Antibacterial potential of Euphorbia canariensis against Pseudomonas aeruginosa bacteria causing respiratory tract infections

Figures & data

Table 1. Phytochemical profiling of ECEE by LC-MS/MS analysis (positive mode ESI).

Metabolite Name	Adduct	RT. (min.)	Molecular Formula	Precursor m/z	Error	MS/MS	Class
Triethanolamine	[M + H]^+	1.201	C6H15NO3	150.1123	0.2	62, 68, 70, 86, 92, 108, 132, 150	1,2-aminoalcohols
Choline	[M]^+	1.214	C5H14NO	104.1053	9.5	56, 57, 58, 59, 60, 73, 86, 88, 103, 104	Cholines
Harmaline	[M + H]^+	1.215	C13H14N2O	215.1013	−3.7	59, 75, 89, 95, 109, 127, 144, 157, 172, 180., 187, 214, 215	Harmala alkaloids
Trigonelline	[M + H]^+	1.321	C7H7NO2	138.0544	0	51, 53, 65, 78, 84, 92, 93, 94, 123, 138,	Alkaloids and derivatives
Nicotine	[M + H]^+	8.227	C10H14N2	163.111	5.1	53, 57, 79, 91, 105, 115, 120, 128, 132, 135, 145, 163	Pyrrolidinylpyridines
4-Hydroxy-3-methoxy cinnamaldehyde	[M + H]^+	8.504	C10H10O3	179.0713	−2.2	55, 77, 79, 91, 109, 119, 136, 147, 179	Methoxyphenols
Kaempferol-3-O-alpha-L-rhamnoside	[M + H]^+	8.569	C21H20O10	433.115	1	71, 85, 103, 129, 147, 285, 287, 416, 433	Flavonoid-3-O-glycosides
3-(4-Hydroxy-3-methoxyphenyl)prop-2-enoicacid	[M + H]^+	8.615	C10H10O4	195.0977	8.2	55, 77,91,93,107, 121, 131,195	Hydroxycinnamic acids
Formononetin	[M + H]^+	11.108	C16H12O4	269.0216	0.1	116, 125,167, 191, 209, 269	4′-O-methylisoflavones
Scopoletin	[M + H]^+	11.818	C10H8O4	193.0848	3.7	51, 65, 77, 79, 89, 93, 102, 105, 137, 150, 165, 178, 193,	7-hydroxycoumarins
(-)-Riboflavin		12.285	C17H20N4O6	377.2052	6.3	119, 377	Flavins
Caffeine	[M + H]^+	13.112	C8H10N4O2	195.1018	−0.8	53, 77, 91, 109, 118, 150, 165, 195	Xanthines
Chlorogenic acid	[M + H]^+	13.61735	C16H18O9	355.1678	−1	77, 105, 115, 129, 145, 155, 195, 241, 355	Quinic acids and derivatives
4-Aminohippuric acid	[M + H]^+	13.997	C9H10N2O3	195.0644	−0.4	51, 77, 93, 105, 109, 115, 195	Hippuric acids
Daphnetin	[M + H]^+	14.327	C9H6O4	179.0697	0	77, 91, 107, 136, 149, 179	7,8-dihydroxycoumarins
1-O-b-D-gluco pyranosyl sinapate	[M + H]^+	14.327	C17H22O10	387.1822	−2.8	97, 139, 271, 387	Hydroxycinnamic acid glycosides
Genistein	[M + H]^+	15.019	C15H10O5	271.1494	14.4	97, 139, 271	Isoflavones
Daidzein	[M + H]^+	15.134	C15H10O4	255.0985	0	95, 255	Isoflavones
all-trans-Retinoic acid	[M + H]^+	16.247	C20H28O2	301.2157	0.5	67, 79, 105, 121, 135, 147, 175, 199, 283, 301	Retinoids
3,4-Dimethoxycinnamic acid	[M + H]^+	17.186	C11H12O4	209.0813	−0.2	65, 89, 105, 118, 159, 191, 209	Coumaric acids and derivatives
Acacetin-7-O-neohesperidoside	[M + H]^+	18.917	C28H32O14	593.2272	5.2	593	Flavonoid-7-O-glycosides
Syringaldehyde	[M + H]^+	20.918	C9H10O4	183.0988	5.9	51, 68, 98, 105, 168, 183	Methoxyphenols
Naringenin	[M + H]^+	21.653	C15H12O5	273.0948	−0.8	129, 245, 273	Flavanones
Leupeptin hemisulfate salt	[M + H]^+	26.477	C20H38N6O4	427.3559	−0.1	57, 133, 161, 207, 215, 301,409, 427	Dipeptides
3 3′ 4′ 5-Tetrahydroxy-7-methoxyflavone	[M + H]^+	26.938	C16H12O7	317.1158	−0.8	81, 177, 195, 282, 317	Flavonols

Table 2. Impact of ECEE on the biofilm formation of P. aeruginosa isolates.

Biofilm formation	Before	After
None	1	6
Weak	2	6
Moderate	9	4
Strong	7	3

Figure 1. Impact of ECEE on the gene expression of the biofilm-forming isolates.

Figure 2. The bacterial load in lung tissues of the experimental groups. The two asterisks represent a significant difference (p < 0.05). The abbreviation (NS) denotes a non-significant difference (p > 0.05).

Figure 3. Histological features of the lung tissues stained with H&E of A) Group I with normal-sized alveoli separated by fibrous septa (blue arrows), normal-sized bronchioles (black arrows), and normal-sized blood vessels (red arrow) (×100). B) Group II with dilated destructed bronchioles (black arrows), surrounded by marked alveolar fibrosis (blue arrows), marked inflammation (green arrows), and vascular congestion (red arrow) (×100). C) Group III with normal-sized bronchioles (black arrows), surrounded by normal-sized alveoli (blue arrows) with focal alveolar and vascular congestion (red arrows) (×100). D) Group IV with dilated bronchiole (black arrow), surrounded by normal-sized alveoli (blue arrows) with focal alveolar congestion (red arrow) (×100).

Figure 4. Masson’s trichrome staining of A) Group I with bronchioles and vessels surrounded by minimal blue stained collagen (red arrows) (×100). B) Group II with dilated bronchioles surrounded by marked fibrosis (red arrows) (×100). C) Group III with minimal fibrosis (red arrows) (×100). D) Group IV with mild fibrosis (red arrows) (×100).

Figure 5. Immunostained lung sections of A) Group I exhibited a negative expression of caspase-3 (0.12%) (×100). B) Group II exhibited a strong positive caspase-3 expression (87.24%) (×100). C) Group III exhibited a weak positive caspase-3 expression (17.14%) (×100). D) Group IV exhibited a weak positive caspase-3 expression (18.16%). E) Group I presented a negative expression of NF-kB (2.63%) (×100). F) Group II exhibited a strong positive NF-kB expression (82.17%) (×100). G) Group III exhibiting a weak positive NF-kB expression (16.25%) (×100). H) Group IV exhibited a weak positive NF-kB expression (18.06%) (×100).

Figure 6. Impact of ECEE on the levels of A) NO and B) MDA. The two asterisks represent a significant difference (p < 0.05). The abbreviation (NS) denotes a non-significant difference (p > 0.05).

Figure 7. Influence of ECEE on the gene expression of A) IL-6 and B) IL-10. The two asterisks represent a significant difference (p < 0.05). The abbreviation (NS) denotes a non-significant difference (p > 0.05).

Data availability statement

All data that support the finding displayed in our study are included in the manuscript. Any further data is available upon request.