基于转录组测序研究高精料饲粮中添加苦豆子总碱对绵羊肝脏糖代谢的影响

doi:10.16431/j.cnki.1671-7236.2026.02.015

Abstract

Abstract:

Objective To investigate the mechanism by which liver injury in sheep was alleviated and the key pathways of glucose metabolism were regulated through the addition of total alkaloids from Sophora alopecuroides to high-concentrate diets based on transcriptome sequencing. Method Eighteen 3-month-old Dumont male sheep were randomly divided into three groups： G1 （concentrate to roughage ratio was 50∶50）， G2 （concentrate to roughage ratio was 70∶30）， and S3 （concentrate to roughage ratio was 70∶30， added with 121 mg/kg total alkaloids of Sophora alopecuroides）. After a 15-day pre-feeding period and a 60-day experimental period， three sheep were randomly selected from each group for slaughter. Liver tissue was collected for transcriptomic sequencing， with differentially expressed genes （DEGs） identified and subjected to GO functional and KEGG pathway enrichment analyses. The fold changes in gene expression levels for glucose metabolism were further analyzed to evaluate expression differences in key enzyme genes， with four genes selected for validation via Real-time quantitative PCR. Result In the G2 vs G1 group，411 DEGs were identified （233 up-regulated，178 down-regulated）. In the S3 vs G2 group，3 664 DEGs were identified （1 799 up-regulated，1 865 down-regulated）.GO functional enrichment analysis revealed that DEGs in the G2 vs G1 group were primarily enriched in entries such as extracellular matrix and inflammatory and immune responses. Whereas the DEGs in the S3 vs G2 group were mainly enriched in entries related to immune system regulation and positive regulation of biological processes. KEGG pathway enrichment analysis indicated that no pathways related to glucose metabolism were significant enriched in the G2 vs G1 group. Whereas the S3 vs G2 group exhibited significant enrichment of DEGs across six glucose metabolism-related pathways， including glycolysis/gluconeogenesis， pyruvate metabolism， fructose and mannose metabolism， ascorbate and aldarate metabolism， pentose and glucuronate interconversions， and inositol phosphate metabolism. In the glycolysis pathway， the expression levels of HK1， HK2， HK3， and PFKP genes were significantly downregulated， while the expression level of PKLR gene was significantly upregulated （P<0.05）. In the citrate cycle pathway， the expression level of MDH2 gene was significantly increased （P<0.05）. The results of Real-time quantitative PCR validation for PCK1， G6PC1， IDH3B， and SDHC genes were consistent with those of transcriptomic sequencing. Conclusion The total alkaloids of Sophora alopecuroides could alleviate high-concentrate diet induced inflammation and fibrosis in sheep liver by enriching functions related to immune system regulation and positive regulation of biological processes. The expression of key gluconeogenesis enzyme genes was increased， the expression of upstream glycolysis enzyme genes was inhibited， and the expression of downstream glycolysis enzyme genes was promoted， while the liver citrate cycle was enhanced. Through these effects， the glucose metabolism disorders induced by high-concentrate diet were ameliorated， and systemic glucose metabolic homeostasis was maintained.

Key words: total alkaloids from Sophora alopecuroides; high-concentrate diet; sheep; glucose metabolism; transcriptome

CLC Number:

S816.7

SUN Lu, LI Shufang, WANG Lina, ZHAO Jianxin, LU Henan, WANG Hairong. Effects of Total Alkaloids from Sophora alopecuroides Added to a High-concentrate Diet on Hepatic Glucose Metabolism in Sheep Based on Transcriptome Sequencing[J]. China Animal Husbandry & Veterinary Medicine, 2026, 53(2): 671-681.

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References 38

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项目 Items	组别Groups
项目 Items	G1	G2/S3
原料Ingredients
混合牧草Mixed hay	50.00	30.00
玉米Corn	36.10	52.00
豆粕Soybean meal	12.00	15.20
石粉Limestone	0.60	1.00
磷酸氢钙CaHPO₄	0.10	0.10
食盐NaCl	0.50	0.50
预混料Premix^①	0.20	0.20
小苏打NaHCO₃	0.50	1.00
合计Total	100.00	100.00
营养成分Nutrient components^②
代谢能ME/（MJ/kg）	9.53	10.20
粗蛋白质CP	14.17	14.45
粗脂肪EE	2.84	3.72
中性洗涤纤维NDF	33.18	25.03
酸性洗涤纤维ADF	22.36	15.37
非结构性碳水化合物NSC	43.60	49.90
钙Ca	0.77	0.71
磷P	0.38	0.37

基因 Genes	引物序列 Primer sequences （5′→3′）	产物长度 Product length/bp	GenBank登录号 GenBank accession No.
PCK1	F：GCGGAGTACAAGGGCAAAGT	83	XM_004014441.5
	R：GCCAGGTACTGGCCAAAGTT
G6PC1	F：GGCATCAAACTCCTCTGGGT	76	XM_004012951.5
	R：GCTGTCCAAAGAGAATCCACTTAAA
IDH3B	F：GTTGCGGCATCTCAATCTCG	119	XM_004014386.6
	R：AGTCGGTTGTGGTGCTGTAG
SDHC	F：TGGGATCCGACACTTGATGT	107	XM_004002697.4
	R：GCCCTACAGAGGACAACACA
GAPDH	F：TCGGAGTGAACGGATTTGGC	76	NM_001190390.1
	R：ACGATGTCCACTTTGCCAGT

样品 Samples	原始数据 Raw reads/条	质控数据 Clean reads/条	Q20/%	Q30/%	GC 含量 GC content/%	总比对率 Total mapped/%	唯一比对率 Uniquely mapped/%
G1-1	48 466 846	47 954 082	98.04	94.12	48.97	96.48	84.97
G1-2	56 038 432	55 523 466	98.08	94.24	49.67	96.41	84.88
G1-3	48 483 846	48 016 986	98.07	94.24	50.16	96.67	86.77
G2-1	50 097 052	49 600 336	98.13	94.39	50.03	96.45	85.92
G2-2	52 449 996	51 964 618	98.10	94.27	49.28	96.82	84.83
G2-3	48 366 524	47 724 756	97.87	93.75	49.91	96.32	85.56
S3-1	50 859 634	50 391 610	97.28	95.35	50.31	96.89	84.90
S3-2	51 734 788	51 263 466	97.30	95.38	51.33	96.53	86.93
S3-3	49 821 332	49 302 912	97.40	95.54	48.96	97.01	79.20

基因Genes	通路 Pathway	G2 vs G1	S3 vs G2
PCK1	糖异生	0.76	2.26^*
PCK2		0.83	1.52^*
FBP1		0.97	1.87^*
G6PC1		0.92	3.50^*
PC		0.81	2.50^*
HK1	糖酵解	1.18	0.48^*
HK2		1.91	0.20^*
HK3		1.19	0.35^*
PFKP		1.17	0.43^*
PKLR		0.93	1.52^*
MDH2	柠檬酸循环	0.97	1.57^*
IDH3B		0.94	1.32
SDHC		1.01	1.12

Effects of Total Alkaloids from Sophora alopecuroides Added to a High-concentrate Diet on Hepatic Glucose Metabolism in Sheep Based on Transcriptome Sequencing

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