
CP, %	13.00	13.00
Lys, %	0.83	0.83
Met + Cys, %	0.66	0.66
Ca, %	0.79	0.79
Nonphytate P, %	0.34	0.34

*LME = low metabolic energy; HME = high metabolic energy.

^aPremix provided the following per kilogram of the diet as previously described (Cui et al. 2022).

AME: apparent metabolizable energy; CP: crude protein; Met + Cys: methionine + cysteine; Ca: calcium; P: phosphorus.

Table 2. Effects of low metabolic energy (LME) diet and high metabolic energy (HME) diet on growth performance in Yellow-feathered chickens from 90 to 135 days of age.

Variable*	LME	HME	SEM	P values
Final BW, g	1549^b	1595^a	6.358	0.012
ADG, g	10.16^b	11.56^a	0.121	0.001
ADFI, g	75	75	0	0
Feed/gain, g/g	7.43^a	6.48^b	0.095	0.001

Means within a row with different superscripts (^ab) are different at P < 0.05 (n = 10).

*BW, body weight; ADG, average daily gain; ADFI, average daily feed intake.

Table 3. Effects of low metabolic energy (LME) diet and high metabolic energy (HME) diet on lipid metabolism indices and the relative weight of liver in Yellow-feathered chickens.

Variable*	LME	HME	SEM	P values
Plasma
TG (mmol/L)	1.47^b	2.27^a	0.337	0.032
TCH (mmol/L)	3.12^b	3.67^a	0.264	0.044
HDL-C (mmol/L)	1.81	1.82	0.162	0.953
LDL-C (mmol/L)	0.68	0.76	0.141	0.601
Liver
TG (mmol/g prot)	0.060^b	0.196^a	0.006	0.000
TCH (mmol/g prot)	0.011^b	0.025^a	0.002	0.032
Liver weight (g/100 g of BW)	2.06^b	2.75^a	0.268	0.009

Means within a row with different superscripts (^ab) are different at P < 0.05 (n = 10).

*TG, total triglyceride; TCH, total cholesterol; HDL-C, high density lipoprotein cholesterol; LDL-C, low density lipoprotein cholesterol; BW, body weight.

Figure 1. (a) Volcano map. The red dots represent upregulated (P < 0.05) differentially expressed genes (DEG), the blue dots represent downregulated (P < 0.05) DEG, and the gray dots represent insignificant genes in the liver between chickens fed low metabolic energy (LME) and high metabolic energy (HME) diets. (b) Hierarchical clustering. Cluster analyses were performed based on 480 known DEG in the liver between chickens fed LME and HME diets, the LME diet and HME diet were clustered, respectively. Up-regulated DEG are in red and down-regulated DEG are in green.

Table 4. Lipid metabolism-related biological processes and cellular component identified by Gene Ontology (GO) analysis of differentially expressed genes between chickens fed low metabolic energy (LME) and high metabolic energy (HME) diets.

GO.ID	Term	DEG (HME/LME)	Total	P value	FDR
GO:0034368	Protein-lipid complex remodelling	6	14	9.2E^−07	0.002
GO:0034369	Plasma lipoprotein particle remodelling	6	14	9.2E^−07	0.002
GO:0071827	Plasma lipoprotein particle organisation	6	18	5.2E^−06	0.007
GO:0034374	Low-density lipoprotein particle remodelling	4	6	7.4E^−06	0.007
GO:0071825	Protein-lipid complex subunit organisation	6	20	1E^−05	0.007
GO:0034358	Plasma lipoprotein particle	5	13	2.1E^−05	0.012
GO:1990777	Lipoprotein particle	5	13	2.1E^−05	0.012
GO:0032994	Protein-lipid complex	5	14	3.2E^−05	0.015
GO:0046470	Phosphatidylcholine metabolic process	6	26	5.3E^−05	0.022
GO:0043691	Reverse cholesterol transport	4	9	5.9E^−05	0.022
GO:0055091	Phospholipid homeostasis	4	9	5.9E^−05	0.022
GO:0042632	Cholesterol homeostasis	8	55	1E^−04	0.032
GO:0055092	Sterol homeostasis	8	55	1E^−04	0.032
GO:0097006	Regulation of plasma lipoprotein particle levels	6	31	1.5E^−04	0.040

Figure 2. KEGG pathway analyses of differentially expressed genes (DEG) in the liver between chickens fed LME and HME diets. The x-axis represents the rich factor, the y-axis represents the enriched pathway. The size of the bubble indicates the number of DEG enriched in the pathway, and the different colours represent enrichment significance, the red colour means the enrichment result is significant.

Table 5. The differentially expressed genes involved in five key KEGG pathways between chickens fed low metabolic energy (LME) and high metabolic energy (HME) diets.

Genes	log2FC (HME/LME)	P value
PPAR signalling pathway
FABP3	3.63	0.011
FABP1	4.41	0.004
PLIN1	3.98	0.020
RXRG	1.40	6.15E^−08
Fatty acid elongation
ELOVL2	1.53	1.47E^−09
HADHA	1.35	0.007
ACOT1L	1.04	0.004
Glycerophospholipid metabolism
MBOAT2	6.46	8.71E^−15
PLA2G2E	1.37	0.028
AGPAT2	1.28	0.025
AGPAT3	1.27	0.012
PCYT2	1.13	9.95E^−06
PEMT	1.05	0.001
LCAT	−1.22	0.000
Cytokine-cytokine receptor interaction
PRLR	2.49	0.001
IL-12B	1.44	0.012
CRLF2	1.22	0.010
IL1R2	−2.57	0.006
TNFRSF6B	−2.23	0.048
CCL4	−1.81	0.001
IL21R	−1.61	0.013
CCL1	−1.56	6.02E^−05
IL1R1	−1.53	1.82E^−09
IL12RB2	−1.24	6.57E^−05
CD40LG	−1.19	0.003
CXCL12	−1.13	0.004
AMH	−1.10	0.012
IL18R1	−1.07	0.001
CX3CL1	−1.01	0.009

Figure 3. Validation of RNA-seq data by q-PCR analyses. ** indicate significance at P < 0.01. MBOAT2: membrane bound O-acyltransferase domain containing 2; PLA2G2E: phospholipase A2 group IIE; AGPAT2: 1-acylglycerol-3-phosphate O-acyltransferase 2; AGPAT3: 1-acylglycerol-3-phosphate O-acyltransferase 3; PCYT2: phosphate cytidylyltransferase 2; PEMT: phosphatidylethanolamine N-methyltransferase; ELOVL2: fatty acid elongase 2; HADHA: hydroxyacyl-CoA dehydrogenase trifunctional multienzyme complex subunit alpha; ACOT1L: acyl-CoA thioesterase 1-like; FABP3: fatty acid binding protein 3; FABP1: fatty acid binding protein 1; PLIN1: perilipin 1; RXRG: retinoid X receptor gamma; LCAT: lecithin-cholesterol acyltransferase.

Figure 4. Correlation analysis of the relative expression levels for Log2 Fold Change (FC) of 14 DEG between the RNA-seq and q-PCR.

Cui X, Gou Z, Jiang Z, Li L, Lin X, Fan Q, Wang Y, Jiang S. 2022. Dietary fiber modulates abdominal fat deposition associated with cecal microbiota and metabolites in yellow chickens. Poult Sci. 101(4):101721.

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