Chemical components and protective effects of Atractylodes japonica Koidz. ex Kitam against acetic acid-induced gastric ulcer in rats

Table 1 Sequences of the primers for real-time reverse transcription polymerase chain reaction

Gene	Forward primer	Reverse primer	Length
EGFR	CCTATGGGCCAAAGATCCCA	GAGGTTCCACGAGCTCTCTC	162 bp
ZO-1	CACCACAGACATCCAACCAG	CACCAACCACTCTCCCTTGT	230 bp
NF-κBp65	AGGCCATTGAAGTGATCCAG	CAGTGAGGGACTCCGAGAAG	204 bp
IκBα	CACGGAAGATGAGTTGCCCT	CAAGTCCACGTTCCTTTGGC	91 bp
GAPDH	AGACAGCCGCATCTTCTTGT	CTTGCCGTGGGTAGAGTCAT	207 bp

EGFR: Epidermal growth factor receptor; ZO-1: Zonula Occludens-1; NF-κB: Nuclear factor kappa-B; IκBα: IkappaBalpha.

Table 2 Identification of chemical constituents of Atractylodes japonica by UPLC Q-Exactive Orbitrap MS

No.	tR/min	Compound	Molecular formula	Adduct ion	Theoretical (m/z)	Measured (m/z)	Error (ppm)
1	0.83	Asparagine	C4H8N2O3	[M+H]+	133.06076	133.06033	-3.297
2	0.85	Sucrose	C12H22O11	[M−H+HCOOH]−	387.11441	387.11374	-1.741
3	0.86	Manninotriose	C18H32O16	[M−H]−	503.16092	503.16175	-1.665
4	0.85	Valine	C5H11NO2	[M+H]+	118.08625	118.08591	-2.923
5	0.87	5-hydroxymethylfurfural	C6H6O3	[M+H]+	127.03897	127.03864	-2.602
6	0.91	Niacinamide	C6H6N2O	[M+H]+	123.05529	123.05509	-1.620
7	1.19	Pyroglutamic acid	C5H7NO3	[M+H]+	130.04986	130.04953	-2.612
8	1.25	Tyrosine	C9H11NO3	[M+H]+	182.08116	182.08063	-2.964
9	1.27	Citric acid	C6H8O7	[M−H]−	191.01972	191.01880	-4.847
10	1.35	Isoleucine	C6H13NO2	[M+H]+	132.10190	132.10153	-2.840
11	1.46	Leucine	C6H13NO2	[M+H]+	132.10190	132.10153	-2.840
12	2.10	Phenylalanine	C9H11NO2	[M+H]+	166.08625	166.08577	-2.921
13	2.25	Neochlorogenic acid	C16H18O9	[M−H]−	353.08781	353.08783	0.050
14	2.44	Chlorogenic acid	C16H18O9	[M−H]−	353.08781	353.08752	-0.808
15	2.44	Atractyloside A	C21H36O10	[M−H+HCOOH]−	493.22905	493.22864	-0.831
16	2.80	Cryptochlorogenic acid	C16H18O9	[M−H]−	353.08781	353.08728	-1.488
17	2.86	Caffeic acid	C9H8O4	[M+H]+	181.04953	181.04890	-3.509
18	2.86	7-hydroxycoumarin	C9H6O3	[M+H]+	163.03897	163.03841	-3.439
19	3.17	Catechin	C15H14O6	[M−H]−	289.07176	289.07172	-0.143
20	3.19	Isochlorogenic acid B	C25H24O12	[M−H]−	515.11950	515.11890	-1.163
21	3.27	Isoorientin	C21H20O11	[M+H]+	449.10784	449.10669	-2.556
22	3.51	Rutin	C27H30O16	[M−H]−	609.14610	609.14630	0.315
23	3.52	Hyperoside	C21H20O12	[M−H]−	463.08819	463.08826	0.131
24	3.52	Quercetin	C15H10O7	[M+H]+	303.04992	303.04895	-3.528
25	3.68	Kaempferol-3-O- rutinoside	C27H30O15	[M−H]−	593.15119	593.15125	0.096
26	3.77	Isorhamnetin 3-O-neohesperidin	C28H32O16	[M−H]−	623.16176	623.16107	-1.104
27	3.78	Narcissoside	C28H32O16	[M+H]+	625.17631	625.17371	-4.161
28	3.78	Isorhamnetin	C16H12O7	[M+H]+	317.06558	317.06421	-4.318
29	3.83	Vanillin	C8H8O3	[M+H]+	153.05462	153.05406	-3.663
30	3.86	Isochlorogenic acid A	C25H24O12	[M−H]−	515.11950	515.11914	-0.697
31	3.96	Isochlorogenic acid C	C25H24O12	[M−H]−	515.11950	515.11896	-1.047
32	6.40	Palmitic acid	C16H32O2	[M+NH4]+	274.27406	274.27292	-4.141
33	6.47	Cinnamyl alcohol	C9H10O	[M+H]+	135.08044	135.07999	-3.343
34	6.51	Curcumenol	C15H22O2	[M+H]+	235.16925	235.16826	-4.237
35	7.34	Atractylenolide III	C15H20O3	[M+H]+	249.14852	249.14752	-4.018
36	7.62	Atractylenolactam	C15H19NO	[M+H]+	230.15394	230.15340	-2.350
37	9.24	3β-hydroxyatractylone	C15H20O2	[M+H]+	233.15361	233.15279	-3.502
38	9.55	Atractylenolide II	C15H20O2	[M+H]+	233.15361	233.15274	-3.716
39	12.00	Atractylenolide VI	C15H22	[M+H]+	203.17943	203.17889	-2.644
40	12.01	α-linolenic acid	C18H30O2	[M+H]+	279.23185	279.23108	-2.782
41	12.14	3β-acetoxyatracty	C17H22O3	[M+H]+	275.16417	275.16333	-3.057
42	12.38	Atractylenolide I	C15H18O2	[M+H]+	231.13796	231.13721	-3.229
43	12.95	Atractylenolide V	C15H20O4	[M+H]+	265.14343	265.14258	-3.227
44	13.42	(6E,12E)-tetradecadiene-8,10-diyne-1,3-diol diacetate	C18H22O4	[M+H]+	303.15908	303.15805	-3.416
45	13.97	β-elemene	C15H24	[M+H]+	205.19508	205.19443	-3.155
46	14.14	Eudesma-4(15),7(11)-dien-8-one	C15H22O	[M+H]+	219.17434	219.17346	-4.024
47	20.55	Atractylon	C15H20O	[M+H]+	217.15869	217.15800	-3.186
48	21.83	Biatractylolide	C30H38O4	[M+H]+	463.28428	463.28290	-2.992

Table 3 Identified metabolites of plasma

No.	Compound name	R.T. (min)	Formula	Exact mass (m/z)	HMDB ID	MG vs NG	P value	HA vs MG	P value
1	L-arginine	0.77	C6H14N4O2	174.1116	HMDB0000517	Decreased	< 0.01	Increased	< 0.05
2	Citrulline	0.79	C6H13N3O3	175.0956	HMDB0000904	Decreased	< 0.01	Increased	< 0.05
3	Taurine	0.79	C2H7NO3S	125.0146	HMDB0000251	Decreased	< 0.05	Increased	< 0.05
4	Adenine	1.42	C5H5N5	135.0545	HMDB0000034	Increased	< 0.05	Decreased	< 0.05
5	Glycocholic acid	6.31	C26H43NO6	465.3090	HMDB0000138	Decreased	< 0.05	Increased	< 0.05
6	Aldosterone	7.59	C21H28O5	360.1936	HMDB0000037	Decreased	< 0.05	Increased	< 0.05
7	Glycochenodeoxycholic acid	8.09	C26H43NO5	449.3138	HMDB0000637	Decreased	< 0.05	Increased	< 0.05
8	LysoPC (16:0/0:0)	10.17	C24H50NO7P	495.3324	HMDB0010382	Increased	< 0.01	Decreased	< 0.01
9	LysoPC (18:0/0:0)	11.20	C26H54NO7P	523.3637	HMDB0010384	Increased	< 0.05	Decreased	< 0.05
10	Eicosadienoic acid	13.76	C20H36O2	308.2715	HMDB0005060	Increased	< 0.01	Decreased	< 0.05

MG: Model group; NG: Normal group; HA: High dose group of the crude Atractylodes japonica.

Table 4 Results of enrichment analysis of biomarkers

No.	Pathway name	Match status	P value	-Log (P)	Impact
1	Primary bile acid biosynthesis	3/46	0.0012354	2.9082	0.0254
2	Arginine biosynthesis	2/14	0.0020576	2.6866	0.30457
3	Taurine and hypotaurine metabolism	1/8	0.040642	1.391	0.42857
4	Glycerophospholipid metabolism	1/36	0.17174	0.76513	0.01736
5	Arginine and proline metabolism	1/38	0.18047	0.74359	0.05786
6	Purine metabolism	1/65	0.29073	0.53651	0.00528
7	Steroid hormone biosynthesis	1/85	0.3638	0.43913	0.01032