Therapeutic mechanism of Arctium lappa extract for post-viral pneumonia pulmonary fibrosis: a metabolomics, network pharmacology analysis and experimental verification

doi:10.12122/j.issn.1673-4254.2025.06.08

Abstract

Abstract:

Objective To explore the therapeutic mechanism of Arctium lappa extract for treatment of Post-Viral Pneumonia Pulmonary Fibrosis (PPF). Methods The chemical constituents of Arctium lappa extracts were identified using UHPLC-Q-TOF-MS/MS. Mouse models of pulmonary fibrosis established by tracheal instillation of bleomycin were treated with Arctium lappa extract, and body weight changes were recorded and lung tissue pathology was examined using HE and Masson staining. Metabolomics analysis was used to identify the differential metabolites and the associated metabolic pathways in the treated mice. The common targets of viral pneumonia and pulmonary fibrosis were acquired from the publicly available databases, and the core targets and active constituents were screened using the protein-protein interaction (PPI) network, GO and KEGG enrichment analyses, and molecular docking, and a "gene-metabolite" regulatory network was constructed. The expressions of the core targets were detected in the lung tissues of the treated mice using Western blotting. Results Fifty-three chemical constituents were identified from Arctium lappa extract. In the mouse models of pulmonary fibrosis, treatment with Arctium lappa extract significantly improved weight loss and ameliorated lung inflammation and fibrosis. The differential metabolites in the treated mice were enriched in energy metabolism pathways involving citrate cycle, pentose phosphate pathway, glycolysis, tryptophan metabolism, glutamate metabolism and glutathione metabolism, which regulated the production of energy metabolism intermediates. Twenty-three key active compounds (mostly lignans and phenolic acids) and 82 core targets were screened, which were associated with the non-canonical Smad signaling pathways (including PI3K/AKT, HIF-1, MAPK, and Foxo) that participated in the regulation of energy metabolism. Arctium lappa extract also regulated the expressions of epithelial-mesenchymal transition (EMT)‑related proteins (fibronectin, vimentim, and Snail, etc.) and inhibited MAPK signaling pathway activation. Conclusion Preliminary findings suggest that Arctium lappa treats fibrosis by regulating metabolism to inhibit EMT and involves the modulation of non-canonical Smad signaling pathways, such as MAPK providing theoretical support for its clinical application and further research in treating PPF.

Key words: Arctium lappa, post-viral pneumonia pulmonary fibrosis, network pharmacology, metabolomics, MAPK

Guoyong LI, Renling LI, Yiting LIU, Hongxia KE, Jing LI, Xinhua WANG. Therapeutic mechanism of Arctium lappa extract for post-viral pneumonia pulmonary fibrosis: a metabolomics, network pharmacology analysis and experimental verification[J]. Journal of Southern Medical University, 2025, 45(6): 1185-1199.

Figures/Tables 11

References 67

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No.	Retention time (min)	m/z			Adduct ion	Molecular formula	Compound	Fragment ion
No.	Retention time (min)	Theoretical	Measured	Mass error/ppm	Adduct ion	Molecular formula	Compound	Fragment ion
1	1.574	138.055	138.0554	3.39	[M+H]⁺	C₃H₇NO₂	Aminobenzoic acid ^[15]	120.0449, 94.0657
2	2.228	268.104	268.1044	1.69	[M+H]⁺	C₁₀H₁₃N₅O₄	Adenosine ^[16]	136.0608, 119.0346
3	2.494	132.1019	132.1023	2.93	[M+H]⁺	C₆H₁₃NO₂	L-Leucine ^[17]	86.0972, 69.0705
4	2.686	132.1019	132.1023	3.08	[M+H]⁺	C₆H₁₃NO₂	L-isoleucine ^[17]	86.0965, 77.0386
5	2.854	331.0671	331.0753	3.02	[M-H]^-	C₁₃H₁₆O₁₀	1-O-Galloyl-glucose ^[18]	168.0063, 125.0242
6	3.237	345.0827	345.091	3.01	[M-H]^-	C₁₄H₁₈O₁₀	Methyl-6-O-galloyl-β-D-glucopyranoside ^[19]	137.0249, 93.0347
7	3.802	166.0863	166.0868	3.14	[M+H]⁺	C₉H₁₁NO₂	L-Phenylalanine ^[20]	120.0811, 103.0544
8	3.919	447.1508	447.1504	2.34	[M-H]^-	C₁₉H₂₈O₁₂	4-hydroxy-3-methoxybenzylalcohol-4-O-β-D-xylopyranosyl-(1→6)-O-β-D-glucopyranoside ^[21]	293.0881, 149.0456
9	4.634	353.0878	353.0888	2.53	[M-H]^-	C₁₆H₁₈O₉	5-caffeoylquinic acid^[22]	135.0453, 191.0562
10	4.932	113.0597	113.0602	4.42	[M+H]⁺	C₆H₈O₂	Parasorbic acid^[23]	95.0496, 59.0501
11	5.199	205.0972	205.0974	1.36	[M+H]⁺	C₁₁H₁₂N₂O₂	Tryptophan^[20]	146.0610, 118.0657
12	6.431	353.0878	353.0891	3.32	[M-H]^-	C₁₆H₁₈O₉	Chlorogenic acid(NBZ1)^[22]	191.0569, 127.0402
13	6.681	353.0878	353.0890	3.24	[M-H]^-	C₁₆H₁₈O₉	Cryptochlorgenic acid^[22]	191.0571, 85.0301
14	7.163	537.1978	537.1987	0.88	[M-H]^-	C₂₆H₃₄O₁₂	Terebinthacoside III(NBZ2)^[24]	327.1251, 312.0999
15	7.496	179.035	179.0357	4.02	[M-H]^-	C₉H₈O₄	Caffeic acid(NBZ3)^[25]	179.0351, 149.0611
16	7.579	161.0244	161.0246	4.97	[M-H]^-	C₉H₆O₃	Hydroxycoumarin(NBZ4)^[26]	177.0562, 133.0660
17	8.061	337.0929	337.0941	3.26	[M-H]^-	C₁₆H₁₈O₈	3-p-Coumaroylquinic acid ^[27]	191.0568, 93.0346
18	8.311	539.2134	539.2079	1.22	[M-H]^-	C₂₆H₃₆O₁₂	(7R,8R)-7,9,9'-trihydroxy-3,3'-dimethoxy-8-O-4'-neolignan-9'-O-β-D-glucopyranoside(NBZ5)^[21]	377.1613, 195.0668
19	9.409	681.24	681.2461	-1.69	[M-H]^-	C₃₂H₄₂O₁₆	Pinoresinol-4'4'-O-β-D-glucoside^[21]	519.1874, 357.1347
20	10.326	417.1755	417.1865	1.41	[M+H]⁺	C₁₉H₂₈O₁₀	2-phenethyl-β-primeveroside^[21]	307.1153, 159.0806
21	10.907	567.2083	567.2092	0.85	[M+HCOO]^-	C₂₆H₃₄O₁₁	Lariciresinol-4-β-D-glucopyranoside^[28]	151.0394, 314.1152
22	11.672	535.1821	535.1780	1	[M-H]^-	C₂₆H₃₂O₁₂	4,4',9'-trihydroxy-3,3'-dimethoxy-7',9-epoxylignan-7-oxo-4-O-β-D-glucopyranoside^[21]	387.1451, 151.0403
23	12.105	515.1195	515.1207	1.98	[M-H]^-	C₂₅H₂₄O₁₂	3,5-dicaffeoylquinic acid(NBZ6)^[29]	353.0881, 191.0562
24	12.347	749.2815	749.2821	0.79	[M-H]^-	C₄₀H₄₆O₁₄	Lappaol H(NBZ7)^[38]	665.2400, 559.1979
25	12.720	549.1978	549.1972	-1.55	[M-H]^-	C₂₇H₃₄O₁₂	2-hydroxyarctiin ^[31]	387.1456, 151.0406
26	13.363	519.1872	519.1868	-0.97	[M-H]^-	C₂₆H₃₂O₁₁	Pinoresinol‑O‑β‑D‑glucopyranoside(NBZ8)^[32]	357.1348, 221.0817
27	14.334	515.1195	515.1205	1.7	[M-H]^-	C₂₅H₂₄O₁₂	4,5-dicaffeoylquinic acid(NBZ9)^[29]	353.0898, 173.0463
28	14.983	553.2079	553.2087	1.06	[M-H]^-	C₃₀H₃₄O₁₀	Lappaol C(NBZ10)^[33]	505.1874, 411.1458
29	15.166	553.2079	553.2087	0.91	[M-H]^-	C₃₀H₃₄O₁₀	Isolappaol C(NBZ11)^[34]	505.1874, 411.1458
30	15.796	552.2439	552.2442	2.1	[M+NH₄]⁺	C₂₇H₃₄O₁₁	Arctiin(NBZ12)^[35]	373.1655, 355.1547
31	17.079	553.2068	553.2077	1.63	[M+H]⁺	C₃₀H₃₂O₁₀	Arctignan C^[36]	505.1858, 353.0384
32	17.379	551.1923	551.1930	1	[M-H]^-	C₃₀H₃₂O₁₀	Arctignan B^[37]	521.1819, 397.1295
33	17.761	731.2709	731.2712	0.32	[M-H]^-	C₄₀H₄₄O₁₃	Arctignan D(NBZ13)^[38]	665.2398, 559.1972
34	17.911	553.2079	553.2087	1.26	[M-H]^-	C₃₀H₃₄O₁₀	Arctignan A(NBZ14)^[37]	505.1875, 357.1352
35	17.928	357.1344	357.1354	2.62	[M-H]^-	C₂₀H₂₂O₆	Matairesinol(NBZ15)^[34]	342.1115, 221.0825
36	17.994	567.2236	567.2244	0.98	[M-H]^-	C₃₁H₃₆O₁₀	Lappaol D(NBZ16)^[39]	415.1764, 519.2021
37	18.028	731.2709	731.2713	0.48	[M-H]^-	C₄₀H₄₄O₁₃	Arctignan E(NBZ17)^[38]	559.1964, 665.2379
38	20.307	559.1939	559.1949	1.94	[M+Na]⁺	C₃₀H₃₂O₉	Lappaol A(NBZ18)^[30]	460.0799, 137.0597
39	20.581	537.2119	537.2075	1.7	[M+H]⁺	C₃₀H₃₂O₉	Isolappaol A(NBZ19^[34])	371.1481 137.0599
40	21.039	373.1646	373.1644	0.96	[M+H]⁺	C₂₁H₂₄O₆	Arctigenin(NBZ20)^[40]	305.1175 137.0602
41	22.137	713.2604	713.2609	0.5	[M-H]^-	C₄₀H₄₂O₁₂	Lappaol F(NBZ21)^[33]	665.2381, 653.2382
42	22.753	549.213	549.2134	0.5	[M-H]^-	C₃₁H₃₄O₉	Lappaol B(NBZ22)^[33]	531.2001, 519.2020
43	34.532	313.2463	313.2392	2.81	[M+CH3COO]^-	C₁₆H₃₀O₂	Hexadecenoic acid ^[41]	183.1394, 129.0921
44	35.281	315.2541	315.2548	2.42	[M+HCOO]^-	C₁₇H₃₄O₂	Daturic acid ^[42]	112.9852, 127.1120
45	35.647	295.2279	295.2278	0.52	[M-H]^-	C₁₈H₃₂O₃	9(10)-EpOME^[43]	277.2171, 195.1386
46	36.795	297.2435	297.2444	2.47	[M-H]^-	C₁₈H₃₄O₃	9-hydroxy-12-octadecenoic acid ^[44]	279.2327, 127.1117
47	37.66	277.2173	277.2181	2.69	[M-H]^-	C₁₈H₃₀O₂	Linolenic acid ^[20]	233.2273, 177.1655
48	37.712	353.2662	353.2673	2.68	[M+Na]⁺	C₁₉H₃₈O₄	Monopalmitin ^[45]	209.1541, 91.0553
49	38.093	279.233	279.2338	3.05	[M-H]^-	C₁₈H₃₂O₂	Linoleicacid(NBZ23)^[20]	261.2213, 167.0723
50	38.675	255.233	255.2337	2.41	[M-H]^-	C₁₆H₃₂O₂	Methyl pentadecanoate ^[46]	209.0621, 108.0204
51	39.041	281.2486	281.2492	1.98	[M-H]^-	C₁₈H₃₄O₂	Oleic acid ^[20]	127.0748, 263.2380
52	39.061	376.3421	376.3403	2.12	[M+NH4]⁺	C₂₁H₄₂O₄	2-monostearin ^[40]	260.2363, 95.0850
53	40.055	283.2643	283.2645	0.67	[M-H]^-	C₁₈H₃₆O₂	Octadecanoic acid ^[20]	96.0190, 161.0050

No.	Retention time (min)	m/z			Adduct ion	Molecular formula	Compound	Fragment ion
No.	Retention time (min)	Theoretical	Measured	Mass error/ppm	Adduct ion	Molecular formula	Compound	Fragment ion
1	1.574	138.055	138.0554	3.39	[M+H]⁺	C₃H₇NO₂	Aminobenzoic acid ^[15]	120.0449, 94.0657
2	2.228	268.104	268.1044	1.69	[M+H]⁺	C₁₀H₁₃N₅O₄	Adenosine ^[16]	136.0608, 119.0346
3	2.494	132.1019	132.1023	2.93	[M+H]⁺	C₆H₁₃NO₂	L-Leucine ^[17]	86.0972, 69.0705
4	2.686	132.1019	132.1023	3.08	[M+H]⁺	C₆H₁₃NO₂	L-isoleucine ^[17]	86.0965, 77.0386
5	2.854	331.0671	331.0753	3.02	[M-H]^-	C₁₃H₁₆O₁₀	1-O-Galloyl-glucose ^[18]	168.0063, 125.0242
6	3.237	345.0827	345.091	3.01	[M-H]^-	C₁₄H₁₈O₁₀	Methyl-6-O-galloyl-β-D-glucopyranoside ^[19]	137.0249, 93.0347
7	3.802	166.0863	166.0868	3.14	[M+H]⁺	C₉H₁₁NO₂	L-Phenylalanine ^[20]	120.0811, 103.0544
8	3.919	447.1508	447.1504	2.34	[M-H]^-	C₁₉H₂₈O₁₂	4-hydroxy-3-methoxybenzylalcohol-4-O-β-D-xylopyranosyl-(1→6)-O-β-D-glucopyranoside ^[21]	293.0881, 149.0456
9	4.634	353.0878	353.0888	2.53	[M-H]^-	C₁₆H₁₈O₉	5-caffeoylquinic acid^[22]	135.0453, 191.0562
10	4.932	113.0597	113.0602	4.42	[M+H]⁺	C₆H₈O₂	Parasorbic acid^[23]	95.0496, 59.0501
11	5.199	205.0972	205.0974	1.36	[M+H]⁺	C₁₁H₁₂N₂O₂	Tryptophan^[20]	146.0610, 118.0657
12	6.431	353.0878	353.0891	3.32	[M-H]^-	C₁₆H₁₈O₉	Chlorogenic acid(NBZ1)^[22]	191.0569, 127.0402
13	6.681	353.0878	353.0890	3.24	[M-H]^-	C₁₆H₁₈O₉	Cryptochlorgenic acid^[22]	191.0571, 85.0301
14	7.163	537.1978	537.1987	0.88	[M-H]^-	C₂₆H₃₄O₁₂	Terebinthacoside III(NBZ2)^[24]	327.1251, 312.0999
15	7.496	179.035	179.0357	4.02	[M-H]^-	C₉H₈O₄	Caffeic acid(NBZ3)^[25]	179.0351, 149.0611
16	7.579	161.0244	161.0246	4.97	[M-H]^-	C₉H₆O₃	Hydroxycoumarin(NBZ4)^[26]	177.0562, 133.0660
17	8.061	337.0929	337.0941	3.26	[M-H]^-	C₁₆H₁₈O₈	3-p-Coumaroylquinic acid ^[27]	191.0568, 93.0346
18	8.311	539.2134	539.2079	1.22	[M-H]^-	C₂₆H₃₆O₁₂	(7R,8R)-7,9,9'-trihydroxy-3,3'-dimethoxy-8-O-4'-neolignan-9'-O-β-D-glucopyranoside(NBZ5)^[21]	377.1613, 195.0668
19	9.409	681.24	681.2461	-1.69	[M-H]^-	C₃₂H₄₂O₁₆	Pinoresinol-4'4'-O-β-D-glucoside^[21]	519.1874, 357.1347
20	10.326	417.1755	417.1865	1.41	[M+H]⁺	C₁₉H₂₈O₁₀	2-phenethyl-β-primeveroside^[21]	307.1153, 159.0806
21	10.907	567.2083	567.2092	0.85	[M+HCOO]^-	C₂₆H₃₄O₁₁	Lariciresinol-4-β-D-glucopyranoside^[28]	151.0394, 314.1152
22	11.672	535.1821	535.1780	1	[M-H]^-	C₂₆H₃₂O₁₂	4,4',9'-trihydroxy-3,3'-dimethoxy-7',9-epoxylignan-7-oxo-4-O-β-D-glucopyranoside^[21]	387.1451, 151.0403
23	12.105	515.1195	515.1207	1.98	[M-H]^-	C₂₅H₂₄O₁₂	3,5-dicaffeoylquinic acid(NBZ6)^[29]	353.0881, 191.0562
24	12.347	749.2815	749.2821	0.79	[M-H]^-	C₄₀H₄₆O₁₄	Lappaol H(NBZ7)^[38]	665.2400, 559.1979
25	12.720	549.1978	549.1972	-1.55	[M-H]^-	C₂₇H₃₄O₁₂	2-hydroxyarctiin ^[31]	387.1456, 151.0406
26	13.363	519.1872	519.1868	-0.97	[M-H]^-	C₂₆H₃₂O₁₁	Pinoresinol‑O‑β‑D‑glucopyranoside(NBZ8)^[32]	357.1348, 221.0817
27	14.334	515.1195	515.1205	1.7	[M-H]^-	C₂₅H₂₄O₁₂	4,5-dicaffeoylquinic acid(NBZ9)^[29]	353.0898, 173.0463
28	14.983	553.2079	553.2087	1.06	[M-H]^-	C₃₀H₃₄O₁₀	Lappaol C(NBZ10)^[33]	505.1874, 411.1458
29	15.166	553.2079	553.2087	0.91	[M-H]^-	C₃₀H₃₄O₁₀	Isolappaol C(NBZ11)^[34]	505.1874, 411.1458
30	15.796	552.2439	552.2442	2.1	[M+NH₄]⁺	C₂₇H₃₄O₁₁	Arctiin(NBZ12)^[35]	373.1655, 355.1547
31	17.079	553.2068	553.2077	1.63	[M+H]⁺	C₃₀H₃₂O₁₀	Arctignan C^[36]	505.1858, 353.0384
32	17.379	551.1923	551.1930	1	[M-H]^-	C₃₀H₃₂O₁₀	Arctignan B^[37]	521.1819, 397.1295
33	17.761	731.2709	731.2712	0.32	[M-H]^-	C₄₀H₄₄O₁₃	Arctignan D(NBZ13)^[38]	665.2398, 559.1972
34	17.911	553.2079	553.2087	1.26	[M-H]^-	C₃₀H₃₄O₁₀	Arctignan A(NBZ14)^[37]	505.1875, 357.1352
35	17.928	357.1344	357.1354	2.62	[M-H]^-	C₂₀H₂₂O₆	Matairesinol(NBZ15)^[34]	342.1115, 221.0825
36	17.994	567.2236	567.2244	0.98	[M-H]^-	C₃₁H₃₆O₁₀	Lappaol D(NBZ16)^[39]	415.1764, 519.2021
37	18.028	731.2709	731.2713	0.48	[M-H]^-	C₄₀H₄₄O₁₃	Arctignan E(NBZ17)^[38]	559.1964, 665.2379
38	20.307	559.1939	559.1949	1.94	[M+Na]⁺	C₃₀H₃₂O₉	Lappaol A(NBZ18)^[30]	460.0799, 137.0597
39	20.581	537.2119	537.2075	1.7	[M+H]⁺	C₃₀H₃₂O₉	Isolappaol A(NBZ19^[34])	371.1481 137.0599
40	21.039	373.1646	373.1644	0.96	[M+H]⁺	C₂₁H₂₄O₆	Arctigenin(NBZ20)^[40]	305.1175 137.0602
41	22.137	713.2604	713.2609	0.5	[M-H]^-	C₄₀H₄₂O₁₂	Lappaol F(NBZ21)^[33]	665.2381, 653.2382
42	22.753	549.213	549.2134	0.5	[M-H]^-	C₃₁H₃₄O₉	Lappaol B(NBZ22)^[33]	531.2001, 519.2020
43	34.532	313.2463	313.2392	2.81	[M+CH3COO]^-	C₁₆H₃₀O₂	Hexadecenoic acid ^[41]	183.1394, 129.0921
44	35.281	315.2541	315.2548	2.42	[M+HCOO]^-	C₁₇H₃₄O₂	Daturic acid ^[42]	112.9852, 127.1120
45	35.647	295.2279	295.2278	0.52	[M-H]^-	C₁₈H₃₂O₃	9(10)-EpOME^[43]	277.2171, 195.1386
46	36.795	297.2435	297.2444	2.47	[M-H]^-	C₁₈H₃₄O₃	9-hydroxy-12-octadecenoic acid ^[44]	279.2327, 127.1117
47	37.66	277.2173	277.2181	2.69	[M-H]^-	C₁₈H₃₀O₂	Linolenic acid ^[20]	233.2273, 177.1655
48	37.712	353.2662	353.2673	2.68	[M+Na]⁺	C₁₉H₃₈O₄	Monopalmitin ^[45]	209.1541, 91.0553
49	38.093	279.233	279.2338	3.05	[M-H]^-	C₁₈H₃₂O₂	Linoleicacid(NBZ23)^[20]	261.2213, 167.0723
50	38.675	255.233	255.2337	2.41	[M-H]^-	C₁₆H₃₂O₂	Methyl pentadecanoate ^[46]	209.0621, 108.0204
51	39.041	281.2486	281.2492	1.98	[M-H]^-	C₁₈H₃₄O₂	Oleic acid ^[20]	127.0748, 263.2380
52	39.061	376.3421	376.3403	2.12	[M+NH4]⁺	C₂₁H₄₂O₄	2-monostearin ^[40]	260.2363, 95.0850
53	40.055	283.2643	283.2645	0.67	[M-H]^-	C₁₈H₃₆O₂	Octadecanoic acid ^[20]	96.0190, 161.0050

PDB ID	4GV1 6NJS 1US0 6GR5 3HHM
PDB ID	AKT1	STAT3	SRC	HSP90AA1	PIK3CA
Arctigenin	-6.507	-6.402	-5.763	-6.547	-7.426
Isolappaol A	-8.067	-8.169	-7.637	-6.55	-9.424
Lappaol A	-6.757	-8.544	-7.251	-6.45	-9.126
Lappaol D	-7.564	-5.689	-7.099	-5.803	-7.943
Matairesinol	-6.829	-5.633	-6.700	-6.764	-6.998

PDB ID	4GV1 6NJS 1US0 6GR5 3HHM
PDB ID	AKT1	STAT3	SRC	HSP90AA1	PIK3CA
Arctigenin	-6.507	-6.402	-5.763	-6.547	-7.426
Isolappaol A	-8.067	-8.169	-7.637	-6.55	-9.424
Lappaol A	-6.757	-8.544	-7.251	-6.45	-9.126
Lappaol D	-7.564	-5.689	-7.099	-5.803	-7.943
Matairesinol	-6.829	-5.633	-6.700	-6.764	-6.998

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