In vivo evaluation and mechanism prediction of anti-diabetic foot ulcer based on component analysis of Ruyi Jinhuang powder

Table 1 Statistical analysis of wound healing rate at different times

Time	0 d (%)	3 d (%)	7 d (%)	14 d (%)
Control group	0	10.41 ± 0.24	17.43 ± 0.13	33.73 ± 0.28
Experimental group	0	32.78 ± 0.46	38.14 ± 0.28	53.11 ± 0.07
Homogeneity of variance	—	√	√	√
P value	—	< 0.05	< 0.05	< 0.05

Table 2 Gas chromatography–mass spectrometry full spectrum identification results

No.	RT (min)	Relative content (%)	Qualitative	CAS	Name
1	26.36	0.59	51.56	512-61-8	(-)-α-Santalene
2	27.07	0.26	25.81	87-44-5	Caryophyllene
3	29.71	1.86	65.77	495-60-3	α-Zingiberene
4	29.83	0.64	14.58	495-61-4	6-methyl-2-(4-methylcyclohex-3-enyl)hept-1,5-diene
5	30.64	1.95	60.85	20307-83-9	β-Sesquiphellandrene
6	30.7	1.33	80.1	644-30-4	α-Curcumene
7	34.21	0.38	26.47	56192-70-2	(Z)-a-Atlantone
8	34.27	0.23	61.92	1139-30-6	Caryophyllene oxide
9	34.36	0.30	19.52	83173-76-6	2-Methyl-1-(4-methylphenyl)-3-buten-1-ol
10	34.52	0.25	58.19	108549-47-9	(6Z)-2-Methyl-6-(4-methyl-3-cyclohexen-1-ylidene)-2-hepten-4-one
11	35.47	0.58	29.36	639-99-6	Elemol
12	35.78	0.28	58.71	145512-84-1	Trans-Sesquisabinene hydrate
13	36	0.37	62.02	58334-55-7	Zingiberenol
14	36.57	0.74	86.3	6989-21-5	Atractylon
15	36.83	0.25	12.5	82508-14-3	2-methyl-6-(4-methylidenecyclohex-2-en-1-yl)hept-2-en-4-one
16	37.23	0.77	22.65	1209-71-8	γ-Eudesmol
17	37.41	20.74	91.81	180315-67-7	Tumerone
18	37.67	13.80	22.04	1460-73-7	Agarospirol
19	37.81	1.75	38	112-39-0	Methyl palmitate
20	38.41	1.26	41.2	473-16-5	a-Eudesmol
21	38.48	0.72	9.97	515-20-8	Costol
22	38.65	9.06	76.48	473-15-4	β-Eudesmol
23	39.1	24.33	97.75	532-65-0	ar-Turmerone
24	39.28	0.24	11.67	19888-00-7	4,8-Cycloundecadien-1-ol, 6,6,9-trimethyl-2-methylene-, (1R,4E,8E)-
25	39.47	0.23	4.42	65646-68-6	N-(4-hydroxyphenyl)retinamide
26	39.64	0.39	8.22	19912-67-5	(+)-Epicubenol
27	39.9	0.85	49.47	30666-87-6	2-Methyl-6-(4-methylphenyl)-4-heptanone
28	40.16	1.05	49.07	72441-71-5	(6R,7R)-Bisabolone
29	40.53	2.23	89.2	108645-54-1	(E)-Atlantone
30	40.89	0.65	82.79	120681-80-3	2-Cyclohexene-1-propanol,g-methyl-4-methylene-a-(2-methyl-1-propen-1-yl)
31	41.58	0.45	75.15	112-61-8	Octadecanoic acid, methyl ester
32	41.88	1.87	16.03	112-62-9	Methyl oleate
33	41.98	0.65	7.88	1937-62-8	trans-octadec-9-enoicacidmethylester
34	42.55	1.03	26.03	112-63-0	methyl linoleate
35	43.54	0.42	42.85	301-00-8	9,12,15-Octadecatrienoic acid, methyl ester
36	43.61	0.22	4.63	29550-55-8	Teresantalol(7CI)
37	44.58	0.24	88.5	69301-27-5	2-(1,5-Dimethyl-4-Hexenyl)-5-Methyl-Phenol
38	43.85	0.84	6.23	38142-57-3	2-Methyl-6-(p-tolyl)hept-2-en-4-ol
39	45.01	1.11	28.14	3218-36-8	4-Biphenylcarboxaldehyde
40	46.86	0.84	75.64	4666-84-6	Proximadiol
41	47.75	0.46	98.07	949081-10-1	(S)-3-Methyl-6-((S)-6-methyl-4-oxohept-5-en-2-yl)cyclohex-2-enone
42	48.71	0.24	9.54	2566-90-7	Methyl 4,7,10,13,16,19-cis-docosahexanenoate
43	51.08	2.31%	83.12	1957-10-3	Palmitic acid

Table 3 Ultra-performance liquid chromatography full spectrum identification results

No.	Name	CAS	Molecular weight	RT (min)	Relative concentration (μg/mL)
1	DL-Arginine	7200-25-1	174.11175	0.783	24.44253696
2	Nitrosobis(2-oxopropyl)amine	60599-38-4	158.06914	0.817	15.74503993
3	Gluconic acid	526-95-4	196.05765	0.821	82.48616363
4	D-(+)-Proline	344-25-2	115.06357	0.846	15.35704499
5	Cabotegravir	1051375-10-0	405.11182	0.847	20.08010135
6	α,α-Trehalose	99-20-7	342.11623	0.847	409.0449144
7	D-(-)-Quinic acid	77-95-2	192.06275	0.85	56.2670819
8	Isocitric acid	320-77-4	192.02637	0.939	63.29259087
9	Citric acid	77-92-9	192.02637	1.169	145.2856943
10	Gallic acid	149-91-7	170.02078	1.899	23.57871448
11	Chlorogenic acid	327-97-9	354.09527	5.332	44.58986172
12	Catechin	88191-48-4	290.07917	5.431	98.07463825
13	methyl chlorogenate	123483-19-2	368.11081	5.501	18.15074983
14	6-Acetylcodeine	6703-27-1	341.16237	5.886	12.3548527
15	2-(3,4-Dihydroxyphenyl)ethyl 3-O-(6-deoxy-β-L-mannopyranosyl)-6-O-[(2E)-3-(3,4-dihydroxyphenyl)-2-propenoyl]-β-D-glucopyranoside	61303-13-7	624.20617	6.092	20.78761602
16	(3R,5R)-1,3,5-Trihydroxy-4-{[(2E)-3-(4-hydroxy-3-methoxyphenyl)-2-propenoyl]oxy}cyclohexanecarboxylic acid	2613-86-7	368.11084	6.196	201.3429867
17	Emodin-8-Beta-D-Glucoside	23313-21-5	432.10571	6.579	7.280885459
18	2-[4-[3-[3,4-dihydroxy-4-(hydroxymethyl)oxolan-2-yl]oxy-4,5-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxyphenyl]-7-hydroxy-2,3-dihydrochromen-4-one	74639-14-8	550.16902	6.625	13.98565399
19	Baicalin	21967-41-9	446.0852	6.663	16.85569046
20	Liquiritin	551-15-5	418.12676	6.737	22.91985259
21	Naringin	10236-47-2	580.17964	6.959	26.36327029
22	Hesperidin	520-26-3	610.19038	7.191	85.82076776
23	Azelaic acid	123-99-9	188.1042	7.548	4.773123466
24	Berberine	2086-83-1	335.11525	8.242	232.2489146
25	isosakuranetin-7-O-rutinoside	14259-47-3	594.19556	8.493	4.34803523
26	Tinnevellin glucoside	80358-06-1	408.14228	8.786	13.59498227
27	Daidzein	486-66-8	254.05778	9.009	13.05981793
28	Chrysophanol 8-O-β-D-glucoside	13241-28-6	416.11099	9.01	17.88080565
29	Licoricesaponin G2	118441-84-2	838.39998	9.824	7.195669531
30	Luteolin	491-70-3	286.04776	9.909	1.034661042
31	(15Z)-9,12,13-Trihydroxy-15-octadecenoic acid	95341-44-9	330.24071	9.938	19.41925634
32	Rheic acid	478-43-3	284.03211	11.029	81.8044913
33	Bisdemethoxycurcumin	24939-16-0	308.10496	11.328	153.9247824
34	6-Hydroxy-2-naphthoic acid	16712-64-4	188.04652	11.328	7.047135466
35	Demethoxycurcumin	22608-11-3	338.11567	11.473	151.9906609
36	5,7-dihydroxy-2-(4-hydroxy-3-methoxyphenyl)-6-(3-methylbut-2-enyl)-2,3-dihydrochromen-4-one	76735-58-5	370.14168	11.541	55.08087646
37	Curcumin	458-37-7	368.12606	11.619	307.1972754
38	Isoimperatorin	482-45-1	270.08918	11.847	1.025466018
39	Genistein	446-72-0	270.05279	12.256	153.8276373
40	(+/-)9-HODE	98524-19-7	296.23511	13.146	23.43829567
41	(+)-ar-Turmerone	532-65-0	216.15135	13.605	23.84749189
42	Indane	496-11-7	118.07843	13.605	27.63249372
43	Prespatane	100387-71-8	204.18777	13.956	2.010509748
44	(+)-Nootkatone	4674-50-4	218.16696	14.497	16.6606139
45	Aristolone	6831-17-0	218.16696	14.588	45.88543598
46	2,2'-Methylenebis(4-methyl-6-tert-butylphenol)	119-47-1	340.24015	16.788	25.70628216

RT: Radiotherapy.

Table 4 Ruyi Jinhuang powder “Chinese medicine-ingredients-target” correspondence

CAS	Name	GenecardID	Attribution
512-61-8	(-)-α-Santalene	PTGS2, CHRM2	JH
87-44-5	Caryophyllene	PTGS2, CHRM2	BZ, JH, CZ, ZHP, TNX
495-60-3	α-Zingiberene	PTGS2, DPP4	JH
20307-83-9	β-Sesquiphellandrene	PTGS2	JH
644-30-4	α-Curcumene	PTGS2, TOP2A, DPP4	JH
1139-30-6	Caryophyllene oxide	CHRM2, DPP4	ZHP, TNX
639-99-6	Elemol	CHRM2	ZHP, JH
58334-55-7	Zingiberenol	CHRM2	JH
6989-21-5	Atractylon	SCN5A, CHRM4, HTR2A, CHRM2, OPRM1	CZ
82508-14-3	2-methyl-6-(4-methylidenecyclohex-2-en-1-yl)hept-2-en-4-one	PTGS2, CHRM2	JH, ZHP, CZ
112-39-0	Methyl palmitate	PTGS2, IL-10, TNF, IL-6	JH, ZHP, BZ, HB, THF
473-15-4	beta-Eudesmol	CHRM2	ZHP, JH, CZ
532-65-0	(6S)-2-methyl-6-(4-methylphenyl)hept-2-en-4-one	DPP4, PTGS2, CHRM2x, ampC	JH
112-62-9	Methyl oleate	RXRA	ZHP, THF
1937-62-8	trans-octadec-9-enoicacidmethylester	RXRA	ZHP, THF
112-63-0	methyl linoleate	PTGS2, RXRA	BZ, THF, ZHP, TNX
301-00-8	9,12,15-Octadecatrienoic acid, methyl ester, (Z,Z,Z)-	PTGS2, RXRA	THF
38142-57-3	2-Methyl-6-(p-tolyl)hept-2-en-4-ol	PTGS2, CHRM2, DPP4, ampC, RXRA	THF, ZHP, TNX
1957—10—3	Palmitic acid	PTGS2, IL-10, TNF, PCYT1A	DH, ZHP, THF
149-91-7	Gallic acid	PTGS2, PTPN1, TOP2A	DH, TNX, CP, GC, JH
23313-21-5	Emodin-8-Beta-D-Glucoside	TOP2A	DH
458-37-7	Curcumin	PTGS2, PTPN1	JH
22608-11-3	Demethoxycurcumin	PTGS2, PTPN1, AKR1B1, ABCB1, SCN5A	JH
520-26-3	Hesperidin	PTGS2	CP
14259-47-3	isosakuranetin-7-O-rutinoside	TOP2A	CP
10236-47-2	Naringin	TOP2A, CDKN1A, TNF, RASGRF2, RAF, PTGS2, ampC, MTTP, APOB, mvaA, PPARA	CP, GC
21967-41-9	Baicalin	PTPN1, TOP2A	DH
88191-48-4	Catechin	CHRM2	DH, CP
491-70-3	Luteolin	PTGS2, DPP4, EGFR, IL10, CDK4, TNF, IL6, NFKBIA, psdA1, IL2, TOP2A, SLC2A4, INSR, MET	CP, ZHP
551-15-5	Liquiritin	PTGS2	GC
478-43-3	Rheic acid	PTGS2, AKR1B1	DH
2086-83-1	Berberine	PTGS2, RXRA	HB
77-92-9	Citric acid	PTGS2, AKR1B1, GRIN2A, GRIA2, PTPN1	DH
320-77-4	Isocitric acid	PTGS2	DH
7200-25-1	DL-Arginine	PTGS2	THF
446-72-0	Genistein	PTGS2, EGFR, TNF, SELE, IL8	GC
482-45-1	Isoimperatorin	PTGS2	BZ
486-66-8	Daidzein	PTGS2, RXRA	GC
6831-17-0	Aristolone	CHRM2, PTGS2	BZ
532-65-0	(+) -ar-Turmerone	PTGS2, RXRA, CHRM2, ampC, DPP4	JH
327-97-9	Chlorogenic acid	PTGS2	HB
77-95-2	D-(-)-Quinic acid	PTGS2	HB
1460-73-7	Agarospirol	CHRM2	ZHP

TNF: Tumor necrosis factor; IL: Interleukin; PTGS2: Posttranscriptional silencing; TOP2A: Topoisomerase II alpha; CHRM2: Muscarinic acetylcholine receptor M2; RXRA: Retinoic X receptor alpha; DPP4: Dipeptidyl peptidase 4; SCN5A: Sodium voltage-gated channel alpha subunit 5; HTR2A: 5-hydroxytryptamine (serotonin) receptor 2A; OPRM1: Opioid Receptor Mu 1; EGFR: Epidermal Growth Factor Recepto; PTPN1: Protein tyrosine phosphatase nonreceptor type 1; THF: Trichosanthin (Tian Hua); DH: Rhubarb (Da Huang); HB: Phellodendron (Huang Bai); JH: Turmeric (Jiang Huang); BZ: Angelica dahurica (Bai Zhi); TNX: Arisaema (Tian Nan Xing); CZ: Atractylodes lancea (Cang Zhu); HP: Magnolia officinalis (Hou Po); CP: Pericarpium Citri Reticulatae (Chen Pi); GC: Glycyrrhiza uralensis (Gan Cao).

Table 5 Docking parameters of chemical components and target protein molecules

Name	Target	LibDockScore
Luteolin	TOP2A	104.333
Methyl palmitate	PTGS2	114.221
ar-Turmeric	PTGS2	91.0625
	CHRM2	88.3765
Methyl Linoleate	RXRA	138.583
	PTGS2	130.838
Palmitic acid	PTGS2	114.349
Demethoxycurcumin	PTGS2	135.479
Naringin	TOP2A	141.197
	PTGS2	164.305

PTGS2: Posttranscriptional silencing; TOP2A: Topoisomerase II alpha; CHRM2: Muscarinic acetylcholine receptor M2; RXRA: Retinoic X receptor alpha.