Betaine combined with traditional Chinese medicine ointment to treat skin wounds in microbially infected diabetic mice

Table 1 Compound list

Compound	CAS
Deoxyshikonin	43043-74-9
Shikonin	517-89-5
Lithospermoside	63492-69-3
β,β-Dimethylacrylshikonin	24502-79-2
β,β-Dimethylacrylalkannin	34539-65-6
Lithospermic acid	28831-65-4
β-Acetoxyisovalerylshikonin	69091-17-4
Isobutylshikonin	52438-12-7
Acetylshikonin	24502-78-1
Isobavachalcone	20784-50-3
Bavachin	19879-32-4
Isobavachin	31524-62-6
Bavachinin	19879-30-2
4'-O-Methylbavachalcone	20784-60-5
Psoralen	66-97-7
Corylin	53947-92-5
Psoralidin	18642-23-4
Astragalin	480-10-4
Methoxsalen	298-81-7
Bakuchiol	10309-37-2
Angelicin	523-50-2
Neobavaisoflavone	41060-15-5
Cinnamaldehyde	104-55-2
Cinnamic acid	621-82-9
Cinnamon alcohol	104-54-1
Cinnamyl acetate	103-54-8
Trans-cinnamic acid	140-10-3
p-Hydroxy-cinnamic acid	7400-08-0
2-Methoxycinnamic acid	6099-03-2
Methyl 4-hydroxycinnamate	3943-97-3
3,4,5-Trimethoxycinnamic acid	90-50-6

Table 2 The minimum inhibitory concentrations of gromwell root compounds (μg/mL)

Compound	Klebsiella pneumoniae	Pseudomonas aeruginosa	Acinetobacter baumannii	Staphylococcus aureus	Candida albicans	Cryptococcus neoformans	Helicobacter pylori
Shiconin	> 128	> 128	> 128	> 128	> 128	> 128	2
Lithospermoside	> 128	> 128	> 128	> 128	8	4	> 128
β,β-Dimethylacrylshikonin	> 128	> 128	> 128	> 128	> 128	> 128	32
β,β-Dimethylacrylalkannin	> 128	> 128	> 128	> 128	32	8	32
Lithospermic acid	> 128	> 128	> 128	> 128	> 128	> 128	> 128
β-Acetoxyisovalerylshikonin	> 128	> 128	> 128	> 128	> 128	> 128	8
Isobutylshikonin	> 128	> 128	> 128	> 128	> 128	> 128	8
Acetylshikonin	> 128	> 128	> 128	> 128	> 128	8	4
Deoxyshikonin	> 128	> 128	> 128	> 128	> 128	8	2

Table 3 The minimum inhibitory concentrations of psoralen compounds (μg/mL)

Compound	Klebsiella pneumoniae	Pseudomonas aeruginosa	Acinetobacter baumannii	Staphylococcus aureus	Cryptococcus neoformans	Candida albicans	Helicobacter pylori
Isobavachalcone	> 128	> 128	> 128	8	1	8	8
Bavachin	> 128	> 128	> 128	> 128	> 128	> 128	8
Bavachinin	> 128	> 128	> 128	16	32	> 128	16
4'-O-Methylbavachalcone	> 128	> 128	> 128	> 128	> 128	> 128	8
Neobavaisoflavone	> 128	> 128	> 128	64	32	> 128	16
Isobavachin	> 128	> 128	> 128	16	32	> 128	> 128
Bakuchiol	> 128	> 128	> 128	32	16	> 128	> 128
Methoxsalen	> 128	> 128	> 128	64	> 128	> 128	> 128
Angelicin	> 128	> 128	> 128	> 128	> 128	> 128	> 128
Psoralen	> 128	> 128	> 128	128	> 128	> 128	> 128
Corylin	> 128	> 128	> 128	> 128	> 128	> 128	> 128
Psoralidin	> 128	> 128	> 128	> 128	> 128	> 128	> 128
Astragalin	> 128	> 128	> 128	> 128	> 128	> 128	> 128

Table 4 The minimum inhibitory concentrations of cinnamon compounds (μg/mL)

Compound	Staphylococcus aureus	Pseudomonas aeruginosa	Klebsiella pneumoniae	Acinetobacter baumannii	Candida albicans	Cryptococcus neoformans	Helicobacter pylori
Cinnamaldehyde	64	> 128	> 128	> 128	128	64	8
Cinnamic acid	> 128	> 128	> 128	> 128	> 128	128	128
Cinnamon alcohol	> 128	> 128	> 128	> 128	> 128	> 128	128
Cinnamyl acetate	> 128	> 128	> 128	> 128	> 128	> 128	> 128
Trans-cinnamic acid	> 128	> 128	> 128	> 128	128	64	128
p-Hydroxy-cinnamic acid	> 128	> 128	> 128	> 128	> 128	> 128	128
2-Methoxycinnamic acid	> 128	> 128	> 128	> 128	> 128	128	> 128
Methyl 4-hydroxycinnamate	> 128	> 128	> 128	> 128	> 128	128	> 128
Ethyl 4-methoxycinnamate	> 128	> 128	> 128	> 128	> 128	128	16
3,4,5-Trimethoxycinnamic acid	> 128	> 128	> 128	> 128	> 128	> 128	> 128

Table 5 Antibacterial spectrum of betaine (μg/mL)

Strain	MIC (μg/mL)
Bacillus subtilis	4
Newman	4
USA300	8
Candida albicans	16
Cryptococcus neoformans	16
Candida tropicalis	32
Escherichia coli	> 1024
Staphylococcus haemolyticus	> 1024
Pseudomonas aeruginosa	> 1024
Proteus mirabilis	> 1024
Acinetobacter baumannii	> 1024
Klebsiella pneumoniae	> 1024
Morganella morganii subsp. morganii	> 1024
Enterobacter hormaechei	> 1024

MIC: Minimum inhibitory concentration.

Table 6 The minimum inhibitory concentrations of betaine ointment (μg/mL)

Strain	MIC (μg/mL)
	BO	Zicao
Newman	16	> 2048
USA300	32	> 2048
Candida albicans	64	> 2048
Cryptococcus neoformans	64	> 2048
Candida tropicalis	64	> 2048
Staphylococcus haemolyticus	> 512	> 2048
Pseudomonas aeruginosa	> 512	> 2048
Proteus mirabilis	> 512	> 2048
Acinetobacter baumannii	> 512	> 2048
Klebsiella pneumoniae	> 512	> 2048

MIC: Minimum inhibitory concentration; BO: Betaine ointment.

Table 7 Effect of betaine ointment on scab removal in skin wounds infected with Staphylococcus aureus

Strain	Group	Number of mice shed scabs (pcs)
		Day 10	Day 11	Day 12	Day 13	Day 14	Day 1	Total	Rate (%)
Newman	Mupirocin	0	2	0	0	1	155	5	83.3a
	Zicao	0	0	0	0	1	1	2	33.3a
	BO	0	1	0	0	2	0	3	50.0a
USA300	Mupirocin	2	0	0	2	0	0	4	66.7b
	Zicao	0	0	0	0	0	0	0	0.0b
	BO	2	0	0	1	0	2	5	83.3b

^aP < 0.0001.

^bP < 0.0001.

BO: Betaine ointment.

Table 8 Effects of betaine ointment on scab removal from skin wounds in diabetic mice infected with Staphylococcus aureus

Group	Number of mice shed scabs (pcs)
	Day 10	Day 11	Day 12	Day 13	Day 14	Day 15	Total	Rate (%)
PBS	0	0	0	0	0	0	0	0.0
Mupirocin	1	1	2	0	0	0	4	80.0c
BO	0	1	1	1	1	0	4	80.0c

^cP < 0.05 vs phosphate buffer solution.

PBS: Phosphate buffer solution; BO: Betaine ointment.