BanXiaXieXin decoction treating gastritis mice with drug-resistant Helicobacter pylori and its mechanism

Table 1 Chromatographic conditions

Time (min)	Parameter
0	A: 0%, B: 100%
10	A: 50%, B: 50%
13	A: 95%, B: 5%
14	A: 0%, B: 100%
15	A: 0%, B: 100%

Table 2 Primer information

No.	Primer	Sequence (5’ to 3’)	Company
1	UREA F	GCCAATGGTAAATTAGTT	Shanghai Invitrogen Biotech Co., Lt
2	UREA R	CTCCTTAATTGTTTTTAC
3	UREB F	TCTATCCCTACCCCACAACC
4	UREB R	CCATCCACGAACACATGGTA
5	CagA F	ACCCCTAGTCGGTAATG
6	CagA R	GCTTTAGCTTCTGATACTGC
7	VacA F	GTCAGCATCACACCGCAAC
8	VacA R	CTGCTTGAATGCGCCAAAC
9	16sRNA F	CTGGAGAGACTAAGCCCTCC
10	16sRNA R	AGGATCAAGGTTTAAGGATT

Table 3 Antibody information

Name	Art. No.	Company
CagA (A-10)	sc-32746	Santa Cruz
VacA	sc-28368	Santa Cruz
m-IgGk BP-HRP	sc-516102	Santa Cruz
GAPDH Ab	AF7021	Affinity Biosciences
Goat anti-rabbit IgG (H+L) HPR	S0001	Affinity Biosciences

Table 4 Minimum inhibitory concentration of Banxia Xiexin decoction aqueous extract against Helicobacter pylori (μg/mL)

Strain	Drug-resistanct strain	BXXXT aqueous extract	Berberine
26695	Sensitive	512	1024
G27	Sensitive	512	1024
NSH57	Sensitive	256	512
BHKS159	Resistant to levofloxacin, clarithromycin and metronidazole	512	1024
HPBS001	Resistant to levofloxacin, clarithromycin and metronidazole	512	1024
HPBS002	Resistant to metronidazole	512	1024
HPBS003	Resistant to clarithromycin	512	1024
HPBS004	Resistant to levofloxacin	512	1024
HPBS005	Resistant to levofloxacin and metronidazole	256	1024
HPBS006	Resistant to clarithromycin and metronidazole	256	1024
HPBS007	Resistant to clarithromycin	512	1024
HPBS010	Resistant to metronidazole, clarithromycin and levofloxacin	512	2048
HPBS011	Resistant to metronidazole and clarithromycin	512	1024
HPBS013	Resistant to metronidazole, clarithromycin and levofloxacin	512	1024
HPBS014	Resistant to metronidazole, clarithromycin, amoxicillin and levofloxacin	512	1024

BXXXT: Banxia Xiexin decoction. The minimum inhibitory concentrations of the drugs for sensitive and drug-resistant strains are amoxicillin ≥ 0.5 μg/mL, clarithromycin ≥ 1.0 μg/mL, levofloxacin ≥ 2.0 μg/mL, and metronidazole ≥ 8.0 μg/mL.

Table 5 Information of main pharmacodynamic components of Banxia Xiexin decoction aqueous extract

Name	Molecular structural formula
Berberine	COC1=C(OC)C2=C[N+]3=C(C=C2C=C1)C1=CC2=C(OCO2)C=C1CC3
Baicalin	C1=CC=C(C=C1)C2=CC(=O)C3=C(C(=C(C=C3O2)O[C@H]4[C@@H]([C@H]([C@@H]([C@H](O4)C(=O)O)O)O)O)O)O
Luteolin	OC1=CC(O)=C2C(=O)C=C(OC2=C1)C1=CC(O)=C(O)C=C1
Gallic acid	OC(=O)C1=CC(O)=C(O)C(O)=C1
Gingerol	CCCCCC(O)CC(=O)CCC1=CC(OC)=C(O)C=C1
Wogonoside	COC1=C(O)C=C(O)C2=C1OC(=CC2=O)C1=CC=CC=C1
Rosmarinic acid	OC(=O)[C@H](CC1=CC(O)=C(O)C=C1)OC(=O)\C=C\C1=CC(O)=C(O)C=C1
Aloe-emodin	OCC1=CC2=C(C(O)=C1)C(=O)C1=C(C=CC=C1O)C2=O
Catechin	CCCCCC(CC(=O)CCC1=CC(=C(C=C1)O)OC)O
Naringenin	OC1=CC=C(C=C1)[C@@H]1CC(=O)C2=C(O)C=C(O)C=C2O1
Quercetin	OC1=CC(O)=C2C(OC(=C(O)C2=O)C2=CC(O)=C(O)C=C2)=C1

Table 6 Minimum inhibitory concentrations of main antibacterial components in Banxia Xiexin decoction aqueous extract (μg/mL)

Strain	MIC of drugs used alone		MIC of drugs used in combinations		FIC	Effect
	Berberine	Emodin	Berberine	Emodin
G27	1024	512	256	256	0.75	Addictive
26695	1024	512	256	128	0.50	Synergistic
BHKS159	1024	512	256	128	0.50	Synergistic
	Berberine	Luteolin	Berberine	Berberine
G27	1024	1024	512	512	1.00	Addictive
26695	1024	1024	512	256	0.75	Addictive
BHKS159	1024	1024	512	512	1.00	Addictive
	Luteolin	Gallic acid	Luteolin	Gallic acid
G27	1024	1024	256	256	0.50	Synergistic
26695	1024	1024	256	256	0.50	Synergistic
BHKS159	1024	1024	256	256	0.50	Synergistic
	Luteolin	Rosmarinic acid	Luteolin	Rosmarinic acid
G27	1024	1024	512	256	0.75	Addictive
26695	1024	1024	512	512	1.00	Addictive
BHKS159	1024	1024	512	256	0.75	Addictive
	Catechuic acid	Quercetin	Catechuic acid	Quercetin
G27	1024	1024	512	256	0.75	Addictive
26695	1024	1024	512	256	0.75	Addictive
BHKS159	1024	1024	512	256	0.75	Addictive
	Catechuic acid	Emodin	Catechuic acid	Emodin
G27	1024	512	512	256	1.00	Addictive
26695	1024	512	512	256	1.00	Addictive
BHKS159	1024	512	256	256	0.75	Addictive

MIC: Minimum inhibitory concentration.

Table 7 Information of related proteins identified by transcriptome analysis after Banxia Xiexin decoction treatment

Uniprot Accession Number	Protein name	Protein amino acid sequence	Pvalue	Reliability
E6NPU2	Urease subunit alpha OS = Helicobacter pylori (strain F57), OX = 866346, GN = urea, PE = 3, SV = 1	MKLTPKELDKLMLHYAGELARKRKEKGIKLNYVEAVALISAHIMEEARAGKKSAAELMQEGRTLLKPDDVMDGVASMIHEVGIEAMFPDGTKLVTVHTPIESNGKLVPGELFLKNEDITINEGKKAVSVKVKNVGDRPVQIGSHFHFFEVNRCLDFDREKTFGKRLDIASGTAVRFEPGEEKSVELIDIGGNRRIFGFNALVDRQADNESKKIALHRAKERGFHGAKSDDNYVKTIKE	0.001211988	High
A0A2A6SFH9	Urease (fragment) OS = Helicobacter pylori, OX = 210, GN = BB479_08100, PE = 4, SV = 1	MKLTPKELDKLMLHYAGELARKRKEKGIKLNYVEAVALIXAHIMEEARAGKKTAAELMQEGRTLLKPDDVMDGVASMIHEVGIEAMFPDGTKLVTVHTXIEANGKLVPGELFLKNEDITINEGKKAVSVKVKNVGDRPVQIGSH	0.033815784	High
A0A0L0QH58	Urease subunit alpha OS = Helicobacter pylori, OX = 210, GN = urea, PE = 3, SV = 1	MKLTPKELDKLMLHYAGELAKKRKEKGIKLNYVEAVALISAHIMEEARAGKKSAAELMQEGRTLLKPDDVMDGVASMIHEVGIEAMFPDGTKLVTVHTPIEANGKLVPGELFLKNEDITINEGKKAVSVKVKNVGDRPVQIGSHFHFFEVNRCLDFDREKTFGKRLDIASGTAVRFEPGEEKSVELIDIGGNRRIFGFNALVDRQADNESKKIALHRAKERGFHGAKSDDNYVKTIKE	0.033744196	High
N4TND2	Urease accessory protein UreG OS = Helicobacter pylori Hp A-11, OX = 992035, GN = ureG, PE = 3, SV = 1	MVKIGVCGPVGSGKTALIEALTRHMSKDYDMAVITNDIYTKEDAEFMCKNSVMPRERIIGVETGGCPHTAIREDASMNLEAVEEMHGRFPNLELLLIESGGDNLSATFNPELADFTIFVIDVAEGDKIPRKGGPGITRSDLLVINKIDLAPYVGADLKVMERDSKKMRGEKPFIFTNIRAKEGLNDVIAWIKRNALLED	0.016839824	High
Q8RRP6	Cytotoxin associated protein CagA (Fragment), OS = Helicobacter pylori, OX = 210, GN = cagA, PE = 4, SV = 1	ALADLKNFSKEQLAQQAQKNESFNAGKKFEFSQSVRNGVNGTLVGNGFSQAEATTLSKNFSDIKKELNAKLGNFNNNNINGLKNSTEPIYAKVNKKETGQAASPEEPIYTQVAKKVNAKIDRLNQIASGLGVVGQAAGFPLKRHDKVDDLSKVGRSVSPEPIYATIDDLGGPFPLKRHDKVDNLSKVGRSVSPEPIYATIDDLGGPFPLKRHDKVDNLSKVGLSRNQELTQKIDNLSQAVSEAKAGFFGNLEQTIDKLKDSTKHNVVNLWAESAKKVPASLSAKLDNYA	0.03652953	High