3, 3-二甲基-1-丁醇抑制肠道三甲胺生成可缓解溃疡性结肠炎及继发性肝损伤

王鑫楠; 刘丽芳

doi:10.11665/j.issn.1000-5048.2023080801

3, 3-二甲基-1-丁醇抑制肠道三甲胺生成可缓解溃疡性结肠炎及继发性肝损伤

王鑫楠^{1, 2,},
刘丽芳^1, ,

1.
中国药科大学中药学院生药学系, 南京211198
2.
江苏省疾病预防控制中心, 南京210009

基金项目: 国家自然科学基金项目（No.82074089）

详细信息

通讯作者:
刘丽芳: Tel：13813941057　E-mail：liulifang69@126.com

中图分类号: R965
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出版历程
- 收稿日期: 2023-08-07
- 刊出日期: 2024-04-24

3, 3-Dimethyl-1-butanol attenuates ulcerative colitis and secondary liver injury by reducing trimethylamine production

WANG Xinnan^{1, 2,},
LIU Lifang^1, ,

1.
Department of Pharmacognosy, School of Traditional Chinese Pharmacy, China Pharmaceutical University, Nanjing 211198
2.
Jiangsu Provincial Center for Disease Control and Prevention, Nanjing 210009, China

Funds: This study was supported by the National Natural Science Foundation of China (No. 82074089)

摘要

摘要:
溃疡性结肠炎（ulcerative colitis，UC）是多种因素导致的肠道慢性疾病，严重的肠炎会引起肝脏损伤。肠道炎症诱发菌群紊乱，导致肠道中过多的胆碱转化为三甲胺；宿主中过低的胆碱生物利用度是造成肝脏损伤的重要原因。3，3-二甲基-1-丁醇（3,3-dimethyl-1-butanol，DMB）作为胆碱的结构类似物，可有效抑制肠道中胆碱转化为三甲胺，本研究旨在探究DMB能否通过减少肠道内的胆碱的不良转化来改善UC小鼠结肠炎症及继发性肝损伤。采用葡聚糖硫酸钠诱导的结肠炎模型，造模结束后，评估结肠及肝组织病理，检测肝功能相关生化指标，使用超高效液相色谱串联质谱法检测UC所致的体内胆碱代谢变化。结果表明，DMB能够减轻UC小鼠体重下降指数，缓解结肠炎症，减少肝脏损伤，对小鼠的血清、肠道内容物与肝脏进行胆碱相关代谢物的检测，发现DMB能够有效抑制肠道中胆碱转化为三甲胺，提高宿主的胆碱利用度，有效缓解结肠炎的恶化，从而减少由于严重的肠道病变导致的肝脏受损。
- 胆碱代谢 /
- 结肠炎 /
- 肠道菌群代谢 /
- 三甲胺
Abstract:
Ulcerative colitis (UC) is a chronic intestinal disease caused by a variety of factors. Severe intestinal inflammation can also cause liver injury. Based on the previous research, microbial dysbiosis in the inflammatory state leads to the conversion of excess choline into trimethylamine (TMA) by the intestinal flora, which competes with the host for the use of the nutrient choline, and induces liver injury. 3, 3-dimethyl-1-butanol (DMB), a structural analogue of choline, can reduce TMA levels from choline conversion. The aim of this study was to investigate the protective effect and possible mechanism of DMB on UC and secondary liver injury. Dextran sulfate sodium-induced acute colitis model in mice was established. The weight of mice, and collected serum, liver and intestinal contents after mice sacrifice were measured. The morphological changes of colon and liver were observed; liver function was detected with the kit of biochemical indexes; UHPLC-MS/MS was applied to detect changes in choline metabolism in vivo. The experimental results showed that DMB could attenuate body weight loss index, improve colonic inflammation, and reduce liver injury in UC mice. The detection of choline-related metabolites in serum, intestinal contents and liver showed that DMB could effectively inhibit the production of trimethylamine in the intestine, improve the availability of host choline, effectively alleviate colitis deterioration, and reduce liver damage caused by severe intestinal lesions.
- choline metabolism /
- colitis /
- gut microbiota metabolism /
- trimethylamine

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Figure 1. Pathways for the choline metabolism; 3,3-dimethyl-1-butanol(DMB) suppress trimethylamine(TMA) levels from choline conversion

ChAT:Choline acetyltransferase;CHDH:Choline dehydrogenase;CK:Choline kinase;CPT:Choline phosphotransferase;CT:Phosphocholine cytidylytransferase; FMO3:Flavin monooxygenase; PLD:Phospholipase D

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Figure 2. Influence of oral DMB on colitis mice

A: Percentage change chart of body weight; B: Mean colon length of each group; C: Representative images in each group; D: DAI score of each group (^*P < 0.05, ^**P < 0.01, ^***P < 0.001 vs control group; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 vs model group; n ≥ 6 )

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Figure 3. Hematoxylin and eosin staining of colon tissues and histological score in each group

^*P < 0.05, ^**P < 0.01, ^***P < 0.001 vs control group; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 vs model group; n ≥ 6 per group

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Figure 4. DMB intervention ameliorated secondary liver injury

A: Hematoxylin and eosin staining of liver tissues of each group; B: Alanine transaminase(ALT), aspartate aminotransferase(AST) and alkaline phosphatase(AKP) activity in plasma^*P < 0.05, ^**P < 0.01, ^***P < 0.001 vs control group; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001, vs model group; n ≥ 6 per group

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Figure 5. Bar graph of choline metabolism-associated metabolites quantification

A: Intestinal contents; B: Liver; C: Serum ^*P < 0.05, ^**P < 0.01, ^***P < 0.001 vs control group; ^#P < 0.05, ^##P < 0.01, ^###P < 0.001 vs model group; n ≥ 6 per group

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Table 1 MRM transition parameters, LLOD, LLOQ, and RSD for the targeted analytes in choline metabolism

Analyte	MRM	DP^a	CE^b	Sensitivity
Analyte	MRM	DP^a	CE^b	LLOD/ (ng/mL)	RSD	LLOQ/ (ng/mL)	RSD
TMA	60.1>45.1	25	20	2	16.95	5	10.34
TMAO	75.9>59.1	40	30	0.5	16.39	2	7.74
CHO	104.0>45.0	100	35	0.005	19.05	0.05	14.24
CHO-D9 (IS)	113.1>69.0	100	35
ACH	147.2>88.0	70	20	0.05	14.69	0.1	14.91
Bet	118.1>59.1	120	30	0.1	8.46	0.5	7.97
PC	758.2>86.0	130	40	5	14.56	10	12.66
DMPC (IS)	678.4>86.0	130	40
a:Declustering potential;b:Collision energy TMA: Trimethylamine; TMAO: Trimethylamine N-oxide; CHO: Choline; Bet: Betaine; PC: Phosphatidylcholine; ACH: Acetylcholine; CHO-D9: Chloride-(trimethy-d9) choline; DMPC: 1 2-Dimyristoyl-sn-glycero-3-phosphocholine

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Table 2 Solvent-only and matrix-matched calibration curves and R² of samples

Compd.	Solvent-only		Matrix-matched		Dynamic range/ (ng/mL)
Compd.	Calibration curves	R²	Calibration curves	R²	Dynamic range/ (ng/mL)
Intestinal contents
TMA	y = 0.0017x + 0.0057	0.995 3	y = 0.0015x + 0.0349	0.998 0	3.75−375
TMAO	y = 0.0143x + 0.037	0.998 9	y = 0.0130x + 0.0393	0.998 0	1.25−125
CHO	y = 0.0115x – 0.0073	0.999 7	y = 0.0112x + 1.9279	0.9978	5−500
ACH	y = 0.0028x – 0.001	0.998 0	y = 0.0027x + 0.0025	0.9931	0.5−50
Bet	y = 0.0383x + 0.0576	0.9993	y = 0.0343x + 0.9758	0.999 0	1−100
PC	y = 0.7703x + 0.0217	0.9975	y = 0.5176x + 1.1683	0.9939	12.5−12500
Liver
TMA	y = 0.0095x + 0.0038	0.9997	y = 0.0089x + 0.1968	0.9972	1.25−125
TMAO	y = 0.0162x + 0.0402	0.9988	y = 0.0150x + 0.0329	0.9988	1.25−125
CHO	y = 0.0116x + 0.0227	0.9999	y = 0.0118x + 1.8986	0.9976	10−1000
ACH	y = 0.0013x – 0.0007	0.9992	y = 0.0013x + 0.0021	0.9937	1.25−125
Bet	y = 0.0181x + 0.0481	0.9993	y = 0.0176x + 5.1867	0.9955	4−400
PC	y = 0.4209x + 0.1404	0.9943	y = 0.3533x + 11.624	0.9918	300−30000
Serum
TMA	y = 0.0057x + 0.0197	0.9955	y = 0.0053x + 0.0192	0.998 0	1.8−180
TMAO	y = 0.0159x + 0.0761	0.9982	y = 0.0150x + 0.0502	0.9998	2.25−225
CHO	y = 0.0129x + 0.0068	0.9999	y = 0.0134x + 0.9229	0.9995	6.75−675
ACH	y = 0.0028x – 0.0006	0.9981	y = 0.0028x + 0.0005	0.998 0	0.9−90
Bet	y = 0.0395x + 0.1593	0.9986	y = 0.0371x + 0.8011	0.9997	4.5−450
PC	y = 0.0006x + 0.0775	0.995 0	y = 0.0004x + 3.3674	0.9967	90−9000

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Table 3 Methodological observations including precision and accuracy, recovery and matrix effects results

Analyte	Added/ (ng/mL)	Matrix effect	Recovery		Intraday		Interday
Analyte	Added/ (ng/mL)	t-value	Mean/%	RSD/%	Accuracy/%	Precision/%	Accuracy/%	Precision/%
Intestinal contents
	15		101.01	8.87	−6.23	6.60	−3.61	14.45
TMA	112.5	0.270 3	100.50	4.12	9.05	5.27	8.29	5.21
	187.5		105.43	6.29	9.60	7.95	10.40	9.62
	5		90.90	10.79	4.12	6.33	2.30	6.03
TMAO	37.5	0.1574	102.91	4.94	7.61	7.39	6.44	6.95
	62.5		105.53	7.33	3.58	2.92	4.43	4.52
	20		103.49	6.02	−9.57	3.31	−6.62	2.92
CHO	150	0.0221	97.78	9.96	−8.75	4.37	−9.17	3.24
	250		90.76	6.10	−2.45	4.87	−5.47	5.08
	2		105.14	6.73	−1.71	13.33	5.14	4.67
ACH	15	0.0493	101.74	5.70	1.74	5.33	1.74	5.37
	25		116.08	4.01	18.63	3.69	17.33	4.45
	4		89.97	9.39	−17.39	2.62	−14.80	2.64
BET	30	0.2426	97.01	4.44	−1.92	1.76	1.97	3.97
	50		101.98	6.18	3.11	4.42	−2.73	2.17
	50		95.34	9.21	14.74	7.99	−9.03	8.34
PC	375	0.3543	86.83	6.45	−0.83	8.41	4.89	8.25
	625		108.79	9.52	−5.41	3.40	−12.20	2.45
Liver
	5		102.47	10.80	−18.76	8.67	−9.70	5.82
TMA	37.5	0.1256	89.42	7.39	−1.78	5.43	−0.49	4.72
	62.5		100.64	5.70	−13.82	12.16	−16.83	11.10
	5		97.05	7.78	7.09	5.16	8.26	4.79
TMAO	37.5	0.1275	96.94	4.75	−3.98	5.12	−2.98	4.21
	62.5		93.64	3.49	−3.90	3.65	−4.63	3.05
	40		112.08	5.20	17.38	6.02	11.81	2.68
CHO	300	0.0248	101.53	2.61	−5.49	1.76	−5.32	1.80
	500		101.44	2.09	−9.76	1.80	−9.39	1.71
	5		114.08	6.74	17.77	7.92	14.91	5.56
ACH	37.5	0.0160	98.90	9.06	−2.11	8.36	−1.26	8.33
	2		96.19	5.70	−7.23	12.01	−6.09	12.78
	16		87.37	8.87	4.21	3.15	13.73	3.27
BET	120	0.0392	102.23	5.56	−4.06	3.93	−5.54	3.65
	200		94.45	9.36	−2.06	3.01	2.23	1.63
	1200		104.40	11.21	−8.68	11.10	−14.11	11.17
PC	9000	0.1895	89.23	6.76	0.25	9.22	−8.62	8.02
	15000		100.80	13.85	7.64	6.81	0.97	6.05
Serum
	7.2		80.61	9.82	−8.49	7.33	−11.82	4.36
TMA	54	0.1574	89.53	1.90	−8.46	1.73	−8.36	1.91
	90		96.75	4.49	−2.05	4.26	−0.62	2.45
	9		94.67	6.13	−4.87	4.39	−3.41	3.55
TMAO	67.5	0.2001	97.22	2.93	−2.71	2.79	−3.37	2.42
	112.5		99.08	1.48	−0.87	1.44	−0.75	1.53
	27		90.24	12.61	−9.60	5.87	7.13	10.52
CHO	202.5	0.0580	97.89	2.75	−0.67	2.02	0.17	1.21
	337.5		99.75	2.07	0.60	1.70	0.85	1.76
	3.6		82.76	10.97	−17.24	10.35	−17.42	7.98
ACH	27	0.0000	97.06	3.61	−2.93	3.59	−4.00	2.26
	45		97.24	5.02	−2.75	5.00	−2.86	5.16
	18		95.90	5.94	−4.09	2.63	−3.40	2.73
BET	135	0.1404	99.76	2.40	−0.23	2.07	−0.49	2.17
	225		101.85	2.44	1.85	2.23	1.77	2.44
	360		86.66	4.24	−10.91	3.58	4.86	3.27
PC	2700	0.5438	95.69	9.71	0.03	2.25	2.25	1.90
	4500		111.36	11.95	14.75	5.98	18.58	5.36

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参考文献(24)

[1]	Ordas I, Eckmann L, Talamini M, et al. Ulcerative colitis[J]. Lancet, 2012, 380(9853): 1606-1619. doi: 10.1016/S0140-6736(12)60150-0
[2]	Al-Horani R, Spanudakis E, Hamad B. The market for ulcerative colitis[J]. Nat Rev Drug Discov, 2022, 21(1): 15-16. doi: 10.1038/d41573-021-00194-5
[3]	Knight C, Murray KF. Hepatobiliary associations with inflammatory bowel disease[J]. Expert Rev Gastroenterol Hepatol, 2009, 3(6): 681-691. doi: 10.1586/egh.09.53
[4]	Henao-Mejia J, Elinav E, Jin C, et al. Inflammasome-mediated dysbiosis regulates progression of NAFLD and obesity[J]. Nature, 2012, 482(7384): 179-185. doi: 10.1038/nature10809
[5]	Spencer MD, Hamp TJ, Reid RW, et al. Association between composition of the human gastrointestinal microbiome and development of fatty liver with choline deficiency[J]. Gastroenterology, 2011, 140(3): 976-986. doi: 10.1053/j.gastro.2010.11.049
[6]	Pogribny IP, Beland FA. DNA hypomethylation in the origin and pathogenesis of human diseases[J]. Cell Mol Life Sci, 2009, 66(14): 2249-2261. doi: 10.1007/s00018-009-0015-5
[7]	Glunde K, Bhujwalla ZM, Ronen SM. Choline metabolism in malignant transformation[J]. Nat Rev Cancer, 2011, 11: 835-848. doi: 10.1038/nrc3162
[8]	Chittim CL, Martinez Del CA, Balskus EP. Gut bacterial phospholipase Ds support disease-associated metabolism by generating choline[J]. Nat Microbiol, 2019, 4(1): 155-163.
[9]	Teodoro JS, Rolo AP, Duarte FV, et al. Differential alterations in mitochondrial function induced by a choline-deficient diet: understanding fatty liver disease progression[J]. Mitochondrion, 2008, 8(5/6): 367-376.
[10]	Zeisel SH. Dietary choline deficiency causes DNA strand breaks and alters epigenetic marks on DNA and histones[J]. Mutat Res, 2012, 733(1/2): 34-38.
[11]	Hosseinkhani F, Heinken A, Thiele I, et al. The contribution of gut bacterial metabolites in the human immune signaling pathway of non-communicable diseases[J]. Gut Microbes, 2021, 13(1): 1-22.
[12]	Wang Z, Roberts AB, Buffa JA, et al. Non-lethal inhibition of gut microbial trimethylamine production for the treatment of atherosclerosis[J]. Cell, 2015, 163(7): 1585-1595. doi: 10.1016/j.cell.2015.11.055
[13]	Hamamoto N, Maemura K, Hirata I, et al. Inhibition of dextran sulphate sodium (DSS) -induced colitis in mice by intra colonically administered antibodies against adhesion molecules (endothelial leucocyte adhesion molecule-1 (ELAM-1) or intercellular adhesion molecule-1 (ICAM-1))[J]. Clin Exp Immunol, 1999, 117(3): 462-468.
[14]	Morris GP, Beck PL, Herridge MS, et al. Hapten-induced model of chronic inflammation and ulceration in the rat colon[J]. Gastroenterology, 1989, 96(3): 795-803. doi: 10.1016/0016-5085(89)90904-9
[15]	Wang XN, Liu JQ, Shi ZQ, et al. Orthogonal label and label-free dual pretreatment for targeted profiling of neurotransmitters in enteric nervous system[J]. Anal Chim Acta, 2020, 1139: 68-78. doi: 10.1016/j.aca.2020.09.031
[16]	Zheng J, Mandal R, Wishart DS. A sensitive, high-throughput LC-MS/MS method for measuring catecholamines in low volume serum[J]. Anal Chim Acta, 2018, 1037: 159-167. doi: 10.1016/j.aca.2018.01.021
[17]	Goh YQ, Cheam G, Wang Y. Understanding choline bioavailability and utilization: first step toward personalizing choline nutrition[J]. J Agric Food Chem, 2021, 69(37): 10774-10789. doi: 10.1021/acs.jafc.1c03077
[18]	Stremmel W, Merle U, Zahn A, et al. Retarded release phosphatidylcholine benefits patients with chronic active ulcerative colitis[J]. Gut, 2005, 54(7): 966-971. doi: 10.1136/gut.2004.052316
[19]	Corbin KD, Zeisel SH. Choline metabolism provides novel insights into nonalcoholic fatty liver disease and its progression[J]. Curr Opin Gastroenterol, 2012, 28(2): 159-165. doi: 10.1097/MOG.0b013e32834e7b4b
[20]	Wang G, Kong B, Shuai W, et al. 3, 3-Dimethyl-1-butanol attenuates cardiac remodeling in pressure-overload-induced heart failure mice[J]. J Nutr Biochem, 2020, 78: 108341. doi: 10.1016/j.jnutbio.2020.108341
[21]	Hsu CN, Chang-Chien GP, Lin S, et al. Targeting on gut microbial metabolite trimethylamine-n-oxide and short-chain fatty acid to prevent maternal high-fructose-diet-induced developmental programming of hypertension in adult male offspring[J]. Mol Nutr Food Res, 2019, 63(18): e1900073. doi: 10.1002/mnfr.201900073
[22]	González-Correa C, Moleón J, Miñano S, et al. Trimethylamine n-oxide promotes autoimmunity and a loss of vascular function in toll-like receptor 7-driven lupus mice[J]. Antioxidants (Basel), 2021, 11(1): 84. doi: 10.3390/antiox11010084
[23]	Romano KA, Martinez-Del Campo A, Kasahara K, et al. Metabolic, epigenetic, and trans generational effects of gut bacterial choline consumption[J]. Cell Host Microb, 2017, 22(3): 279-290. doi: 10.1016/j.chom.2017.07.021
[24]	Randrianarisoa E, Lehn-Stefan A, Wang X, et al. Relationship of serum trimethylamine n-oxide (tmao) levels with early atherosclerosis in humans[J]. Sci Rep, 2016, 27(6): 26745.

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