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Research Article
Formula Compatibility Identification of Dachengqi
Decoction Based on the Effects of Absorbed Components in
Cerulein-Injured Pancreatic AR42J Cells
Yumei Zhang,1Lin Zhu,1,2 Jia Wang,1,3 Jianlei Zhao,1,4 Xianlin Zhao,1
Hui Guo,1Juan Li,1and Wenfu Tang1
1Sichuan Provincial Pancreatitis Center, Department of Integrative Medicine, West China Hospital, Sichuan University,
Chengdu 610041, China
2Department of Intensive Care Medicine, Sichuan Integrative Medicine Hospital, Chengdu 610041, China
3Department of General Practice, Sichuan Provincial People’s Hospital, Chengdu 610072, China
4Department of Pharmacology, School of Preclinical and Forensic Medicine, West China Medical Center, Sichuan University,
Chengdu 610041, China
Correspondence should be addressed to Wenfu Tang; hxtangwenfu@.com
Received November ; Revised January ; Accepted January
Academic Editor: Hyunsu Bae
Copyright © Yumei Zhang et al. is is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Objective. To identify the herbal formula compatibility law based on the eects of the absorbed components from DCQD on the
cerulein-injured ARJ cells. Methods. ARJ cells were pretreated for min with or without the dierent concentrations of the
absorbed components from DCQD individually or in combination or DCQD and coincubated with cerulein ( nM) for a further
h. Cell viability, lactate dehydrogenase (LDH) release,andthelevelsofapoptosisandnecrosisweremeasured.Results.Compared
to DCQD, the individual or combination components partially protected cerulein-injured ARJ cells by increasing cell viability,
reducing LDH release, and promoting apoptosis. Rhein, naringin, and honokiol were the main absorbed components from DCQD
in cerulein-induced pancreatitis. Moreover, rhein in combination with naringin and honokiol had synergistic eects in protecting
cerulein-injured ARJ cells and was better than the individual or the pairwise combination of the three components. Conclusions.
e ten eective components from DCQD may elicit similar protective eects as DCQD on cerulein-induced pancreatitis. e
principle of the formula compatibility of DCQD may be identied based on the eects of its absorbed components in cerulein-
injured ARJ cells.
1. Introduction
Dachengqi decoction (DCQD) is traditional Chinese herbal
decoctioncomposedofRadix et Rhizoma Rhei (Dahuang),
Cortex Magnoliae Ocinalis (Houpo), Fructus Aurantii
Immaturus (Zhishi), and Natrii Sulfas (Mangxiao). It has been
used to treat acute pancreatitis (AP) for more than three
decades in China []. Recent studies reported that DCQD
could promote the gastrointestinal motility and inhibit the
cytokine’s activity and the inammatory response in patients
with AP [, ]. According to the prescription compatibility
principle of traditional Chinese medicine (TCM) formu-
lations, Dahuang is the principal drug in DCQD, while
Mangxiao, Houpo, and Zhishi are assistant ingredients. Our
previous studies found that good prescription compatibility
in TCM could increase the level of rhein and aect its
pharmacokinetics [, ]. Unfortunately, the related optimal
prescription compatibility of the absorbed components from
DCQD in AP remains unclear.
AP is one disease largely depending on the balance
between apoptosis and necrosis in pancreatic acinar cells. e
induction of apoptosis might be one self-protective factor
protecting against acinar cell injury and reducing the severity
of pancreatitis because of the release of variety intracellular
zymogenwhichcouldreducetheinammatoryresponse
[, ]. It has been reported that the induction of pancreatic
Hindawi Publishing Corporation
Evidence-Based Complementary and Alternative Medicine
Volume 2016, Article ID 3198549, 8 pages
http://dx.doi.org/10.1155/2016/3198549
Evidence-Based Complementary and Alternative Medicine
acinar cell apoptosis by crambene protects mice against AP
by the induction of anti-inammatory pathways [], whereas
for necrosis, in which cell membrane integrity is lost in
association with the release of digestive enzymes, lactate
dehydrogenase (LDH) and inammatory mediators can lead
to local and systemic inammatory response and damage [,
–]. Furthermore, our previous studies found that DCQD
could inhibit local and systematic inammatory responses
and alleviate pancreatic damage by regulating the necrosis-
apoptosis switch of the pancreatic cells in AP []. e ten
bioactive components of DCQD, namely, rhein, emodin,
aloe-emodin, chrysophanol, rheochrysidin, naringin, narin-
genin, hesperidin, honokiol, and magnolol, were detected
in the serum of rats and dogs [, ]. However, it is
unclear whether these individual components or the related
combination has the similar eects to DCQD in the treatment
of AP. is study investigated the protective eect of the
individual component andthe related combinationby testing
cell viability, lactate dehydrogenase (LDH) release, and levels
of apoptosis and necrosis in cerulein-injured pancreatic
ARJ cells, to identify the herbal formula compatibility law
of DCQD based on the eects of its absorbed components on
cerulein-injured ARJ cells.
2. Materials and Methods
2.1. Materials. Rhein, emodin, aloe-emodin, chrysophanol,
rheochrysidin, naringin, naringenin, hesperidin, honokiol,
and magnolol were purchased from the Sichuan Academy
of Chinese Medicine Sciences (Chengdu, China). eir puri-
ties were higher than %. Spray-dried Dahuang, Houpo,
Zhishi, and Natrii Sulphas powders were purchased from
Chengdu Green Herbal Pharmaceutical Co. Ltd. (Chengdu,
China). e mean contents of the components from DCQD
detected three times in our previous study were as follows:
rhein, . mg/g; emodin, . mg/g; aloe-emodin, . mg/g;
chrysophanol . mg/g, rheochrysidin, . mg/g; naringin,
. mg/g; naringenin . mg/g; hesperidin, . mg/g; hon-
okiol, . mg/g; magnolol, . mg/g []. e peak con-
centrations of these ten components in serum when rats
were administered DCQD with g/Kg⋅BW, as reported
in the previous studies, are as follows: rhein, . ng/
mL; emodin, . ng/mL; chrysophanol, . ng/mL; rhe-
ochrysidin, . ng/mL; aloe-emodin, . ng/mL; mag-
nolol, . ng/mL; honokiol, . ng/mL; naringin, . ng/
mL; hesperidin, . ng/mL; and naringenin, .ng/mL
[, ]. e stock solutions of these components prepared
at the reported peak serum concentrations were diluted
/, /, /, /, and / in distilled water. Spray-dried
powders were mixed with an equal amount of distilled
water and reconstituted in dimethylsulfoxide (DMSO) to
prepare a mg/mL stock solution and kept in −∘C.
e nal DMSO concentration was less than .% when
the components were added to the cell culture media. e
dosage required was calculated and samples were diluted
and quantitatively analyzed by high-performance liquid
chromatography (HPLC). Fetal bovine serum (FBS) was
obtained from HyClone (Logan, UT). DMSO, cerulein, FK
medium, and ,-dichlorouorescin diacetate (DCFH-DA)
were obtained from Sigma (St. Louis, MO, USA).
2.2. Methods
2.2.1. Cell Culture. e rat pancreatic acinar ARJ cells
(ATCC, Rockville, MD, USA) were cultured in FK medium
containing % FBS, U/mL penicillin, and 𝜇g/mL
streptomycin under standard conditions (∘Cand%CO
2).
All experiments were performed h aer cells were seeded
into culture plates. ARJ cells were treated for min with
or without the selected dosage of the components and then
coincubated with cerulein ( nM) for a further h.
2.2.2. Cell Viability Assay. Cell survival was assessed using
the WST viability assay kit containing WST- (-(-methoxy-
-nitrophenyl)--(-nitrophenyl)--( ,-disulfophenyl)-H-tet-
razolium, monosodium salt) according to the manufacturer’s
protocol (Roche, Basel, Switzerland). ARJ cells were
plated into -well plates at a density of ×4cells/well.
Aer incubation for h, cells were pretreated with various
concentrations of the components and coincubated with
cerulein for a further h. Aer cerulein is added, cell
viability was detected by the kit at h, h, h, h, and h.
WST- solution (. mg/mL) was added to each well and
cells were incubated at % CO2and ∘Cforh.Cellviability
was determined by the dierent absorbance at wavelengths
of nm. e relative cell viability rate was calculated using
the following formula: cell viability rate (%) = % ×mean
absorbanceofcellsinthetreatedgroup/meanabsorbanceof
cells in control group.
2.2.3. LDH Assay. Necrotic cell death was assessed by the
releaseofLDHfromthecytosolofdamagedcellsintothe
supernatant using the LDH Cytotoxicity Detection Kit (Nan-
jing Jiancheng Bioengineering Institute, Nanjing, China) at
various time points (– h) according to the manufacturer’s
instructions. e percentage of total cellular LDH released
was determined using the following equation: LDH release
(%) = total extracellular LDH activity at the given time point
×/total LDH activity at the given time point.
2.2.4. Apoptosis Assay. Cells were stained with the Annexin
V-FITC Apoptosis Detection Kit (Nanjing Kaiji, Nanjing,
China) according to the manufacturer’s instructions to detect
early apoptotic cells (annexin V+and propidium iodide
(PI)−) and necrotic or late apoptotic cells (annexin V+and
PI+) by ow cytometry. Briey, ARJ cells were treated
with or without various components for min and then
stimulated with cerulein ( nM) for h. Cells were collected
and resuspended in culture medium at a density of ×
6cells/mL and stained with 𝜇L of annexin V-FITC and
𝜇L PI prepared in 𝜇L binding buer ( mM HEPES,
pH ., mM NaOH, and . mM CaCl2) according to the
manufacturer’s instructions for min at room temperature
in the dark. Cells were analyzed by ow cytometry (FACScan,
Becton Dickinson, USA).
2.2.5. Statistical Analysis. Statistical analyses were performed
using the PEMS. statistics program. All data represented
at least three independent experiments and were expressed
as the mean ±standard deviation (mean ±SD). One-way
Evidence-Based Complementary and Alternative Medicine
T : e eects of the absorbed components from DCQD
individually on cerulein-induced ARJ cell death and LDH release.
Component Cell viability (%) LDH release (%)
Rhein . ±. . ±.
Emodin . ±. . ±.∗
Chrysophanol . ±. . ±.∗
Rheochrysidin . ±. . ±.∗
Aloe-emodin . ±.∗. ±.∗
Naringin . ±. . ±.
Hesperidin . ±.X. ±.X
Naringenin . ±.X. ±.X
Honokiol . ±. . ±.
Magnolol . ±.. ±.
Cerulein alone . ±. . ±.
Normal . ±.∗. ±.∗
Rhein, emodin, chrysophanol, rheochrysidin and aloe-emodin are from
Dahuang, naringenin, naringin and hesperidin are from Zhishi, and mag-
nolol and honokiol are from Houpo. LDH = lactate dehydrogenase. Cells
were pretreated with the ten components with the peak concentrations for
min and then coincubated with nM cerulein for h. Aer cerulein
added, cell viability examined by WST- assay. Necrotic cell death was
assessed by the release of LDH from the cytosol of damaged cells into the
supernatant using the LDH Cytotoxicity Detection Kit. e results are mean
±SD. ∗𝑝 < 0.05 versus rhein-treated group, X𝑝 < 0.05 versus naringin-
treated group; 𝑝 < 0.05 versus honokiol-treated group.
repeated-measures ANOVA (followed by multiple pairwise
comparisons using the Student-Newman-Keuls procedure)
was used to analyze dierences between experimental and
control groups. 𝑝 < 0.05 was regarded as statistically
signicant.
3. Results
3.1. Treatment with the Ten Components Individually from
DCQD Increased Cell Viability in Cerulein-Injured AR42J
Cells. To examine the eects of various components on cell
viability, pancreatic acinar ARJ cells were pretreated with
six concentrations (undiluted or diluted /, /, /, /, or
/) of each component and cerulein for h. Cell viability
was markedly decreased in cerulein-treatment group. Each
component had a protective eect on cerulein-induced cell
death in a concentration-dependent manner, and the most
proper treatment concentration was undiluted one, the peak
concentration (Figure ).
3.2. Treatment with the Ten Components from DCQD Individ-
ually Reduced Cell Death and LDH Release in Cerulein-Injured
AR42J Cells (Table 1). e eects of various components of
DCQD on the viability of ARJ cells and their LDH release
upon cerulein treatment were assessed. Viability was highest
among rhein-treated cells than those treated with emodin,
aloe-emodin, chrysophanol, or rheochrysidin from Dahuang
(Table ). LDH release was lowest among rhein-treated cells
than those treated with any other component from Dahuang
(𝑝 < 0.05). In comparison with cells treated with hesperidin
or naringenin from Zhishi, naringin-treated cells exhibited
ahigherviability(𝑝 < 0.05)andlowerLDHrelease(𝑝<
T : e eects of rhein, honokiol, and naringin individually
or in combination on cerulein-induced ARJ cell death and LDH
release.
Component Cell viability (%) LDH release (%)
R.±.∗X. ±.∗X
H.±.∗X. ±.∗X
N . ±.∗X. ±.∗X
R plus H . ±.∗X. ±.∗X
RplusN .±.∗X. ±.∗X
NplusH .±.∗X. ±.∗X
R plus H plus N . ±.∗X. ±.∗X
Ten components . ±.∗X. ±.∗X
DCQD . ±.∗ . ±.∗
Cerulein alone . ±. . ±.
Normal . ±.∗. ±.∗
R = rhein, H = honokiol, N = naringin, DCQD = Dachengqi decoction, and
LDH = lactate dehydrogenase. Cells were pretreated with the components
with the peak concentrations for min and then coincubated with nM
cerulein for h. Aer cerulein is added, cell viability is examined by WST-
assay. Necrotic cell death was assessed by the release of LDH from the cytosol
of damaged cells into the supernatant using the LDH Cytotoxicity Detection
Kit. e results are mean ±SD. ∗𝑝 < 0.05 versus cerulein alone-treated
group, 𝑝 < 0.05 versus rhein-treated group, and X𝑝 < 0.05 versus DCQD-
treated group.
0.05). Cell viability was higher and LDH release was lower
among honokiol-treated cells than that among magnolol-
treated cells (𝑝 < 0.05). Rhein, naringin, and honokiol may
bethemainabsorbedcomponentsofDahuang,Zhishi,and
Houpo, respectively.
3.3. Treatment with Combination Components from DCQD
Reduced Cell Death and LDH Release in Cerulein-Induced
AR42J Cells. According to the aforementioned results, rhein
is a major absorbed component from Dahuang and pro-
tects against cerulein-injured ARJ cells and reduces LDH
release. Similarly, naringin and honokiol are considered to
be the major absorbed components of Zhishi and Houpo,
respectively. What is more, the most eective concentration
was the peak concentration according to our rst result;
therefore, we explored whether treatment with combinations
of the three major components in peak concentrations or with
the mixture of the ten components induced similar eects
to DCQD. e levels of cerulein-induced ARJ cell death
and LDH release were measured aer pretreatment with the
various combinations of components (Table ). Compared
to untreated cells, the cell viability was signicantly reduced
among the cells treated with cerulein. However, this decrease
in the cell viability was signicantly prevented among the
cells pretreated with rhein, honokiol, naringin, rhein plus
honokiol, rhein plus naringin, naringin plus honokiol, rhein
plus honokiol and naringin, the mixture of the ten com-
ponents, or DCQD. e cell viability was higher among
rhein-treated cells than honokiol plus naringin-treated cells.
e cell viability increased signicantly when the cells were
treated with rhein plus honokiol, rhein plus honokiol and
naringin,orthemixtureofthetencomponents.However,the
cell viability was higher among the cells treated with DCQD
Evidence-Based Complementary and Alternative Medicine
1/4
1/8
1/16
1/32
Normal
Cerulein
1/2
100
90
80
70
60
50
40
30
20
10
0
Rhein
0 2 4 6 8 10 12 14
(h)
16 18 20 22 24
Cell viability (%)
=365.67 ng/mL1∗time
Emodin
100
90
80
70
60
50
40
30
20
10
0
02468101214
(h)
16 18 20 22 24
Cell viability (%)
=3.62 ng/mL1∗time
Chrysophanol
100
90
80
70
60
50
40
30
20
10
0
02468101214
(h)
16 18 20 22 24
Cell viability (%)
=36.33 ng/mL1∗time
Magnolol
100
90
80
70
60
50
40
30
20
10
0
02468101214
(h)
16 18 20 22 24
Cell viability (%)
=1.08 ng/mL1∗time
Aloe-emodin
100
90
80
70
60
50
40
30
20
10
0
0 2 4 6 8 10 12 14
(h)
16 18 20 22 24
Cell viability (%)
=10.07 ng/mL1∗time
Rheochrysidin
100
90
80
70
60
50
40
30
20
10
0
02468101214
(h)
16 18 20 22 24
Cell viability (%)
=1.83 ng/mL1∗time
Hesperidin
100
90
80
70
60
50
40
30
20
10
0
0 2 4 6 8 10 12 14
(h)
16 18 20 22 24
Cell viability (%)
=40.95 ng/mL1∗time
Naringenin
100
90
80
70
60
50
40
30
20
10
0
0 2 4 6 8 10 12 14
(h)
16 18 20 22 24
Cell viability (%)
=22.67 ng/mL
1∗time
Naring in
100
90
80
70
60
50
40
30
20
10
0
02468101214
(h)
16 18 20 22 24
Cell viability (%)
=42.83 ng/mL
1∗time
Honokiol
100
90
80
70
60
50
40
30
20
10
0
0 2 4 6 8 10 12 14
(h)
16 18 20 22 24
Cell viability (%)
=9.07 ng/mL1∗time
1∗time
F : e eects of the ten absorbed components from DCQD on the cerulein-induced necrosis of ARJ cells. Cells were pretreated
with various concentrations (1∗time, /, /, /, /, or /) of each component for min and then coincubated with nM cerulein for
h. Aer cerulein is added, cell viability is examined by WST- assay at h, h, h, h, and h. 1∗time concentration means the peak
concentration of the components in serum detected by our previous study.
Evidence-Based Complementary and Alternative Medicine
than among those treated with any of the components, either
individually or in combination, including those treated with
all ten components. LDH release was relatively low in the
untreated cells and was signicantly increased by cerulein
treatment. Pretreatment with rhein, honokiol, naringin, rhein
plus honokiol, rhein plus naringin, naringin plus honokiol,
rhein plus honokiol and naringin, the mixture of the ten
components, or DCQD signicantly reduced LDH release.
LDH release was lower among the cells treated with DCQD
than among those treated with any of the components,
either individually or in combination. LDH release was lower
among the cells treated with rhein than among those treated
with honokiol and/or naringin. All these results showed
that the components, either individually or in combination,
including the mixture of ten components demonstrated some
ecacy of the prescription of DCQD.
3.4. Treatment Individually or in Combination with the Com-
ponentsofRhein,Honokiol,andNaringinfromDCQDShowed
a Synergistic Eect on the Apoptosis-Necrosis Cellular Switch
in Cerulein-Induced AR42J Cells. Our previous study found
that DCQD could regulate the apoptosis-necrosis switch of
pancreaticacinarcellsinratswithAPorinisolatedcells.
Annexin V/PI staining was performed to assess whether the
level of apoptosis diered among samples treated with the
three major bioactive components of DCQD, individually
or in combination. Annexin V−/PI−cells were regarded as
healthy, annexin V+/PI−cells were regarded as early apop-
totic, and annexin V+/PI+cells were regarded as necrotic or
late apoptotic (Figure ). Flow cytometry analyses suggested
an extremely low level of cell death in untreated samples,
and this was markedly increased following treatment with
cerulein for h. e individual treatment, the pairwise
combination, or all the three components of rhein, honokiol,
andnaringincouldincreasetherateofapoptosisinARJ
cells, but the percentage of apoptotic cells treated with all
the three components of rhein, honokiol, and naringin was
signicantly highest among all the six treatment groups (𝑝<
0.05) (Table ), which showed a synergistic eect on the
apoptosis-necrosis cellular switch in cerulein-induced ARJ
cells.
4. Discussion
is study identied the protective eect of individual com-
ponent and related combination of the components from
DCQD in dose-dependent and time-dependent manner on
cerulein-induced ARJ cells. Among the ten components,
pretreatment with rhein of the peak serum concentration in
cerulein-induced ARJ cells showed the strongest protective
eect among the components from Dahuang, such as cell
viability and LDH release, on injured ARJ cells. Similarly,
naringin and honokiol showed similar protective eect in
injured ARJ cells. Rhein, naringin, and honokiol may be
the major eect components from DCQD in treatment of AP
in vitro. All these results showed that the components, either
individually or in combination, have some ecacy of the
T : e proapoptotic eects of rhein, honokiol, and naringin
from DCQD on cerulein-induced ARJ cells.
Component Apoptotic cells (%)
R.±.#X
H.±.#X
N . ±.#X
RplusH .±.#X
R plus N . ±.#X
N plus H . ±.#X
R plus H plus N . ±.#X
Ten components . ±.#X
DCQD . ±.#
Cerulein alone . ±.∗
Normal . ±.#
R = rhein, H = honokiol, N = naringin, and DCQD = Dachengqi decoction.
Ten components = the mixture of the ten components from DCQD. ARJ
cells were pretreated with or without rhein, honokiol, and naringin with
the peak concentrations individually or in combination for min and
then stimulated with cerulein ( nM) for h. Cells were stained with the
Annexin V-FITC Apoptosis Detection Kit and analyzed by ow cytometry.
e results are mean ±SD. ∗𝑝 < 0.05 versus normal group; 𝑝 < 0.05 versus
rhein-treated group; X𝑝 < 0.05 versus DCQD-treated group; #𝑝 < 0.05
versus cerulein alone-treated group.
prescription of DCQD. It is similar to the study of Guan-Xin-
Er-Hao formula [] which indicated that the combination of
three absorbed bioactive components (ferulic acid, tanshinol,
and hydroxysaor yellow A) is similar to its formula in
reducing infarct size of acute myocardial infarction (AMI) in
rats. e study of Ju-Zhi-Jiang-Tang (JZJT) [] showed that
the two major active constituents (nobiletin and tangeretin)
can signicantly exert anti-inammatory eects representing
the ecacy of the formula. Another research [] suggested
that claycosin and formononetin from Yu-Ping-Feng-San
(YPFS) can reduce allergic inammation similar to the
eectofYPFSinvivoandinvitro.Similarly,themajor
eective components of Shaoyao-Gancao decoction [], Bu-
Shen-Yi-Qi Fang [, ], Zhi-zi-chi decoction [], Yin-
Chen-Hao-Tang [], and so forth represent part ecacy
of formula. erefore, many major absorbed components of
herbs individually or in combination partly have ecacy of
formula.
Interestingly, combining rhein with honokiol, but not
with naringin, had an additive protective eect. Pretreat-
ment with naringin and honokiol elicited similar eects
on proapoptosis as treatment with rhein. Pretreatment with
the three components showed more positive eects than
treatment with each of them alone. With respect to the
regulation of the necrosis-apoptosis switch, treatment with
rhein, naringin, honokiol, rhein plus naringin, rhein plus
honokiol, naringin plus honokiol, rhein plus naringin plus
honokiol, all ten components, or whole DCQD promoted
injuredcellapoptosis.epercentageofapoptoticcellswas
higher among cells treated with rhein plus naringin plus
honokiol and all ten components than those treated with
rhein alone (𝑝 < 0.05) (Table ). Moreover, the percentage of
Evidence-Based Complementary and Alternative Medicine
0.1 1000
0.1
1000
(a) Rhein
0.1 1000
0.1
1000
(b) Honokiol
0.1 1000
0.1
1000
(c) Naringin
0.1 1000
0.1
1000
(d) Rhein + honokiol
0.1 1000
0.1
1000
(e) Rhein + naringin
0.1 1000
0.1
1000
(f) Honokiol + naringin
0.1 1000
0.1
1000
(g) Rhein + honokiol + naringin
0.1 1000
0.1
1000
(h) Ten compounds combinations
0.1 1000
0.1
1000
(i) DCQD
0.1 1000
0.1
1000
(j) Cerulein
0.1 1000
0.1
1000
(k) Normal
F : e synergistic eects of rhein, honokiol, and naringin from DCQD on the apoptosis-necrosis cellular switch in cerulein-induced
ARJ cells. Cells were pretreated with or without the three major components with the peak concentrations individually or in combination
for min and then coincubated with nM cerulein for h. Annexin V/PI staining was performed and ow cytometry analyses were
used. Each panel is divided into four regions: viable cells (annexin V−/propidium iodide (PI)−) are located in the lower le quadrant, early
apoptotic cells (annexin V+/PI−) in the lower right quadrant, late apoptotic and necrotic cells (annexin V+/PI+) in the upper right quadrant,
and primary necrotic cells (annexin V−/PI+) in the upper le quadrant.
Evidence-Based Complementary and Alternative Medicine
apoptotic cells was highest among cells treated with DCQD
(𝑝 < 0.05). is identied the herbal formula compatibility
based on the synergistic eects of rhein, honokiol, and
naringin from DCQD in vitro study on the pancreatic ARJ
cells. Many researches have conrmed the compatibility
principle of formula or herbs via the combination of the
major eective components in vivo or in vitro, especially the
additive or synergistic eects of the absorbed components.
e study of Sini decoction [] proved that the major active
ingredients (the total alkaloids, total gingerols, total avones,
andtotalsaponins)weremoreeectivethanformulasformed
by any one or two of the three individual components. In
another study [], amygdalin and hydroxysaor yellow A,
main components of Taoren-Honghua (TH) herb pair, are
responsible for the main curative eects of TH and usually
have synergetic eects, such as decreasing plasma viscosity
and platelet aggregation percentage. What is more, total
coumarins and volatile oil, as the two main components
of Radix Angelicae dahuricae, can improve the intestinal
absorption of baicalin extracted from Scutellaria baicalensis
Georgi andhavesynergisticactionintheenhancedabsorp-
tion of baicalin, where Angelicae dahuricae and Scutellaria
baicalensis Georgi is one herb pair, which has claried the
compatibility principles of herb pairs []. In our study, rhein,
naringin, and honokiol had synergistic eects, and naringin
plus honokiol had an additive eect on rhein in proapoptosis,
which proved the compatibility principle of herbs.
In this study, treatment with each of the ten absorbed
components of DCQD could elicit partial eects as DCQD
treatment on cerulein-induced pancreatitis. Rhein, naringin,
and honokiol may be the major eect components of
DCQD in treatment of AP in vitro. Moreover, treatment
with combinations of these three components elicited better
synergistic eects on cerulein-induced pancreatic acinar cells
than individual components treat ment, which help to identify
the herbal formula compatibility law of DCQD based on
the eects of its absorbed components on cerulean-injured
ARJ cells. Further studies are needed to determine the
optimal ratio of rhein, naringin, and honokiol for protection
against acinar cell death and to elucidate the underlying
mechanism.
Abbreviations
DCQD: Dachengqi decoction
AP: Acute pancreatitis
LDH: Lactate dehydrogenase
DMSO: Dimethylsulfoxide
HPLC: High-performance liquid chromatography
FBS: Fetal bovine serum
DCFH-DA: ,-Dichlorouorescin diacetate
AMI: Acute myocardial infarction.
Conflict of Interests
e authors conrm that this paper content has no conict of
interests.
Authors’ Contribution
Yumei Zhang and Lin Zhu contributed equally to this paper.
Wenfu Tang designed the study. Yumei Zhang, Jia Wang,
XianlinZhao,LinZhu,JuanLi,andHuiGuowereresponsible
fortheacquisitionofdata.YumeiZhangandJiaWangwere
responsible for the analysis and interpretation of data. Yumei
Zhang and Lin Zhu were responsible for draing of the
paper. Wenfu Tang was responsible for the critical revision
of the paper. Wenfu Tang obtained funding and studied
supervision.
Acknowledgment
e authors thank the support of the National Natural Sci-
ence Foundation of China (no. and no. ).
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