Active ingredients of Xiaochaihu Decoction
There were 196 active ingredients of 7 herbs containing in Xiaochaihu Decoction which were collected from TCMSP Database with the limited lists “OB ≥ 30%, OL ≥ 0.18”. As is shown in Table.2, the active composition included 17 in Chaihu, 13 in Banxia, 36 in Huangqin, 22 in Renshen, 29 in Daozao, 5 in shenjiang, 94 in Gancao.
Table.2
Active ingredients information of Xiaochaihu Decoction
Mol ID
|
Molecule Name
|
OB (%)
|
DL
|
Herb
|
MOL000073
|
ent-Epicatechin
|
48.96
|
0.24
|
Scutellariae Radix
|
MOL000096
|
(-)-catechin
|
49.68
|
0.24
|
Jujubae Fructus
|
MOL000098
|
quercetin
|
46.43
|
0.28
|
Radix Bupleuri、licorice、Jujubae Fructus
|
MOL000173
|
wogonin
|
30.68
|
0.23
|
Scutellariae Radix
|
MOL000211
|
Mairin
|
55.38
|
0.78
|
licorice、Jujubae Fructus
|
MOL000228
|
(2R)-7-hydroxy-5-methoxy-2-phenylchroman-4-one
|
55.23
|
0.2
|
Scutellariae Radix
|
MOL000239
|
Jaranol
|
50.83
|
0.29
|
licorice
|
MOL000354
|
isorhamnetin
|
49.6
|
0.31
|
Radix Bupleuri、licorice
|
MOL000358
|
beta-sitosterol
|
36.91
|
0.75
|
Arum Ternatum Thunb、Panax ginseng C. A. Mey.、Scutellariae Radix、Zingiber officinale Roscoe、Jujubae Fructus
|
MOL000359
|
sitosterol
|
36.91
|
0.75
|
Scutellariae Radix、licorice
|
MOL000392
|
formononetin
|
69.67
|
0.21
|
licorice
|
MOL000417
|
Calycosin
|
47.75
|
0.24
|
licorice
|
MOL000422
|
kaempferol
|
41.88
|
0.24
|
Radix Bupleuri、Panax ginseng C. A. Mey.、licorice
|
MOL000449
|
Stigmasterol
|
43.83
|
0.76
|
Arum Ternatum Thunb、Panax ginseng C. A. Mey.、Scutellariae Radix、Zingiber officinale Roscoe、Jujubae Fructus、Radix Bupleuri
|
MOL000490
|
petunidin
|
30.05
|
0.31
|
Radix Bupleuri
|
MOL000492
|
(+)-catechin
|
54.83
|
0.24
|
Jujubae Fructus
|
MOL000497
|
licochalcone a
|
40.79
|
0.29
|
licorice
|
MOL000500
|
licochalcone a
|
40.79
|
0.29
|
licorice
|
MOL000500
|
Vestitol
|
74.66
|
0.21
|
licorice
|
MOL000519
|
coniferin
|
31.11
|
0.32
|
Arum Ternatum Thunb
|
MOL000525
|
Norwogonin
|
39.4
|
0.21
|
Scutellariae Radix
|
MOL000552
|
5,2'-Dihydroxy-6,7,8-trimethoxyflavone
|
31.71
|
0.35
|
Scutellariae Radix
|
MOL000627
|
Stepholidine
|
33.11
|
0.54
|
Jujubae Fructus
|
MOL000783
|
Protoporphyrin
|
30.86
|
0.56
|
Jujubae Fructus
|
MOL000787
|
Fumarine
|
59.26
|
0.83
|
Panax ginseng C. A. Mey.、Jujubae Fructus
|
MOL001454
|
berberine
|
36.86
|
0.78
|
Jujubae Fructus
|
MOL001458
|
coptisine
|
30.67
|
0.86
|
Scutellariae Radix
|
MOL001484
|
Inermine
|
75.18
|
0.54
|
licorice
|
MOL001490
|
bis[(2S)-2-ethylhexyl] benzene-1,2-dicarboxylate
|
43.59
|
0.35
|
Scutellariae Radix
|
MOL001506
|
Supraene
|
33.55
|
0.42
|
Scutellariae Radix
|
MOL001522
|
(S)-Coclaurine
|
42.35
|
0.24
|
Jujubae Fructus
|
MOL001645
|
Linoleyl acetate
|
42.1
|
0.2
|
Radix Bupleuri
|
MOL001689
|
acacetin
|
34.97
|
0.24
|
Scutellariae Radix
|
MOL001755
|
24-Ethylcholest-4-en-3-one
|
36.08
|
0.76
|
Arum Ternatum Thunb
|
MOL001771
|
poriferast-5-en-3beta-ol
|
36.91
|
0.75
|
Zingiber officinale Roscoe
|
MOL001792
|
DFV
|
32.76
|
0.18
|
licorice
|
MOL002311
|
Glycyrol
|
90.78
|
0.67
|
licorice
|
MOL002565
|
Medicarpin
|
49.22
|
0.34
|
licorice
|
MOL002670
|
Cavidine
|
35.64
|
0.81
|
Arum Ternatum Thunb
|
MOL002714
|
baicalein
|
33.52
|
0.21
|
Arum Ternatum Thunb、Scutellariae Radix
|
MOL002773
|
beta-carotene
|
37.18
|
0.58
|
Jujubae Fructus
|
MOL002776
|
Baicalin
|
40.12
|
0.75
|
Arum Ternatum Thunb、Radix Bupleuri
|
MOL002844
|
Pinocembrin
|
64.72
|
0.18
|
licorice
|
MOL002879
|
Diop
|
43.59
|
0.39
|
Panax ginseng C. A. Mey.、Scutellariae Radix
|
MOL002897
|
epiberberine
|
43.09
|
0.78
|
Scutellariae Radix
|
MOL002908
|
5,8,2'-Trihydroxy-7-methoxyflavone
|
37.01
|
0.27
|
Scutellariae Radix
|
MOL002909
|
5,7,2,5-tetrahydroxy-8,6-dimethoxyflavone
|
33.82
|
0.45
|
Scutellariae Radix
|
MOL002910
|
Carthamidin
|
41.15
|
0.24
|
Scutellariae Radix
|
MOL002911
|
2,6,2',4'-tetrahydroxy-6'-methoxychaleone
|
69.04
|
0.22
|
Scutellariae Radix
|
MOL002913
|
Dihydrobaicalin_qt
|
40.04
|
0.21
|
Scutellariae Radix
|
MOL002914
|
Eriodyctiol (flavanone)
|
41.35
|
0.24
|
Scutellariae Radix
|
MOL002915
|
Salvigenin
|
49.07
|
0.33
|
Scutellariae Radix
|
MOL002917
|
5,2',6'-Trihydroxy-7,8-dimethoxyflavone
|
45.05
|
0.33
|
Scutellariae Radix
|
MOL002925
|
5,7,2',6'-Tetrahydroxyflavone
|
37.01
|
0.24
|
Scutellariae Radix
|
MOL002926
|
dihydrooroxylin A
|
38.72
|
0.23
|
Scutellariae Radix
|
MOL002927
|
Skullcapflavone II
|
69.51
|
0.44
|
Scutellariae Radix
|
MOL002928
|
oroxylin a
|
41.37
|
0.23
|
Scutellariae Radix
|
MOL002932
|
Panicolin
|
76.26
|
0.29
|
Scutellariae Radix
|
MOL002933
|
5,7,4'-Trihydroxy-8-methoxyflavone
|
36.56
|
0.27
|
Scutellariae Radix
|
MOL002934
|
NEOBAICALEIN
|
104.34
|
0.44
|
Scutellariae Radix
|
MOL002937
|
DIHYDROOROXYLIN
|
66.06
|
0.23
|
Scutellariae Radix
|
MOL003410
|
Ziziphin_qt
|
66.95
|
0.62
|
Jujubae Fructus
|
MOL003578
|
Cycloartenol
|
38.69
|
0.78
|
Arum Ternatum Thunb
|
MOL003648
|
Inermin
|
65.83
|
0.54
|
Panax ginseng C. A. Mey.
|
MOL003656
|
Lupiwighteone
|
51.64
|
0.37
|
licorice
|
MOL003896
|
7-Methoxy-2-methyl isoflavone
|
42.56
|
0.2
|
licorice
|
MOL004328
|
naringenin
|
59.29
|
0.21
|
licorice
|
MOL004350
|
Ruvoside_qt
|
36.12
|
0.76
|
Jujubae Fructus
|
MOL004492
|
Chrysanthemaxanthin
|
38.72
|
0.58
|
Panax ginseng C. A. Mey.
|
MOL004598
|
3,5,6,7-tetramethoxy-2-(3,4,5-trimethoxyphenyl)chromone
|
31.97
|
0.59
|
Radix Bupleuri
|
MOL004609
|
Areapillin
|
48.96
|
0.41
|
Radix Bupleuri
|
MOL004624
|
Longikaurin A
|
47.72
|
0.53
|
Radix Bupleuri
|
MOL004628
|
Octalupine
|
47.82
|
0.28
|
Radix Bupleuri
|
MOL004644
|
Sainfuran
|
79.91
|
0.23
|
Radix Bupleuri
|
MOL004648
|
Troxerutin
|
31.6
|
0.28
|
Radix Bupleuri
|
MOL004653
|
(+)-Anomalin
|
46.06
|
0.66
|
Radix Bupleuri
|
MOL004702
|
saikosaponin c_qt
|
30.5
|
0.63
|
Radix Bupleuri
|
MOL004718
|
α-spinasterol
|
42.98
|
0.76
|
Radix Bupleuri
|
MOL004805
|
(2S)-2-[4-hydroxy-3-(3-methylbut-2-enyl)phenyl]-8,8-dimethyl-2,3-dihydropyrano[2,3-f]chromen-4-one
|
31.79
|
0.72
|
licorice
|
MOL004806
|
euchrenone
|
30.29
|
0.57
|
licorice
|
MOL004808
|
glyasperin B
|
65.22
|
0.44
|
licorice
|
MOL004810
|
glyasperin F
|
75.84
|
0.54
|
licorice
|
MOL004811
|
Glyasperin C
|
45.56
|
0.4
|
licorice
|
MOL004814
|
Isotrifoliol
|
31.94
|
0.42
|
licorice
|
MOL004815
|
(E)-1-(2,4-dihydroxyphenyl)-3-(2,2-dimethylchromen-6-yl)prop-2-en-1-one
|
39.62
|
0.35
|
licorice
|
MOL004820
|
kanzonols W
|
50.48
|
0.52
|
licorice
|
MOL004824
|
(2S)-6-(2,4-dihydroxyphenyl)-2-(2-hydroxypropan-2-yl)-4-methoxy-2,3-dihydrofuro[3,2-g]chromen-7-one
|
60.25
|
0.63
|
licorice
|
MOL004827
|
Semilicoisoflavone B
|
48.78
|
0.55
|
licorice
|
MOL004828
|
Glepidotin A
|
44.72
|
0.35
|
licorice
|
MOL004829
|
Glepidotin B
|
64.46
|
0.34
|
licorice
|
MOL004833
|
Phaseolinisoflavan
|
32.01
|
0.45
|
licorice
|
MOL004835
|
Glypallichalcone
|
61.6
|
0.19
|
licorice
|
MOL004838
|
8-(6-hydroxy-2-benzofuranyl)-2,2-dimethyl-5-chromenol
|
58.44
|
0.38
|
licorice
|
MOL004841
|
Licochalcone B
|
76.76
|
0.19
|
licorice
|
MOL004848
|
licochalcone G
|
49.25
|
0.32
|
licorice
|
MOL004849
|
3-(2,4-dihydroxyphenyl)-8-(1,1-dimethylprop-2-enyl)-7-hydroxy-5-methoxy-coumarin
|
59.62
|
0.43
|
licorice
|
MOL004855
|
Licoricone
|
63.58
|
0.47
|
licorice
|
MOL004856
|
Gancaonin A
|
51.08
|
0.4
|
licorice
|
MOL004857
|
Gancaonin B
|
48.79
|
0.45
|
licorice
|
MOL004860
|
licorice glycoside E
|
32.89
|
0.27
|
licorice
|
MOL004863
|
3-(3,4-dihydroxyphenyl)-5,7-dihydroxy-8-(3-methylbut-2-enyl)chromone
|
66.37
|
0.41
|
licorice
|
MOL004864
|
5,7-dihydroxy-3-(4-methoxyphenyl)-8-(3-methylbut-2-enyl)chromone
|
30.49
|
0.41
|
licorice
|
MOL004866
|
2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-6-(3-methylbut-2-enyl)chromone
|
44.15
|
0.41
|
licorice
|
MOL004879
|
Glycyrin
|
52.61
|
0.47
|
licorice
|
MOL004882
|
Licocoumarone
|
33.21
|
0.36
|
licorice
|
MOL004883
|
Licoisoflavone
|
41.61
|
0.42
|
licorice
|
MOL004884
|
Licoisoflavone B
|
38.93
|
0.55
|
licorice
|
MOL004885
|
licoisoflavanone
|
52.47
|
0.54
|
licorice
|
MOL004891
|
shinpterocarpin
|
80.3
|
0.73
|
licorice
|
MOL004898
|
(E)-3-[3,4-dihydroxy-5-(3-methylbut-2-enyl)phenyl]-1-(2,4-dihydroxyphenyl)prop-2-en-1-one
|
46.27
|
0.31
|
licorice
|
MOL004903
|
liquiritin
|
65.69
|
0.74
|
licorice
|
MOL004904
|
licopyranocoumarin
|
80.36
|
0.65
|
licorice
|
MOL004905
|
3,22-Dihydroxy-11-oxo-delta(12)-oleanene-27-alpha-methoxycarbonyl-29-oic acid
|
34.32
|
0.55
|
licorice
|
MOL004907
|
Glyzaglabrin
|
61.07
|
0.35
|
licorice
|
MOL004908
|
Glabridin
|
53.25
|
0.47
|
licorice
|
MOL004910
|
Glabranin
|
52.9
|
0.31
|
licorice
|
MOL004911
|
Glabrene
|
46.27
|
0.44
|
licorice
|
MOL004912
|
Glabrone
|
52.51
|
0.5
|
licorice
|
MOL004913
|
1,3-dihydroxy-9-methoxy-6-benzofurano[3,2-c]chromenone
|
48.14
|
0.43
|
licorice
|
MOL004914
|
1,3-dihydroxy-8,9-dimethoxy-6-benzofurano[3,2-c]chromenone
|
62.9
|
0.53
|
licorice
|
MOL004915
|
Eurycarpin A
|
43.28
|
0.37
|
licorice
|
MOL004917
|
glycyroside
|
37.25
|
0.79
|
licorice
|
MOL004924
|
(-)-Medicocarpin
|
40.99
|
0.95
|
licorice
|
MOL004935
|
Sigmoidin-B
|
34.88
|
0.41
|
licorice
|
MOL004941
|
(2R)-7-hydroxy-2-(4-hydroxyphenyl)chroman-4-one
|
71.12
|
0.18
|
licorice
|
MOL004945
|
(2S)-7-hydroxy-2-(4-hydroxyphenyl)-8-(3-methylbut-2-enyl)chroman-4-one
|
36.57
|
0.32
|
licorice
|
MOL004948
|
Isoglycyrol
|
44.7
|
0.84
|
licorice
|
MOL004949
|
Isolicoflavonol
|
45.17
|
0.42
|
licorice
|
MOL004957
|
HMO
|
38.37
|
0.21
|
licorice
|
MOL004959
|
1-Methoxyphaseollidin
|
69.98
|
0.64
|
licorice
|
MOL004961
|
Quercetin der.
|
46.45
|
0.33
|
licorice
|
MOL004966
|
3'-Hydroxy-4'-O-Methylglabridin
|
43.71
|
0.57
|
licorice
|
MOL004974
|
3'-Methoxyglabridin
|
46.16
|
0.57
|
licorice
|
MOL004978
|
2-[(3R)-8,8-dimethyl-3,4-dihydro-2H-pyrano[6,5-f]chromen-3-yl]-5-methoxyphenol
|
36.21
|
0.52
|
licorice
|
MOL004980
|
Inflacoumarin A
|
39.71
|
0.33
|
licorice
|
MOL004985
|
icos-5-enoic acid
|
30.7
|
0.2
|
licorice
|
MOL004988
|
Kanzonol F
|
32.47
|
0.89
|
licorice
|
MOL004989
|
6-prenylated eriodictyol
|
39.22
|
0.41
|
licorice
|
MOL004990
|
7,2',4'-trihydroxy-5-methoxy-3།arylcoumarin
|
83.71
|
0.27
|
licorice
|
MOL004991
|
7-Acetoxy-2-methylisoflavone
|
38.92
|
0.26
|
licorice
|
MOL004993
|
8-prenylated eriodictyol
|
53.79
|
0.4
|
licorice
|
MOL004996
|
gadelaidic acid
|
30.7
|
0.2
|
licorice
|
MOL005000
|
Gancaonin G
|
60.44
|
0.39
|
licorice
|
MOL005001
|
Gancaonin H
|
50.1
|
0.78
|
licorice
|
MOL005003
|
Licoagrocarpin
|
58.81
|
0.58
|
licorice
|
MOL005007
|
Glyasperins M
|
72.67
|
0.59
|
licorice
|
MOL005008
|
Glycyrrhiza flavonol A
|
41.28
|
0.6
|
licorice
|
MOL005012
|
Licoagroisoflavone
|
57.28
|
0.49
|
licorice
|
MOL005013
|
18α-hydroxyglycyrrhetic acid
|
41.16
|
0.71
|
licorice
|
MOL005016
|
Odoratin
|
49.95
|
0.3
|
licorice
|
MOL005017
|
Phaseol
|
78.77
|
0.58
|
licorice
|
MOL005018
|
Xambioona
|
54.85
|
0.87
|
licorice
|
MOL005020
|
dehydroglyasperins C
|
53.82
|
0.37
|
licorice
|
MOL005030
|
gondoic acid
|
30.7
|
0.2
|
Arum Ternatum Thunb
|
MOL005308
|
Aposiopolamine
|
66.65
|
0.22
|
Panax ginseng C. A. Mey.
|
MOL005314
|
Celabenzine
|
101.88
|
0.49
|
Panax ginseng C. A. Mey.
|
MOL005317
|
Deoxyharringtonine
|
39.27
|
0.81
|
Panax ginseng C. A. Mey.
|
MOL005318
|
Dianthramine
|
40.45
|
0.2
|
Panax ginseng C. A. Mey.
|
MOL005320
|
arachidonate
|
45.57
|
0.2
|
Panax ginseng C. A. Mey.
|
MOL005321
|
Frutinone A
|
65.9
|
0.34
|
Panax ginseng C. A. Mey.
|
MOL005344
|
ginsenoside rh2
|
36.32
|
0.56
|
Panax ginseng C. A. Mey.
|
MOL005348
|
Ginsenoside-Rh4_qt
|
31.11
|
0.78
|
Panax ginseng C. A. Mey.
|
MOL005356
|
Girinimbin
|
61.22
|
0.31
|
Panax ginseng C. A. Mey.
|
MOL005357
|
Gomisin B
|
31.99
|
0.83
|
Panax ginseng C. A. Mey.
|
MOL005360
|
malkangunin
|
57.71
|
0.63
|
Panax ginseng C. A. Mey.
|
MOL005360
|
malkangunin
|
57.71
|
0.63
|
Jujubae Fructus
|
MOL005376
|
Panaxadiol
|
33.09
|
0.79
|
Panax ginseng C. A. Mey.
|
MOL005384
|
suchilactone
|
57.52
|
0.56
|
Panax ginseng C. A. Mey.
|
MOL005399
|
alexandrin_qt
|
36.91
|
0.75
|
Panax ginseng C. A. Mey.
|
MOL005401
|
ginsenoside Rg5_qt
|
39.56
|
0.79
|
Panax ginseng C. A. Mey.
|
MOL006129
|
6-methylgingediacetate2
|
48.73
|
0.32
|
Zingiber officinale Roscoe
|
MOL006936
|
10,13-eicosadienoic
|
39.99
|
0.2
|
Arum Ternatum Thunb
|
MOL006937
|
12,13-epoxy-9-hydroxynonadeca-7,10-dienoic acid
|
42.15
|
0.24
|
Arum Ternatum Thunb
|
MOL006957
|
(3S,6S)-3-(benzyl)-6-(4-hydroxybenzyl)piperazine-2,5-quinone
|
46.89
|
0.27
|
Arum Ternatum Thunb
|
MOL006967
|
beta-D-Ribofuranoside, xanthine-9
|
44.72
|
0.21
|
Arum Ternatum Thunb
|
MOL007213
|
Nuciferin
|
34.43
|
0.4
|
Jujubae Fructus
|
MOL008034
|
21302-79-4
|
73.52
|
0.77
|
Jujubae Fructus
|
MOL008206
|
Moslosooflavone
|
44.09
|
0.25
|
Scutellariae Radix
|
MOL008647
|
Moupinamide
|
86.71
|
0.26
|
Jujubae Fructus
|
MOL008698
|
Dihydrocapsaicin
|
47.07
|
0.19
|
Zingiber officinale Roscoe
|
MOL010415
|
11,13-Eicosadienoic acid, methyl ester
|
39.28
|
0.23
|
Scutellariae Radix
|
MOL012245
|
5,7,4'-trihydroxy-6-methoxyflavanone
|
36.63
|
0.27
|
Scutellariae Radix
|
MOL012246
|
5,7,4'-trihydroxy-8-methoxyflavanone
|
74.24
|
0.26
|
Scutellariae Radix
|
MOL012266
|
rivularin
|
37.94
|
0.37
|
Scutellariae Radix
|
MOL012921
|
stepharine
|
31.55
|
0.33
|
Jujubae Fructus
|
MOL012940
|
Spiradine A
|
113.52
|
0.61
|
Jujubae Fructus
|
MOL012946
|
zizyphus saponin I_qt
|
32.69
|
0.62
|
Jujubae Fructus
|
MOL012961
|
jujuboside A_qt
|
36.67
|
0.62
|
Jujubae Fructus
|
MOL012976
|
coumestrol
|
32.49
|
0.34
|
Jujubae Fructus
|
MOL012980
|
Daechuine S6
|
46.48
|
0.79
|
Jujubae Fructus
|
MOL012981
|
Daechuine S7
|
44.82
|
0.83
|
Jujubae Fructus
|
MOL012986
|
Jujubasaponin V_qt
|
36.99
|
0.63
|
Jujubae Fructus
|
MOL012989
|
Jujuboside C_qt
|
40.26
|
0.62
|
Jujubae Fructus
|
MOL012992
|
Mauritine D
|
89.13
|
0.45
|
Jujubae Fructus
|
MOL013187
|
Cubebin
|
57.13
|
0.64
|
Radix Bupleuri
|
MOL013357
|
(3S,6R,8S,9S,10R,13R,14S,17R)-17-[(1R,4R)-4-ethyl-1,5-dimethylhexyl]-10,13-dimethyl-2,3,6,7,8,9,11,12,14,15,16,17-dodecahydro-1H-cyclopenta[a]phenanthrene-3,6-diol
|
34.37
|
0.78
|
Jujubae Fructus
|
Intersection targets of Xiaochaihu Decoction and Pancreatitis
Xiaochaihu Decoction had 169 targets obtained from Drugbank Database and Pancreatitis had 2344 targets collected from OMIM, GeneCards and TTD Databases. Then, Venn map showed that it had 91 intersection targets on Xiaochaihu Decoction and Pancreatitis (Figure.1).
Herb-Compound-Target-Disease Network analysis
To clarify the relationship between herb of Xiaochaihu Decoction, compound target and pancreatitis targets, the herb-compound-target-disease network was constructed by Cytoscape software. The network had 293 nodes consisted of 1 disease, 7 herbs, 194 compounds and 91 putative targets. In the terms of the relationship between compounds and targets, there were 12 compounds having targets greater than 30, and they were MOL000098 (Quercetin), MOL000422 (Kaempferol), MOL003896 (7-Methoxy-2-methyl isoflavone), MOL000787 (Fumarine), MOL000354 (Isorhamnetin), MOL000392 (formononetin), MOL002565 (Medicarpin), MOL000358 (Beta-sitosterol), MOL000449 (Stigmasterol), MOL004978(2-[(3R)-8,8-dimethyl-3,4-dihydro-2H-pyrano[6,5-f]chromen-3-yl]-5-methoxyphenol), MOL000500 (Vestitol), MOL004891 (Shinpterocarpin) acting on 75, 44, 43, 38, 34, 34, 34, 32, 31, 31, 30, 30 targets respectively. And PTGS2, HSP90A, CAMKK2, ESR1, AR, PTGS1, NOS2, NCOA2, PRSS1, F10 and SCN5A were interact with 138, 100, 97, 90, 89, 86, 83, 74, 73 and 71 compounds (Figure.2).
PPI interaction network analysis
The common targets of compounds and pancreatitis were putted into STRING database to obtain the PPI interaction network in order to provide an intuitive understand of the mechanism of Xiaochaihu Decoction acting on pancreatitis. By the analysis of STRING, it was showed that the network was composed with 91 nodes as well as 1015 edges and the average node degree was 22.3. Furthermore, network topological analysis indicated that the top 3 degrees were Mitogen-activated protein kinase 3 (MAPK3, degree=58), Interleukin - 6 (IL-6, degree=57), Tumor Protein (TP53, degree=56) which involving with the cell growth, cell apoptosis as well as inflammatory response, had the greater node degree value than other targets Figure.3.
Enrichment of functions analysis
To clarified the mechanism of Xiaochaihu Decoction on pancreatitis in detail, the Go enrichment analysis was performed by DAVID Bioinformatics Resources. Go enrichment analysis got 484 items including 363 items of Biological process (BP), 53 items of Cell Component (CC) and 68 items of Molecular Function (MF) (P<0.05) and the items which Gene count proportion greater than 10 were showed as Figure.4. Biological process was mainly related to positive regulation of transcription from RNA polymerase II promoter and Molecular Function was closely in connection with protein binding.
KEGG pathway analysis indicated that Xiaochaihu Decoction exerted its pharmacological effects on pancreatitis was closely associated with pathways in cancers (fold enrichment=5.4, P<0.001), TNF signaling way (fold enrichment=12.2, P<0.001), PI3K-Akt signaling pathway (fold enrichment=3.7, P<0.001), Influenza A (fold enrichment=7.1, P<0.001), Chagas disease (fold enrichment=11.0, P<0.001) and MAPK signaling pathway (fold enrichment=4.5, P<0.001) which were involving with MAPK3, TP53, TNF and so on (Figure.5).
Molecular docking to determine the potential targets
Three targets (IL-6, MAPK3, TP53) were selected according to the results of PPI network as the core targets of Xiaochaihu Decoction treated pancreatitis. And Herb-compound-Target-Disease network indicated Quercetin could not only interacted with 75 targets but also closely related with the three targets. Therefore, Molecular Docking technology was used to simulate the interaction between Quercetin and the three targets. The results showed that Quercetin interacted with the IL-6, MAPK3, TP53, forming Vander Waals, Carbon hydrogen bond, Pi-sulfur, Salt Bridge and so on (Figure.6).
The viability of LPS treated AR42J cells for various dose
To investigate the best concentrations of LPS to build pancreatitis model in vitro, AR42J cells viability were measured following treatment of 1, 2, 4 μg/mL LPS for 24 hours. As is shown in Figure.7 A, AR42J cells viability were achieve to nearly 60% with the treatment of LPS at the dose of 2 mg/mL for 24 hours which was for the subsequent experiment.
According to the results of Herb-Compound-Target-Disease Network analysis, Quercetin was the mainly active compound of Xiaochaihu Decoction. Quercetin (12.5, 25, 50, 100 μM) were treated with AR42J cells for 8 hours after AR42J cells were incubated with LPS (2 mg/mL) for 24 hours to detect cell viability the by MTT assay. It was showed that Quercetin at the concentrations of 25, 50 and 100 μM significantly improved the cells viability (P<0.05, 0.01), but Quercetin at dose the of 12.5 μM had less effect on it (Figure.7 B).
Quercetin inhibited the productions of IL-6, IL-1β and TNF-α
According to the PPI interaction network analysis, Xiaochaihu Decoction was closely association with inflammatory reaction. Therefore, the levels of IL-6, IL-1β and TNF-α in LPS-induced pancreatitis model in vitro were detected. LPS significantly increased the secretion of IL-1β, IL-6 and TNF-α compared with control group. At expected, Quercetin at 25, 50, 100 μM were all reduced the generations of these inflammatory factors (P<0.05, 0.01, Figure.8 A, B and C).
Quercetin downregulated the relative expression of MAPK3 and TP53
The PPI network and KEGG results all showed that Xiaochaihu Decoction functioning in the treatment of insomnia were involving with MAPK3 and TP53 which were further confirmed by Molecular docking technology. In the vitro experiment (Figure.9 A and B), MAPK3 and TP53 were both upregulated after AR42J cells treated with LPS, significantly (P<0.01). However, compared with LPS group, Quercetin obviously reduced the relative expressions MAPK3 and TP53 (P<0.05, 0.01).