Metabolomics coupled with network pharmacology study on the protective effect of Keguan-1 granules in LPS-induced acute lung injury

Figures & data

Figure 1. Effects of KG-1 on TNF-α (A), IL-6 (B), CXCL1 (C), CXCL2 (D), and lung histopathological changes (E) in ALI mice. Blue, red, green, violet columns represent the control, model, KG-1, and DXMS group, respectively. **p < 0.01 versus the control group, ^#p < 0.05, ^##p < 0.01 versus the model group. Data are presented as the mean ± SEM (n = 8).

Figure 2. Metabolome profiling for KG-1 anti- ALI in ESI + model. (A,B) PCA and OPLS-DA plots among the control, model and KG-1 groups. (C,E) OPLS-DA plots in the control group [R2X (cum) = 0.214, R2Y (cum) = 0.997, Q2 (cum) = 0.759] or KG-1 group [R2X (cum) = 0.545, R2Y (cum) = 1, Q2 (cum) = 0.869] in comparison to the model group. (D,F) S-plot analysis in the control group or KG-1 group in comparison to the model group. (G) Venn diagram, the intersection of two groups means was the set of variables related to KG-1 anti-ALI. QC sample (n = 5, yellow circle), the control group (n = 8, green box), the model group (n = 8, blue triangle), the KG-1 group (n = 8, red diamond).

Figure 3. Metabolome profiling for KG-1 anti- ALI in ESI- model. (A,B) PCA and OPLS-DA plots among the control, model and KG-1 groups. (C,E) OPLS-DA plots in the control group [R2X (cum) = 0.38, R2Y (cum) = 0.986, Q2 (cum) = 0.881] or KG-1 group [R2X (cum) = 0.348, R2Y (cum) = 0.996, Q2 (cum) = 0.926]. (D,F) S-plot analysis in the control group or KG-1 group in comparison to the model group. (G) Venn diagram, the intersection of two groups means was the set of variables related to KG-1 anti-ALI. QC sample (n = 5, yellow circle), control group (n = 8, green box), model group (n = 8, blue triangle), KG-1 group (n = 8, red diamond).

Figure 4. Global metabolome outline. (A) Radar chart of 12 metabolites with relative abundance among the control group (blue line), model group (red line) and KG-1 group (green line). (B) Pathway enrichment of metabolites. (C) Network schematic diagram of metabolites and involved pathways. In the KG-1 vs model group, the metabolites in the pink box were increased in the KG-1 group; the metabolites in the blue box were decreased in the KG-1 group; the metabolites without box were not detected. Data are presented as the mean ± SEM (n = 8).

Table 1. The detailed information about 12 metabolites among the control, model and KG-1 groups.

Model	MZ	RT/min	Name	Formula	Control group/Model group				KG-1 group/Model group
					FC	P(corr)	VIP	p Value	FC	P(corr)	VIP	p Value
ESI+	784.6833	21.38	Glucosylceramide (d18:1/22:0)	C46H89NO8	0.327	0.727	1.825	0.001	0.111	−0.954	1.701	0.000
ESI+	799.6883	18.48	SM(d17:1/24:1(15Z))	C46H92N2O6P	0.139	0.764	1.924	0.000	0.159	−0.760	1.356	0.000
ESI+	301.2248	16.35	Retinoic acid	C20H28O2	0.247	0.770	1.933	0.000	0.373	−0.781	1.389	0.000
ESI+	792.6038	16.11	PC(22:5(7Z,10Z,13Z,16Z,19Z)/P-16:0)	C46H83NO7P	0.322	0.641	1.628	0.006	0.362	−0.614	1.217	0.011
ESI-	881.5313	20.47	PI(22:6(4Z,7Z,10Z,13Z,16Z,19Z)/16:0)	C47H79O13P	0.149	0.724	1.001	0.001	0.123	−0.737	1.183	0.001
ESI-	736.4756	16.16	PE(22:4(7Z,10Z,13Z,16Z)/14:1(9Z))	C41H72NO8P	0.245	0.841	1.103	0.000	0.383	−0.606	1.053	0.012
ESI+	345.1993	20.80	11-Dehydrocorticosterone	C21H28O4	0.180	0.608	1.667	0.011	0.207	−0.601	1.211	0.013
ESI+	265.1125	7.13	Thiamine	C12H17N4OS	4.018	−0.593	1.549	0.012	6.506	0.751	1.343	0.000
ESI+	139.0548	3.43	Urocanate	C6H6N2O2	0.348	0.669	1.807	0.005	0.262	−0.771	1.452	0.000
ESI+	175.0906	2.06	(Indol-3-yl)acetamide	C10H10N2O	0.262	0.567	1.443	0.021	0.300	−0.549	1.043	0.027
ESI+	110.0638	1.57	2-Aminophenol	C6H7NO	0.324	0.634	1.655	0.010	0.373	−0.567	1.077	0.021
ESI+	180.1063	17.94	l-Homophenylalanine	C10H13NO2	4.333	−0.537	1.355	0.038	7.578	0.645	1.175	0.006

Figure 5. The “compound-target-potential metabolite” interaction network of KG-1 in the treatment of ALI. (A) Overall interaction outline. (B) For those directly involved metabolites and targets in KG-1. The red triangles represent the putative compounds of KG-1; the sky-blue dots represent KG-1 related to putative targets; the purple dots represent potential metabolites related to putative targets.

Figure 6. The “compound-target-potential metabolite” simplified interactive network. The navy blue, red, pink, purple, light-green, dark-green, light-yellow triangles represent the putative active compounds of ZBM (Zhebeimu), LQ (Lianqiao), KXR (Kuxingren), SY (Sangye), JLSJ (Jinyinhua-Liaoqiao-Sangye-Juhua), JS (Jinyinhua-Sangye), JSYK (Jinyinhua-Sangye-Yiyiren-Kuxingren), respectively; the yellow dots represent the candidate metabolites; the sky-blue dots represent the shared targets of KG-1 and potential metabolites.