Effects of Gut Microbiota on Bile Acid Profile and Bile Acid Metabolism in Piglets

doi:10.13560/j.cnki.biotech.bull.1985.2020-0269

Abstract

Abstract:

The aim of this study is to investigate the effects of gut microbiota on bile acid profile and bile acid metabolism in liver,jejunal contents and feces of pig models. Eleven piglets from aseptic caesarean section were randomly divided into germ-free（GF）group（5 piglets）and fecal microbiota transplantation（FMT）group（6 piglets）. GF group remained germ-free from beginning to end,and the FMT group was implanted with healthy porcine fecal bacteria at the age of 7 d. Liver,jejunal contents and fresh feces were collected for bile acid quantification at 42 d,as well as liver and jejunal tissues were collected for the quantification of genes related to bile acid metabolism. The results showed that：compared with the GF group,the total bile acid level in the piglet feces of the FMT group was significantly higher than that of the GF group（P<0.01）. The proportion of secondary bile acids in the jejunal contents and feces of the piglets in the FMT group to total bile acids was significantly higher than that in the GF group（P<0.05）,and the same trend was found in the liver（P<0.05）. Compared with the GF group,cholesterol 7-alpha hydroxy-lase gene（CYP7A1）,bile acid amino acid converting enzyme gene（BAT）,and farinol X receptor gene（FXR）,as well as genes related to bile acid synthesis and transport in the FMT group were down-regulated,while the expression of the intestinal transporter MRP2 gene was up-regulated. Thus,the transfer and colonization of gut microbiota can not only increase the bile acid excretion of piglets,increase the proportion of secondary bile acid,change the bile acid profile of piglets,but also affect the hepatointestinal circulation of bile acid.

Key words: germ-free pig, bile acid profile, bile acid metabolism

LI Meng-ying, ZHOU Hua, DING Yu-chun, LIU Zuo-hua, SUN Jing, LI Zhou-quan. Effects of Gut Microbiota on Bile Acid Profile and Bile Acid Metabolism in Piglets[J]. Biotechnology Bulletin, 2020, 36(10): 49-61.

Figures/Tables 11

References 35

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简称	英文名	中文名	结合/游离	初级/次级
TDCA	taurodeoxycholate	牛磺脱氧胆酸	T	S
TCA	taurocholic acid	牛磺胆酸	T	P
TCDCA	taurochenodeoxycholate	牛磺鹅脱氧胆酸	T	P
TLCA	taurolithocholate	牛磺石胆酸	T	S
TUDCA	tauroursodeoxycholic acid	牛磺熊脱氧胆酸	T	P
THDCA	taurohyodeoxycholic acid	牛磺猪脱氧胆酸	T	S
TαMCA	tauro α-muricholate	牛磺α鼠胆酸	T	S
TβMCA	tauro β-muricholate	牛磺β鼠胆酸	T	S
TωMCA	tauro ω-muricholate	牛磺ω鼠胆酸	T	S
THCA	taurohyocholate	牛磺猪胆酸	T	P
GCA	glycocholic acid	甘氨胆酸	G	P
GCDCA	glycochenodeoxycholate	甘氨鹅脱氧胆酸	G	P
GLCA	glycolithocholate	甘氨石胆酸	G	S
GUDCA	glycoursodeoxycholic acid	甘氨熊脱氧胆酸	G	S
GHDCA	glycohyodeoxycholate	甘氨猪脱氧胆酸	G	P
GHCA	glycohyocholate	甘氨猪胆酸	G	P
CDCA	chenodeoxycholic acid	鹅脱氧胆酸	U	P
isoLCA	isolithocholic acid	3β-羟基石胆酸	U	S
6-ketoLCA	6-ketolithocholic acid	6酮基石胆酸	U	S
7-ketoLCA	7-ketolithocholic acid	7酮基石胆酸	U	S
12-ketoLCA	12-ketolithocholic acid	12酮基石胆酸	U	S
6,7-diketoLCA	6,7-diketolithocholic acid	6,7-二酮石胆酸	U	S
7,12-diketoLCA	7,12-diketolithocholic acid	7,12-二酮石胆酸	U	S
dehydroLCA	dehydrolithocholic acid	去氢石胆酸	U	S
UDCA	ursodeoxycholic acid	熊脱氧胆酸	U	S
muroCA	murocholic acid	鼠胆酸	U	S
αMCA	α-muricholic acid	α鼠胆酸	U	S
βMCA	β-muricholic acid	β鼠胆酸	U	S
ωMCA	ω-muricholic acid	ω鼠胆酸	U	S
HCA	γ-muricholic acid\hyocholic acid	猪胆酸	U	P
12-DHCA	12-dehydrocholic acid	12-去氢胆酸	U	S
3-DHCA	3-dehydrocholic acid	3-去氢胆酸	U	S
apoCA	apocholic acid	原胆酸	U	S
NorDCA	23-nordeoxycholic acid	23-脱甲脱氧胆酸	U	S
CDCA-24G	Chenodeoxycholic acid 24-Acyl-β-D-glucuronide	鹅去氧胆酸-24-酰基-β-D-葡萄糖醛酸	U	P
UCA	ursocholic acid	熊果胆酸	U	S
βUDCA	3β-Ursodeoxycholic Acid	3β-熊脱氧胆酸	U	S
βHDCA	β-Hyodeoxycholic Acid	β-猪脱氧胆酸	U	S
βCA	3β-Cholic Acid	3β-胆酸	U	S
ACA	Allocholic acid	别胆酸	U	S
NorCA	Nor Cholic acid	正胆酸	U	S
CDCA-3G	Chenodeoxycholic acid-3-β-D-glucuronide	鹅去氧胆酸-3-β-D-葡萄糖醛酸	U	P
βUCA	β-ursocholic acid	β-熊果胆酸	U	S
βCDCA	β-chenodeoxycholic acid	β-鹅脱氧胆酸	U	P
βDCA	β-deoxycholic acid	β-脱氧胆酸	U	S
7-DHCA	7-dehydrocholic acid	7-去氢胆酸	U	S
DCA	deoxycholic acid	脱氧胆酸	U	S
CA	cholic acid	胆酸	U	P
LCA	lithocholic acid	石胆酸	U	S
HDCA	α-hyodeoxycholic acid	猪脱氧胆酸	U	P

简称	英文名	中文名	结合/游离	初级/次级
TDCA	taurodeoxycholate	牛磺脱氧胆酸	T	S
TCA	taurocholic acid	牛磺胆酸	T	P
TCDCA	taurochenodeoxycholate	牛磺鹅脱氧胆酸	T	P
TLCA	taurolithocholate	牛磺石胆酸	T	S
TUDCA	tauroursodeoxycholic acid	牛磺熊脱氧胆酸	T	P
THDCA	taurohyodeoxycholic acid	牛磺猪脱氧胆酸	T	S
TαMCA	tauro α-muricholate	牛磺α鼠胆酸	T	S
TβMCA	tauro β-muricholate	牛磺β鼠胆酸	T	S
TωMCA	tauro ω-muricholate	牛磺ω鼠胆酸	T	S
THCA	taurohyocholate	牛磺猪胆酸	T	P
GCA	glycocholic acid	甘氨胆酸	G	P
GCDCA	glycochenodeoxycholate	甘氨鹅脱氧胆酸	G	P
GLCA	glycolithocholate	甘氨石胆酸	G	S
GUDCA	glycoursodeoxycholic acid	甘氨熊脱氧胆酸	G	S
GHDCA	glycohyodeoxycholate	甘氨猪脱氧胆酸	G	P
GHCA	glycohyocholate	甘氨猪胆酸	G	P
CDCA	chenodeoxycholic acid	鹅脱氧胆酸	U	P
isoLCA	isolithocholic acid	3β-羟基石胆酸	U	S
6-ketoLCA	6-ketolithocholic acid	6酮基石胆酸	U	S
7-ketoLCA	7-ketolithocholic acid	7酮基石胆酸	U	S
12-ketoLCA	12-ketolithocholic acid	12酮基石胆酸	U	S
6,7-diketoLCA	6,7-diketolithocholic acid	6,7-二酮石胆酸	U	S
7,12-diketoLCA	7,12-diketolithocholic acid	7,12-二酮石胆酸	U	S
dehydroLCA	dehydrolithocholic acid	去氢石胆酸	U	S
UDCA	ursodeoxycholic acid	熊脱氧胆酸	U	S
muroCA	murocholic acid	鼠胆酸	U	S
αMCA	α-muricholic acid	α鼠胆酸	U	S
βMCA	β-muricholic acid	β鼠胆酸	U	S
ωMCA	ω-muricholic acid	ω鼠胆酸	U	S
HCA	γ-muricholic acid\hyocholic acid	猪胆酸	U	P
12-DHCA	12-dehydrocholic acid	12-去氢胆酸	U	S
3-DHCA	3-dehydrocholic acid	3-去氢胆酸	U	S
apoCA	apocholic acid	原胆酸	U	S
NorDCA	23-nordeoxycholic acid	23-脱甲脱氧胆酸	U	S
CDCA-24G	Chenodeoxycholic acid 24-Acyl-β-D-glucuronide	鹅去氧胆酸-24-酰基-β-D-葡萄糖醛酸	U	P
UCA	ursocholic acid	熊果胆酸	U	S
βUDCA	3β-Ursodeoxycholic Acid	3β-熊脱氧胆酸	U	S
βHDCA	β-Hyodeoxycholic Acid	β-猪脱氧胆酸	U	S
βCA	3β-Cholic Acid	3β-胆酸	U	S
ACA	Allocholic acid	别胆酸	U	S
NorCA	Nor Cholic acid	正胆酸	U	S
CDCA-3G	Chenodeoxycholic acid-3-β-D-glucuronide	鹅去氧胆酸-3-β-D-葡萄糖醛酸	U	P
βUCA	β-ursocholic acid	β-熊果胆酸	U	S
βCDCA	β-chenodeoxycholic acid	β-鹅脱氧胆酸	U	P
βDCA	β-deoxycholic acid	β-脱氧胆酸	U	S
7-DHCA	7-dehydrocholic acid	7-去氢胆酸	U	S
DCA	deoxycholic acid	脱氧胆酸	U	S
CA	cholic acid	胆酸	U	P
LCA	lithocholic acid	石胆酸	U	S
HDCA	α-hyodeoxycholic acid	猪脱氧胆酸	U	P

饲粮组成/%	含量	营养水平
普通玉米,CP 7.8%	14.10	CP,%	19.00
膨化玉米,CP 7.8%	10.00	DE,Kcal/kg	3 622.00
去皮豆粕,CP 47.13%	13.65	NE,Kcal/kg	2 645.00
膨化大豆,CP 35.5%	7.00	SID Lys,%	1.23
大豆浓缩蛋白,CP 65.2%	5.00	SID Met,%	0.36
乳清粉,CP 3.26%	5.00	SID Thr,%	0.73
鱼粉,65.2%	3.00	SID Trp,%	0.20
血浆蛋白粉,CP 76.66%	3.00	Ca,%	0.70
葡萄糖	2.00	P,%	0.60
大豆油	1.40	AP,%	0.45
玉米淀粉	29.70
纤维	3.00
石灰石	0.45
磷酸氢钙	1.25
L-Lys（78%）	0.29
DL-Met（99%）	0.12
L-Thr（98.5%）	0.13
L-Trp（98%）	0.01
维生素-矿物质预混料	0.50
氯化钠	0.25
氯化胆碱	0.15
总计	100.00

饲粮组成/%	含量	营养水平
普通玉米,CP 7.8%	14.10	CP,%	19.00
膨化玉米,CP 7.8%	10.00	DE,Kcal/kg	3 622.00
去皮豆粕,CP 47.13%	13.65	NE,Kcal/kg	2 645.00
膨化大豆,CP 35.5%	7.00	SID Lys,%	1.23
大豆浓缩蛋白,CP 65.2%	5.00	SID Met,%	0.36
乳清粉,CP 3.26%	5.00	SID Thr,%	0.73
鱼粉,65.2%	3.00	SID Trp,%	0.20
血浆蛋白粉,CP 76.66%	3.00	Ca,%	0.70
葡萄糖	2.00	P,%	0.60
大豆油	1.40	AP,%	0.45
玉米淀粉	29.70
纤维	3.00
石灰石	0.45
磷酸氢钙	1.25
L-Lys（78%）	0.29
DL-Met（99%）	0.12
L-Thr（98.5%）	0.13
L-Trp（98%）	0.01
维生素-矿物质预混料	0.50
氯化钠	0.25
氯化胆碱	0.15
总计	100.00

基因	上游引物（5'-3'）	下游引物（5'-3'）	产物长度/bp	参考文献
GAPDH	CCAGGGCTGCTTTTAACTCTG	GTGGGTGGAATCATACTGGAACAT	104	[16]
CYP7A1	GAAAGAGAGACCACATCTCGG	GAATGGTGTTGGCTTGCGAT	123	[17]
CYP4A21	GATCCAGGAACTGCATTGGGA	CACAATTCCTTGAATAGGAACGGG	128
CYP27A1	ACTGAAGACCGCGATGAAAC	CAAAGGCGAATCAGGAAGGG	106	[17]
FXR	TATGAACTCAGGCGAATGCCTGCT	ATCCAGATGCTCTGTCTCCGCAAA	154	[17]
LRH1	GGTACCACTATGGGCTCCTCAC	TCGGCCCTTACCGCTTCT	193	[18]
OATP1	TGCTTAAAACTCCCACCCCT	GTTTGCTTCATTCACGGCTT	120	[19]
BESP	CGCAGCGTGAAGAAATGTGG	AAAACCGAAACAGTTGAAAGAGGC	136
FGFR4	GCTCAGAGGTGGAGGTCCTA	GCCTGCCAGACAGGTGTATT	74	[17]
BAT	GGCTGATGATCCGAGAAGGG	ATGCCCCCAAACAAGTCGAT	94	[17]
KLβ	GCACCGAGTGGAAGGAGT	TTGCCAGTAGGAAGGATTG	150	[17]
MRP2	GAACAGGTTTGCTGGCGATATT	GCCAGGAGCGCAAAGACA	65	[17]
SHP	ACCATTCTCTTCAACCCTGATGT	GCTGCGAGGAGAACACGAG	149
HNF4α1	GCCTCTCCAAAACCCTGGTC	CTGATGGGGATGTGTCATTGC	126
BACS	CTGGCTCCCTGCCTATGCT	GAACGTGCTTGTGGTCTCCAA	70	[17]
NTCP	TGTCATCAAGGGAGGAACGA	CGAGCATTGAGGCGGAAAAG	189
ASBT	AAGTTCCTGGGGCACGTAAA	CTCCTGGACAGCATCCCATT	161	[17]
OSTα	TGTACAAGAACACTCGCTGC	GAACACACACACTATCGTGGG	80	[17]
FGF19	AAGATGCAAGGGCAGACTCA	AGATGGTGTTTCTTGGACCAGT	101
IBABP	GCAAGGAGTGCGACATAGAGAC	TGGTGGTAGTTGGGGCTGTT	100
MRP3	TGGACAAAGGGACAATAGCTGAGT	TGGCCATCCCGTAGAAGATG	78	[17]
TGR5	TGCTGTCCCTCATCTCATTGG	TGTGTAGCGATGATCACCCAG	80	[17]