2.1 Results of active ingredients of Panax notoginseng, Salvia miltiorrhiza Bge., and Borneol
According to the qualification conditions, 6 active ingredients of Panax notoginseng, 42 active ingredients of Salvia miltiorrhiza and 3 active ingredients of Borneol were collected from TCMSP database. (Table 1)
Table 1 Active ingredients of CDDP
Chinese medicine
|
Compounds
|
OB (%)
|
DL
|
PubChem ID
|
Panax notoginseng
|
quercetin
|
46.43334812
|
0.27525
|
5280343
|
|
beta-sitosterol
|
36.91390583
|
0.75123
|
222284
|
|
Stigmasterol
|
43.82985158
|
0.75665
|
5280794
|
|
Mandenol
|
41.99620045
|
0.19321
|
5282184
|
|
ginsenoside rh2
|
36.31951
|
0.55868
|
119307
|
|
ginsenoside f2
|
36.43175
|
0.25282
|
9918692
|
Salvia miltiorrhiza Bge.
|
tanshinone iia
|
49.88730004
|
0.39781
|
164676
|
|
Tanshindiol B
|
42.66581049
|
0.45303
|
5321620
|
|
tanshinaldehyde
|
52.4747043
|
0.45196
|
124268
|
|
sugiol
|
36.11353486
|
0.27648
|
94162
|
|
sclareol
|
43.67068458
|
0.2058
|
163263
|
|
salviolone
|
31.72415039
|
0.23568
|
10355691
|
|
Salvilenone
|
30.38365387
|
0.37639
|
389885
|
|
salvianolic acid j
|
43.37604991
|
0.72497
|
24177556
|
|
salvianolic acid g
|
45.56485578
|
0.60602
|
11530200
|
|
przewaquinone f
|
40.30788399
|
0.45925
|
126073
|
|
Przewaquinone E
|
42.85485204
|
0.45301
|
126072
|
|
przewaquinone c
|
55.7416731
|
0.40408
|
126071
|
|
Przewaquinone B
|
62.24005962
|
0.41374
|
622085
|
|
przewalskin b
|
110.3240001
|
0.43809
|
16102114
|
|
przewalskin a
|
37.10650066
|
0.64901
|
16090911
|
|
Poriferasterol
|
43.82985158
|
0.75596
|
5281330
|
|
poriferast-5-en-3beta-ol
|
36.91390583
|
0.75034
|
457801
|
|
neocryptotanshinone ii
|
39.46299114
|
0.23157
|
15690458
|
|
neocryptotanshinone
|
52.48799701
|
0.32306
|
389888
|
|
Miltirone
|
38.75698635
|
0.25418
|
160142
|
|
miltipolone
|
36.55611206
|
0.36803
|
10086184
|
|
miltionone Ⅱ
|
71.02970321
|
0.43711
|
51531511
|
|
miltionone Ⅰ
|
49.68439433
|
0.32125
|
53474332
|
|
microstegiol
|
39.61229457
|
0.27734
|
403772
|
|
Methylenetanshinquinone
|
37.07319368
|
0.36017
|
105118
|
|
manool
|
45.04431636
|
0.20208
|
3034394
|
|
luteolin
|
36.16262934
|
0.24552
|
5280445
|
|
Isotanshinone II
|
49.91602574
|
0.39674
|
44425166
|
|
isoimperatorin
|
45.46424674
|
0.22524
|
68081
|
|
formyltanshinone
|
73.444622
|
0.41736
|
348675638
|
|
epidanshenspiroketallactone
|
68.27315929
|
0.30549
|
102004791
|
|
dihydrotanshinoneⅠ
|
45.04327919
|
0.36015
|
11425923
|
|
digallate
|
61.84861803
|
0.25635
|
54711004
|
|
Dehydrotanshinone II A
|
43.76228599
|
0.40019
|
128994
|
|
Danshenol B
|
57.9508753
|
0.55764
|
3083515
|
|
Danshenol A
|
56.96524899
|
0.52172
|
3083514
|
|
cryptotanshinone
|
52.34196226
|
0.39555
|
160254
|
|
C09092
|
36.06948986
|
0.2474
|
442027
|
|
Baicalin
|
40.12360996
|
0.75264
|
64982
|
|
4-methylenemiltirone
|
34.34867589
|
0.22726
|
14609851
|
|
2-isopropyl-8-methylphenanthrene-3,4-dione
|
40.86015408
|
0.22897
|
135872
|
|
1,2,5,6-tetrahydrotanshinone
|
38.74538672
|
0.35791
|
124416
|
Borneol
|
Asiatic acid
|
41.38281219
|
0.71097
|
119034
|
|
Bronyl acetate
|
59.29526304
|
0.51159
|
93009
|
|
Dipterocarpol
|
41.708061
|
0.76437
|
441676
|
2.2 CDDP-Compounds-Targets-DR Network
In TCMSP, there are 186 targets of Panax notoginseng, 215 targets of Salvia miltiorrhiza Bge., and without validated targets for Borneol. There are 433 predict targets for Panax notoginseng, 2850 predict targets for Salvia miltiorrhiza Bge., and 246 predict targets for Borneol. The ingredients of CDDP and their corresponding targets were introduced into the Cytoscape software, and CDDP-Compunds-Targets-DR Network was drawn. (Figure 2)
2.3 Disease and drug target intersection results
Using MCODE cluster analysis, the results showed that there were two clusters A and B, a total of 64 targets, including ABCB1, ACE, AGTR1. Cluster A: score 26.486, nodes 38, edges 490. Cluster B: score 9.6, nodes 26, edges 490. (Figure 3)
The String database was used to display the interaction between proteins, in which the red line represents fusion evidence, the green line represents adjacent evidence, the blue line represents coexistence evidence, the purple line represents experimental evidence, the yellow line represents text mining evidence, the light blue line represents database evidence, and the black line represents Co expression evidence. The results show that, number of nodes: 64. number of edges: 1100. average node degree: 34.4. avg. local clustering coefficient: 0.769. expected number of edges: 276. PPI enrichment p-value: < 1.0e-16. (Figure 4)
2.4 Signal pathway enrichment results
According to the results obtained from DAVID database, the top 20 of KEGG pathway mainly include TNF signaling pathway and HIF-1 signaling pathway. In GO analysis, BP mainly includes positive regulation of smooth muscle cell proliferation and response to hypoxia, CC mainly includes extracellular space and extracellular domain, MF mainly includes protein binding and protein binding recognition. (Figure 5)
In KEGG database, we searched the TNF signaling pathway, HIF-1 signaling pathway and the key genes in the pathway, and preliminarily determined that TNF, NFkB and VEGF were the key genes in TNF signaling pathway, while IL-6, STAT3, HIF1A and VEGF were the key genes in HIF-1 signaling pathway. (Figure 6, 7)
2.5 Molecular docking results
In KEGG database, we preliminarily determined that TNF, NFkB and VEGF are the key genes in TNF signaling pathway, while IL-6, STAT3, HIF1A and VEGF are the key genes in HIF-1 signaling pathway. In this regard, we used 51 compounds from CDDP to obtain the 2D structures of 46 compounds in PubChem database, and docking with TNF, NFkB, VEGF, IL-6, STAT3, HIF1A molecules respectively.
Molecular docking results showed that all components of CDDP had a certain docking ability with TNF, NFkB, VEGF, IL-6, STAT3 and HIF1A, among which Asiatic acid (PubChem ID 119034) and Salvianolic acid j (PubChem ID 24177556) had the strongest docking ability (Figure 8). Among them, the first three substances docking with TNF were Asiatic acid, Ginsenoside f2 and Danshenol B. The first three substances docking with NFkB were Asiatic acid, Przewalskin b and Salvianolic acid j. The first three substances docking with VEGF were Asiatic acid, Ginsenoside f2, Salvianolic acid j. The first three substances docking with IL-6 were Asiatic acid, Danshenol B and Przewalskin b. The first three substances docking with STAT3 were Asiatic acid, Ginsenoside f2 and Salvianolic acid j. The first three substances docking with HIF1A were Asiatic acid, Ginsenoside f2 and Salvianolic acid j. (Figure 9)