A Study of the Effect of Shiunko, a Traditional

Chinese Herbal Medicine, on Fibroblasts and Its

Implication on Wound Healing Processes

Kin-Fu Chak,1,2,* Chia-Yen Hsiao,1 and Ting-Yu Chen1

1
Institute of Biochemistry and Molecular Biology, National Yang Ming University, Taipei, Taiwan.
2
Institute of Biomedical Sciences, Mackay Medical College, New Taipei City, Taiwan.

Significance: In China, traditional Chinese medicine (TCM) has been used for
thousands of years for various acute and chronic wound care. Thus, there is a
growing need to explore the possible benefits of TCM on wound healing.
Recent Advances: Nowadays, in China and some Asian countries including
Korea, Japan, and Singapore, Chinese herbal therapy is used as an alternative
treatment in wound care. Therefore, exploration of the possible benefits of
TCM on wound healing is necessary.
Critical Issues: Development of TCM is based on the concept of Yin (negative
phenomenon of nature) and Yang (positive phenomenon of nature). These
Kin-Fu Chak, PhD opposing and complementary natural phenomena of the universe restore the
Submitted for publication November 26,
normal physiological functions, consequently curing diseases and restoring
2012. health of a patient.
*Correspondence: Institute of Biomedical Sci- Future Directions: Due to lack of evidence-based research, TCM treatments
ences, Mackay Medical College, New Taipei City
252, Taiwan (e-mail: [email protected]).
are not widely accepted in the western world. Using state-of-the-art technol-
ogy such as proteomics, bioinformatics, and biomolecular techniques, research
studies may lead to more effective remedies for wound care in the future.

Abbreviations
and Acronyms
ARPC5 = actin-related protein SCOPE TRANSLATIONAL RELEVANCE
2/3 complex subunit 5 The balance of two opposite A large number of TCM literatures
AS = Angelica sinensis natural phenomena, Yin (negative) have documented the formulation
ATPD = ATP synthase subunit and Yang (positive) of the body, gov- of herbal medicines for various
delta erns everything in the world, and are wound treatments. With the aim of
CAPNS1 = calpain small the sources of growth, development, evidence-based research, this review
subunit 1 and variation of all organisms.1,2 The may explore a better formulation of
COF2 = cofilin-2 concept of traditional Chinese medi- TCM for various wound healing
CSU = chronic skin ulcer cine (TCM) literatures stated that treatments of patients.
FA = ferulic acid poor circulation, stagnation, and poor
GLRX-3 = glutaredoxin 3 healing are mainly due to the defi-
ciency of Yang, whereas overheating CLINICAL RELEVANCE
GSTP1 = glutathione S-
transferase P (a terminology of TCM for inflamma- Using the fibroblast as a cell model
HSPB1 = heat shock 27 kDa
tion and some related pathological system, we have adapted proteomics
protein 1 symptoms) or having an excess of scar and bioinformatics approaches to
IF5A-1 = eukaryotic translation
tissues are always associated with the evaluate the potential efficacy of
initiation factor 5A-1 deficiency of Yin. The balance between Shiunko, an herbal Chinese com-
Yin and Yang is ideal for good health, pound drug, on wound healing pro-
(continued) and it is essential for promoting wound cesses. Using proteomic analysis, we
healing in a complementary way. have identified some specific protein

448 j ADVANCES IN WOUND CARE, VOLUME 2, NUMBER 8

Copyright ª 2013 by Mary Ann Liebert, Inc. DOI: 10.1089/wound.2012.0368
 TRADITIONAL CHINESE MEDICINE IN WOUND CARE 449

factors associated with the function drug has the effects of arresting
of wound healing process. Thus, this bleeding by resolving blood stasis Abbreviations
evidence-based platform may explore and promoting blood circulation to and Acronyms (continued)
the beneficial molecular basis of relieve swelling and pain. It is often
LDHB = lactate dehydrogenase
TCM for wound healing as a whole. prescribed to treat traumatism, pain B
caused by ecchymoma and carbuncle,
LR = Lithospermi Radix
swelling, and other external skin
BACKGROUND PGK1 = phosphoglycerate
diseases. Other herbal hemostatics3
The Chinesis Materia medica kinase 1
including field thistle [Herba Cepha-
The aim of combining different PHB = probibitin
lanoploris; Cephalanoplos segetum
herbal drugs to treat diseases is PKM2 = pyruvate kinase
(Bunge) Kitam., or Cephanoplos se-
to rectify the hyperactivity or hy- isozyme type M2
tosum (Willd.) Kitarn.], sanguisorba
poactivity of Yin or Yang, resulting in PRDX = peroxiredoxin
root (Radix Sanguisorbae; Sangui-
restoration of normal physiological ROS = reactive oxygen species
sorba officinalis), hyacinth bletilla
functions of the patient, which con-
[Rhizoma Bletillae; Bletilla striate SH3BGRL = SH3 domain-
sequently cures diseases and re- binding glutamic acid–rich
(Thunb.) Reichb.], and cat-tail pollen
stores health.3 In practice, the herbal protein
(Pollen Typhae; Typha angustifolia L.
formulas normally consist of four SODC = superoxide dismutase
or Typha orientalis Presl) are useful
categories of herbs to rectify the bal- C
for treating various skin wounds.
ance of Yin and Yang: SPEE = spermidine synthase
 Ministerial herbs (also known Drugs for promoting blood circulation STMN1 = stathmin 1
as Chief herbs) address the and removing blood stasis. These TCM = traditional Chinese
principal pattern of the disease; drugs are effective for various syn- medicine
dromes due to obstruction of blood TCTP = translationally
 Deputy herbs assist the minis-
circulation and retention of blood controlled tumor protein
terial herb;
stasis in traumatic injuries, arthral- TGFb = transforming growth
 Assistant herbs are designed to gia, sores, carbuncles, and other factor beta
reduce the side effects of the suppurative skin infections.3 The TPIS = triosephosphate-
first two classes of herbs; and commonly used herbal drugs in isomerase
 Envoy herbs detoxify and direct this category are red sage root VIME = vimentin
the therapy to a particular part (Radix Salviae Miltiorrhizae; Salvia
of the body. miltiorrhiza), motherwort (Herba
Leonuri; Leonurus cardiaca), and
Principle of TCM for wound care safflower (Flos Carthami; Cartha-
Drugs for hemostasis (to stop in- mus tinctorius L.).
ternal and external bleeding) and for
promoting blood circulation and re- Other drugs for assistance of
moving blood stasis are regarded as wound care. For maintaining the
potent drugs of TCM for wound care.3 physiological harmony of patients,
properties of other drugs are usually
Hemostatics. These herbal drugs combined with the ministerial drugs
have the effects of stopping bleeding to optimize the efficacy of the drugs
by removing heat (inflammation and for wound care.3 For example, musk
related pathological symptoms) from (Moschus) and borneol (Borneolum)
the blood, by astringency, or by re- with resuscitative properties are al-
solving blood stasis. They are indi- ways formulated with other drugs in
cated for the treatment of various the TCM wound healing medicines.
kinds of hemorrhage and traumatic Moreover, drugs tonifying the defi-
bleeding. cient Qi (the most basic element for
Notoginseng (Radix Notoginseng), the composition and maintenance of
for example, is the root of Panax no- the vital activities of the human
toginseng, which is the most power- body) and rectifying Yin and Yang of
ful herbal hemostatics of TCM.4 This the body are often accompanied with
450 CHAK ET AL.

other drugs for wound care, such as chuanxiong anti-bacterial and anti-inflammatory effects has
rhizome (Rhizoma Ligustici Chuanxiong; Ligusti- been proven.10
cum chuanxiong Hort.), astragalus root (Radix As-
tragali seu Hedysart; Astragalus propinquus), red Other commonly used traditional Chinese herbal
sage root (Radix Salviae Miltiorrhizae), motherwort medicine for wound care. Various ancient and
(Herba Leonuri), Chinese yam (Rhizoma Dioscor- modern TCM literature have documented the use
eae; Dioscorea opposita), licorice (Radix Glycyr- of herbal formulas in wound care,11 such as Wuwei
rhizae; Glycyrrhiza glabra), Chinese angelica root Xiaodu Yin (clears heat, detoxifies, cools blood, and
(Radix Angelicae Sinensis; Angelica sinensis [AS]), reduces swelling), Huanglian Jiedu Tang (detoxi-
fleeceflower root (Radix Polygoni Multiflori; Fallo- fication decoction with huanglian), Yanghe Tang
pia baldschuanica), and achyranthes and cyathula (decoction for Yang harmonization, tonifies blood,
root (Radix Achyranthis Bidentatae et Radix Cy- expels cold, and unblocks stagnation), Tuoli Xiaodu
athulae; Achyranthes bidentata Bl. and Cyathula San (power for defensive Qi enrichment and toxi-
officinalis Kuan). cant removal), and Shengji Yuhong Gao (red oint-
ment for granulation promotion).
The TCM strategy for wound care
Three golden rules of TCM5 for wound care, es- Current clinical application of TCM
pecially chronic skin ulcers (CSU), have long been for wound care
proposed: (1) keeping the right amount of pus on In China, CSU patients account for 1.5%–3% of
the surface of the ulcer to stimulate the growth of the total hospitalized patients.12 Traditionally,
granulation, (2) removing necrotic tissues to stim- TCM remedy procedure for CSU is divided into the
ulate the growth of new skin, and (3) inhibition of following three steps.13
inflammation to promote skin wound recovery. In
practice, this strategy of treatment not only facili- Clearance of heat at the early stage of CSU.
tates wound healing but also effectively decreases The combination of the following herbal drugs is
scarring. commonly used for this purpose: Lonicera japonica
Thunb., Viola chinensis, Atractylodes lancea, Poria
cocos, Semen coicis, Stephania tetrandra S. Moore,
Yunnan Paiyao. Yunnan Paiyao (the white
AS, and Platycodi.
herbal medicine from Yunnan province of China)6
is the most famous of the patent remedies in China Promoting blood circulation to dissipate blood
for its ability to stop bleeding, for closure of wounds stasis for the treatment at the middle stage of
and decrease in scarring, and for the encourage- CSU. The combination of the following herbal
ment of flesh regeneration. drugs is commonly used for this purpose: Cartha-
Yunnan Paiyao is formulated by various herbal mus tinctorius, AS, Aucklandiae Radix, Olibanum,
drugs with specific characters: Radix Notoginseng Comiphora Myrrha, Rhizoma Chuanxiong, Pher-
(40%), Ajuga Forrestii Diels (17%), Rhizoma Dios- etima, Glycyrrhizae Radix, Salviae Miltiorrhizae
coreae (13%), Rhizoma Dioscoreae Nipponicae Radix, Rehmanniae Radix, and Paeoniae Radix.
(10%), Herba Geranli & Herba Erodii (7.2%),
Dioscoreae Parviflora Ting (6%), Herba Inulae Supplements for deficiency to promote granulation
Cappae (5%), Borneol (1.5%), and Musk (0.3%). at later stage of CSU. The combination of the
Recent research demonstrated that either before following herbal drugs is commonly used for this
or after the surgery, administration of Yunnan purpose: Astragalis Raw Radix, Pseudostellariae
Paiyao can significantly reduce bleeding and Radix, Angelicae Sinensis Radix, Salviae Miltior-
swelling of the wound.7 rhizae Radix, Semen Persicae, Carthami Flos,
Rhizoma Chuanxiong, Rhizoma Atractylodis Mac-
Shiunko. Shiunko8 is commonly used in China rocephalae, Poria Cocos, Cyathulae Radix, and
and Japan for treating wounded skin caused by Glycyrrhizae Radix.
cuts, abrasions, frost, or burn.9 Shiunko mainly
consists of sesame oil (500 mL), Lithospermi Radix
(LR; Lithospermum erythrorhizon Sieb. et Zucc.) PARADIGM OF EVIDENCE-BASED RESEARCH
(10 g), A. sinensis (10 g), lard (3 g), and beeswax OF TCM ON WOUND HEALING PROCESS
(45 g). The therapeutic effect of shiunko on granu- Proteomics is a powerful tool and has been widely
lation tissue formation, speeding wound healing, used to elucidate protein profile changes in response
reepithelialization, and angiogenesis, in addition to to drug treatment and to identify disease-relevant
 TRADITIONAL CHINESE MEDICINE IN WOUND CARE 451

biomarkers.14 In this work, proteomics and bioin-

formatics were employed to study the mechanism of
wound healing process of a TCM wound healing
drug, shiunko.
Shiunko, a Chinese herbal compound,
for wound healing
AS is a basic component of many Chinese drugs
for wound healing.9 Ferulic acid (FA) is one of the
most abundant water-soluble ingredients in AS,
and has been reported to be the active component of
AS.15,16 LR (also called Zicao or Gromwell) is com-
monly used to treat skin disorders, cuts, and burns.
A recent study showed that LR has multiple ac-
tivities including antimicrobial activity,17 antiviral
activity,18 anti-inflammatory activity,19 and anti-
tumor activity.20 The most important components Figure 1. Viability of fibroblast cells in response to various treatments of
Shiunko. Fibroblasts and designated concentrations of Shiunko were dis-
in LR are derivatives of shikonin, such as deoxy- solved in 0.5% dimethyl sulfoxide (DMSO). 24 h after treatment with various
shikonin, acetylshikonin, and isobutylshikonin.21 concentrations of Shiunko, the viability of fibroblast cells were measured
To study the molecular basis of wound healing when subjected to WST-1 assay. Fibroblast cells in 0.5% DMSO was used
processes of this herbal medicine, Shiunko was as control, and the relative viability of fibroblast cells treated with various
concentrations of Shiunko was determined by comparing with the control,
prepared with 1:1 (w/w) of LR/AS mixture and ex- respectively. The Student’s t-test was used to evaluate the statistical sig-
tracted by 95% ethanol. nificance of the experiments, and the results were presented as mean – SD
(n = 3), where **p < 0.01.
Proteomic approach for the analysis of proteins
associated with the wound healing process. The
differentially expressed proteins of fibroblasts in-
duced by Shiunko were dominantly associated with AS (300 lg/mL)22 and LR (50 lg/mL)23 was only
many pathological functions (our unpublished 120% to 125%. Obviously, the combination of AS
data), including antioxidant activity (peroxiredoxin (50 lg/mL) and LR (50 lg/mL), designated Shiunko
[PRDX]2, PRDX4, superoxide dismutase C [SODC], 100, has a synergistic effect on cell proliferation
glutathione S-transferase P [GSTP1], glutaredoxin (Fig. 1). We found that the expression level of the
3 [GLRX-3], and SH3 domain-binding glutamic anti-apoptotic proteins, such as HSPB1, GSTP1,
acid–rich protein [SH3BGRL]); cell proliferation and TCTP, in cells induced by AS, LR, and Shiunko,
(pyruvate kinase isozyme type M2 [PKM2], sper- respectively, was similar. However, the expression
midine synthase [SPEE], probibitin [PHB], stath- level of proliferation proteins was very different.
min 1 [STMN1], and eukaryotic translation Our previous data indicated that AS (300 lg/mL)22
initiation factor 5A-1 [IF5A-1]); anti-apoptosis could only induce an increase in the expression
(GSTP1, heat shock 27 kDa protein 1 [HSPB1], and level of CAPNS1; LR (50 lg/mL)23 could induce an
translationally controlled tumor protein [TCTP]); increase in the expression levels of IF5A-1 and
collagen secretion (phosphoglycerate kinase 1 NME-1. Interestingly, proteomic analysis of the
[PGK1], lactate dehydrogenase B [LDHB], ATP drug effect on fibroblasts indicated that more pro-
synthase subunit delta [ATPD], and triosepho- liferation proteins were induced by Shiunko
sphate-isomerase [TPIS]); and cell motility (MARE- 100 lg/mL, including IF5A-1, NME-1, PKM2,
1, vimentin [VIME], calpain small subunit 1 SPEE, PHB, and STMN1(our unpublished data). It
[CAPNS1], actin-related protein 2/3 complex sub- was known that STMN1 is a key proliferation
unit 5 [ARPC5], and cofilin-2 [COF2]). protein,24,25 and it can be regarded as a typical
differentiation protein marker of the cell.26 It is
Differentially expressed proteins associated with worth noting that Shiunko could induce an in-
cell viability and proliferation. The relative prolif- crease in expression level of STMN1 by up to 8.5-
eration rate of fibroblast cells induced by the opti- fold (our unpublished data), indicating that
mal concentration of Shiunko 100 lg/mL STMN1 may play an important role in the prolif-
(containing 50 lg/mL and AS and 50 lg/mL LR) eration rate of fibroblasts. Further, it could not be
was 150% (Fig. 1). In comparison, our previous ruled out that a higher proliferation rate of fibro-
works indicated that the proliferation rate of fi- blasts induced by Shiunko may be also due to more
broblast induced by the optimal concentration of proliferation proteins induced by this treatment.
452 CHAK ET AL.

Differentially expressed proteins associated with Correlation between collagen secretion and ex-
antioxidant ability. With discrete drug treatments pression of transforming growth factor beta. The
of the fibroblast cell, expression profiles of proteins collagen secretion rate of fibroblasts treated with
associated with antioxidants are varied: AS either LR23 or Shiunko 100 (Fig. 4) was increased
(300 lg/mL)22—PRDXs and PARK-7; LR (50 lg/ by up to 125% to 130%, while the increase of col-
mL)23—PRDX-2, PRDX-4, GSTP-1, SODC, and lagen secretion rate of the AS-treated fibroblasts25
ADI-1; Shiunko (100 lg/mL)—PRDX-2, PRDX-4, was only 105%. Transforming growth factor beta
GSTP-1, SODC, GLRX-3, and SH3BRL (our un- (TGFb) controls the upregulation of collagen syn-
published data). The biochemical analysis indi- thesis in macrophages and fibroblasts.28 Coin-
cated that treatment of AS (50 lg/mL) caused the cidentally, production of TGFb was increased by
reduction of ROS by 8%,22 while both LR (50 lg/ up to 10-fold in cells treated with either LR23 or
mL)23 and Shiunko (100 lg/mL) (Fig. 2) signifi- Shiunko 100 lg/mL (Fig. 5), while there was only
cantly reduced the level of ROS by up to 15%. 1.5-fold increase in cells with AS treatment.22 It
is noteworthy that when fibroblast was treated
Differentially expressed proteins associated with with Shiunko 300 lg/mL, collagen secretion rate
cell mobility. Proteins involved in cell mobility, and TGFb expression level increased by up to
such as MARE-1, CAPNS-1, COF-2, and VIME 160% (Fig. 4) and 39.65-fold (Fig. 5), respectively.
were downregulated when cells were treated with Hence, this data strongly demonstrated that
AS,22 LR23, and Shiunko (our unpublished data). It Shiunko leads to increased production of TGFb
was known that ARPC5 is able to regulate cell and collagen.
mobility, probably by controlling microtubule
dynamics.27 In this work, we found that ARPC5 The wound healing analysis in vivo. The effect of
was only upregulated in cells treated with FA;22 in Shiunko on the wound healing speed was confirmed
contrast, this protein was downregulated in cells by an in vivo test. Two 8 mm biopsy punch wounds
treated with LR,23 and with Shiunko treatment were created on the flank or back of SD rats, and
(our unpublished data). Coincidently, with FA or
AS treatment, the rate of cell migration increased
by up to 160%,22 while cell migration rate reduced
to 40%–50% in the cells treated with either LR23 or
Shiunko (Fig. 3). Clearly, ARPC5 plays a critical
role in cell migration.

Figure 3. Mobility of fibroblast cells in response to various treatments of

Shiunko. Transwell migration assay was used to evaluate the mobility of
the cells treated with various concentrations of Shiunko. A total of 2 · 104
cells were seeded onto the upper Boyden chamber, and after 6 h, number
of cells that migrated down to the lower chamber of Transwell was
strained by 5% Giemsa in water, and the cells that remained on the upper
Boyden chamber were scraped with cotton swabs. The total number of
Figure 2. Detection of reactive oxygen species (ROS) contents in fibro- cells was counted using an inverted microscope. Migration rate of cells in
blast cells with various treatments of Shiunko. The contents of ROS of 0.1% DMSO was used as control, and the migration rate was defined as
fibroblast cells in 0.5% DMSO was used as control, the ROS ratio of cells 100%. The relative migration rate of cells with various treatments of
with various treatments of Shiunko was determined by comparing with the Shiunko was determined by comparing with the control. Student’s t-test
control. The Student’s t-test was used to evaluate the statistical signifi- was used to evaluate the statistical significance of the experiments, and
cance of the experiments, and the results are presented as mean – SD the results are presented as mean – SD (n = 3), where *p < 0.05 and
(n = 3), where **p < 0.01. **p < 0.01.
 TRADITIONAL CHINESE MEDICINE IN WOUND CARE 453

Figure 4. Detection of collagen secretion from fibroblast cells treated

with various concentrations of Shiunko. Fibroblast cells and designated
concentrations of Shiunko were dissolved in 0.5% DMSO. 24 h after drug
treatments, cells were harvested and subjected to Sircol soluble collagen
assay. Collagen secretion rate of cells in 0.5% DMSO was used as
control. The relative collagen secretions rate of cells with various Figure 6. In vivo wound healing speed assay. Two wound circles were
treatments was determined by comparing with control. Student’s t-test generated on the back of SD rats using an 8 mm biopsy punch. DMSO alone,
was used to evaluate the statistical significance of the experiments, and and three discrete concentrations of Shiunko in DMSO were used to treat the
the results are presented as mean – SD (n = 3), where *p < 0.05 and wounds, and the diameter of each wound was measured twice per day. The
**p < 0.01. wound healing rate was also recorded by digital camera at an interval of 4
days. To see this illustration in color, the reader is referred to the web version
of this article at www.liebertpub.com/wound

then the wounds were treated with three different

concentrations of Shiunko: 30, 100, and 300 lg/mL,
respectively. We observed that the contraction with Shiunko 30 lg/mL was not as good as the other
speed of the wound treated with Shiunko 100 and two treatments. These experimental results might
300 g/mL was similar (Fig. 6), as the opening of the be due to the fact that production of both collagen
wound contracted to near zero at day 11 of treat- (Fig. 4) and TGFb (Fig. 5) from fibroblasts treated
ment. It was noted that the wound healing speed with Shiunko 30 lg/mL was much lower than that of
the other two treatments.

DISCUSSION
Shiunko-treated fibroblasts can induce a whole
range of biochemical events involved in the wound
healing process, including cell proliferation and
anti-apoptosis, anti-oxidant activity, secretion of
collagen, and cell mobility (Fig. 7). Increase in cell
viability illustrated the synergistic effect of
Shiunko on the wound healing process, which is
achieved by the upregulation of proteins associated
with proliferation and anti-apoptosis of the treated
fibroblast cells. It is worth noting that Stathmin, a
differentiation marker, was highly induced by
Figure 5. Detection of transforming growth factor beta (TGFb) expression Shiunko (eightfold increase, our unpublished
of fibroblast cells in response to the treatment with Shiunko. Fibroblast cells data); possibly, presence of this protein could be a
and the designated concentrations of shiunko were dissolved in 0.5%
good sign of the wound healing process.
DMSO. 24 h after drug treatment, TGF-b expression was determined by
immunoblotting, and the quantity of TGF-b was confirmed by using Fujifilm In the early phase of the wound healing process,
MultiGauge software. Expression level of TGFb of cells in 0.5% DMSO was overexpression of reactive oxygen species (ROS)
used as control. The relative expression of cells with various drug treat- results in the formation of a secondary injury of
ments was compared with control. This experiment was repeated thrice,
the wound. We observed that Shiunko-treated fi-
and GAPDH was used as internal control.
broblasts can reduce the ROS concentration by up
454 CHAK ET AL.

to 15%, which may alleviate the risk of

TAKE-HOME MESSAGES
scarring after the wound heals. GSTP1
and PRDX are involved in the processes The philosophical concept of TCM for medical care
The balance of Yin and Yang eventually invigorates the vital activities of the
of antioxidation; they are commonly ex-
human body, Qi, for good health. To rectify the Qi of the patient, a combination
pressed in cells with AS, LR, and
of four categories of herbal drugs—ministerial (chief), deputy, assistant, and
Shiunko treatment. Interestingly, ex-
envoy herbs—should be prescribed.
pression of SODC (3.1-fold, our unpub-
lished data) is high only in cells with Current practice of TCM for wound care
Shiunko treatment. The function of super- TCM remedy procedure for CSU is divided into three steps, namely, clearing
heat, resolving stasis, and tonification methods to treat the syndromes at early,
oxide dismutase (SOD) is to catalyze the
middle, and later stages of the disease, respectively.
dismutation of the toxic superoxide anion
–
O2 to O2 and H2O2. In addition, it also has a Evidence-based research of TCM for wound care
29
powerful anti-inflammatory activity, and The evidence-based research indicated that Shiunko modulates a group of
can reverse fibrosis through reversion of proteins such as stathmin, SODC, TGFb, and ARPC5 and acts to improve healing
30 by doing so.
myofibroblasts back to fibroblasts.
Therefore, upregulation of SOD may reveal
a good sign of wound healing process.
Collagen is mainly secreted from fibroblasts, a treatment.22 ARPC5, a positive cell mobility regu-
component of skin tissue that can benefit all stages lator, is only upregulated in cells with AS and FA
of the wound healing process. TGFb is an upstream treatment,22 while expression of this protein is
regulator to control expression of collagen. We suppressed in cells with Shiunko treatment (our
found that expression of TGFb is correlated with unpublished data). In the Shiunko-treated fibro-
the secretion rate of collagen in fibroblasts with blasts (Figs. 4 and 5), increased collagen expression
Shiunko treatment. Hence, upregulation of TGFb was found to be concomitant with overexpression of
should be one of the important factors for wound TGFb. It is foreseeable that retention of a certain
healing process. amount of fibroblasts in the wound area during the
Mobility of fibroblast is restricted when the cell wound healing process is important. It is note-
is treated with Shiunko. In contrast, cell migration worthy that an increase of collagen production
increases by up to 150%–160% with AS and FA leads to downregulation of collagenase synthesis.31
Collagenase is a key enzyme for catalyzing the
extracellular matrix; therefore, downregulation of
collagenase may result in the decrease of cell mo-
bility.32 Based on these experimental results, we
propose that TGFb may modulate the retention of
an appropriate amount of fibroblasts in the wound
area to improve the wound healing process.

ACKNOWLEDGMENTS
AND FUNDING SOURCES
This work is supported by a research grant from
the National Science Council, NSC 101-2311-B-
010-004.

AUTHOR DISCLOSURE AND GHOSTWRITING

Figure 7. Hypothetical wound healing process revealed by proteomics and No competing financial interests exist. The con-
biochemical analysis of fibroblast cells treated with Shiunko. Several biological tent of this article was expressly written by the
functions are involved in the wound healing processes including antioxidant author(s) listed. No ghostwriters were used to write
activity, cell proliferation/viability, collagen secretion, and cell mobility. The
this article.
proteomes detected from the fibroblast cells induced by Shiunko are grouped
in places where the distinct physiological functions are located. The upward
and downward arrows represent upregulation and downregulation of the
distinct physiological functions of the cells. The figure illustrating the wound ABOUT THE AUTHORS
healing process is a combination effect of various biological functions of cells Kin-Fu Chak is the Director of the Institute of
induced by the treatment of Shiunko. To see this illustration in color, the reader
Biomedical Sciences at Mackay Medical College
is referred to the web version of this article at www.liebertpub.com/wound
and a Professor of Biochemistry at National Yang
 TRADITIONAL CHINESE MEDICINE IN WOUND CARE 455

Ming University, Taipei, Taiwan. Chia-Yen Hsiao Ting-Yu Chen contributed on the study of
contributed on the study of Angelica sinensis and Shiunko. Miss Chen obtained her MSc degree in
Lithospermi Radix. Dr. Hsiao obtained his PhD June 2012 at the Institute of Biochemistry and
degree in June 2012 at the Institute of Biochem- Molecular Biology, National Yang Ming Uni-
istry and Molecular Biology, National Yang Ming versity. She is now continuing her PhD degree at
University. Currently he is in military service. the same institute of the university.

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