Qushi Huoxue ointment ameliorates metabolic associated steatotic liver disease through autophagy activation and ferroptosis inhibition

Table 1 Quantitative real-time polymerase chain reaction primer sequence list

Gene	Primer	Sequence
Beclin1	Forward	CTGTGGAGTGGAATGAAA
	Reverse	TAGGGAACAAGTCGGTAC
P62	Forward	ATTTCCTGAAGAATGTGGG
	Reverse	CTGCTTGGCTGAGTGTTA
Slc7a11	Forward	TTGGAGCCCTGTCCTATGC
	Reverse	CGAGCAGTTCCACCCAGAC
GPX4	Forward	ATTCTCAGCCAAGGACAT
	Reverse	CAGGATTCGTAAACCACA

GPX4: Glutathione peroxidase 4.

Table 2 Qualitative results of blood ingredients in Qushi Huoxue ointment

No.	Compound	RT/minute	Formula	Reference ion	m/z	Annotation MW	Calc. MW	Mass error (ppm)	Sort
1	L(-)-carnitine[24]	1.253	C7H15NO3	[M+H]+1	162.11258	161.10519	161.10532	0.00013	Amino acid derivatives
2	Emodin[25]	16.835	C15H10O5	[M+H]+1	271.06018	270.05282	270.0529	0.00007	Anthraquinones
3	Trigonelline[26]	1.402	C7H7NO2	[M+H]+1	138.05501	137.04768	137.04773	0.00005	Quaternary ammonium alkaloid
4	1-heptanesulfonic acid	21.624	C7H16O3S	[M-H]-1	179.07375	180.08201	180.08102	-0.00099	Alkylsulfonic acid
5	18-β-glycyrrhetinic acid[27]	21.286	C30H46O4	[M-H]-1	469.33200	470.33961	470.33928	-0.00033	Triterpenes
6	Cyclo (leucylprolyl)[28]	11.266	C11H18N2O2	[M+H]+1	211.14432	210.13683	210.13704	0.00021	Diketopiperazines
7	DL-homoserine[29]	1.307	C4H9NO3	[M+H]+1	120.06588	119.05824	119.0586	0.00036	Non-proteinogenic amino acids
8	2,3,4,9-tetrahydro-1H-β-carboline-3-carboxylic acid[30]	8.669	C12H12N2O2	[M+H]+1	217.09747	216.08988	216.0902	0.00032	β-carboline alkaloids
9	L-histidine[31]	1.183	C6H9N3O2	[M-H]-1	154.06104	155.06948	155.06831	-0.00116	Proteinogenic amino acids
10	cis-resveratrol[32]	11.887	C14H12O3	[M+H]+1	229.08614	228.07864	228.07886	0.00022	Stilbenes
11	Glycitein[33]	15.457	C16H12O5	[M+H]+1	285.07593	284.06847	284.0686	0.00012	Isoflavones
12	4-oxoproline[34]	2.715	C5H7NO3	[M-H]-1	128.03416	129.04259	129.04144	-0.00115	Amino acid
13	N5-(1,3,5-trimethyl-1H-pyrazol-4-yl)-1H-1,2,4-triazole-3,5-diamine	6.652	C8H13N7	[M+H]+1	208.13333	207.12324	207.12605	0.00281	Heterocyclic compounds
14	2-phenoxypropanoic acid	9.34	C9H10O3	[M-H]-1	165.05479	166.06299	166.06207	-0.00092	Carboxylic acids
15	Caffeic acid[35]	18.577	C9H8O4	[M+H]+1	149.02333	180.04226	148.01604	-32.02622	Phenolic acid
16	(3S,3aR,4S,4aR,7aR,8R,9aR)-3,4a,8-trimethyl-2,5-dioxo-2H,3H,9aH-azuleno[6,5-b]yl(2Z)-2-methylbut-2-enoate]	18.415	C20H26O5	[M+H]+1	369.17236	346.17802	368.16509	21.98706	Esters
17	RKK	20.151	C18H38N8O4	[M+H]+1	431.31555	430.3016	430.30821	0.00661	Amino acids
18	(3R,4S)-4,6,8-Trihydroxy-7-methoxy-3-methyl-3,4-dihydro-1H-isochromen-1-one	10.277	C11H12O6	[M-H]-1	239.05597	240.06339	240.06325	-0.00014	Isocoumarin derivatives
19	Apigetrin[36]	12.243	C21H20O10	[M-H]-1	431.09830	432.10565	432.10569	0.00004	Flavonoids

Table 3 Key target network attribute table

Name	Betweenness	Closeness	Degree
SLC7A11	0.066168	0.480315	6
GPX4	0.002891	0.472868	6
SQSTM1	0.049908	0.508333	12
MAP1 LC3B	0.058196	0.472868	9
BECN1	0.091432	0.512605	12
NFE2 L2	0.698875	0.72619	39

GPX4: Glutathione peroxidase 4.