Korean J Physiol Pharmacol 2019;23(2):103-111

https://doi.org/10.4196/kjpp.2019.23.2.103

Original Article

Beneficial effects of andrographolide in a rat model of

autoimmune myocarditis and its effects on PI3K/Akt pathway
Qi Zhang, Li-qun Hu, Hong-qi Li, Jun Wu, Na-na-Bian, and Guang Yan*
Department of Geriatrics, Anhui Provincial Hospital, The First Affiliated Hospital of University of Science and Technology of China, Anhui Institute of Cardio-
vascular Disease, Hefei 230001, China

ARTICLE INFO
ABSTRACT The study is to investigate effects of andrographolide on experimental
Received April 16, 2018
Revised June 29, 2018
autoimmune myocarditis (EAM). Lewis rats were immunized on day 0 with por-
Accepted July 20, 2018 cine cardiac myosin to establish EAM. The EAM rats were treated with either an-
drographolide (25, 50, 100 mg/kg/day) or vehicle for 21 days. An antigen-specific
*Correspondence splenocytes proliferation assay was performed by using the cells from control rats
Guang Yan immunized with cardiac myosin. Survival rates, myocardial pathology and myocar-
E-mail: [email protected] dial functional parameters (left ventricle end-diastolic pressure, ± dP/dt and left ven-
tricular internal dimension) of EAM rats received andrographolide were significantly
Key Words improved. Andrographolide treatment caused an decrease in the infiltration of CD3+
Akt and CD14+ positive cells in myocardial tissue. Moreover, andrographolide treatment
Andrographolide
caused a reduction in the plasma levels of tumor necrosis factor-alpha, interleukin-17
Autoimmune myocarditis
PI3K (IL-17) and myosin-antibody, and an increase in the level of IL-10 in EAM rats. Oral
administration of andrographolide resulted in the decreased expression of p-PI3K,
p-Akt without any change of PI3K and Akt. Further results indicate andrographolide
significantly inhibited myosin-induced proliferation in splenocytes, and this effect
was inhibited by co-treatment of SC79 (Akt activator). Our data indicate androgra-
pholide inhibits development of EAM, and this beneficial effect may be due to pow-
erful anti-inflammatory activity and inhibitory effect on PI3K/Akt pathway.

INTRODUCTION [3]. Inflammatory response directly deteriorates cardiac function

and induces acute heart failure, chest pain and life threatening
Myocarditis is a non-ischemic heart disease with rapidly pro- arrhythmias [4]. Phosphatidylinositol-3-kinase (PI3K)/Protein
gressive heart failure, arrhythmias, sudden cardiac death, and is a kinase B (Akt) signaling pathway is widely involved in the devel-
major cause of cardiomyopathies-induced death in younger [1]. A opment of inflammatory diseases. The pathway has been identi-
writing group convened by the GBD 2010 Study has estimated the fied as a central mediator of physiological cardiac hypertrophy
burden of myocarditis as a percentage of prevalent heart failure [5]. Dysfunctional PI3K pathway has been broadly observed and
varied by age and region from 0.5% to 4.0% [2]. At present, there demonstrated in angiocardiopathy [6]. In a recent published
is no specific therapy for myocarditis. In most cases, rest cure is report, it has confirmed blocking PI3K/Akt pathway effectively
the main strategy, and supplied with symptomatic therapies for inhibited development of myocarditis in animal models [7].
arrhythmia, heart failure and cardiac shock. Andrographolide is a natural diterpenoid extracted from
Myocarditis is an inflammation of myocardium, and can be Andrographis paniculata , which traditionally used as an herbal
induced by autoimmune disease, infection and cardiotoxic agents medicine in China and Southeast Asia for thousands of years.

This is an Open Access article distributed under the terms Author contributions: Q.Z. and L.Q.H. conceived performed statistical
of the Creative Commons Attribution Non-Commercial analyses, and wrote the paper; Q.Z., H.Q.L., J.W. and N.N.B. performed the
License, which permits unrestricted non-commercial use, distribution, and experiments; G.Y. provided necessary resources, designed the study proto-
reproduction in any medium, provided the original work is properly cited. col and analyzed the data.
Copyright © Korean J Physiol Pharmacol, pISSN 1226-4512, eISSN 2093-3827

www.kjpp.net 103 Korean J Physiol Pharmacol 2019;23(2):103-111

104 Zhang Q et al

It has been previously reported andrographolide displays anti- Assessment of myocardial functions
oxidant, anti-inflammatory, anti-cancer, hypoglycemic, and anti-
virus activity [8]. Moreover, andrographolide has been concluded To evaluate myocardial function, all rats were anesthetized
to attenuate pathological cardiac hypertrophy in high fat diet fed with 2% halothane in oxygen during the surgical procedure. A
mice, and exhibits anti-inflammatory and anticoagulant proper- catheter-tip transducer (Miller SPR-524; Miller Instruments,
ties by inhibiting Mitogen-activated protein kinase (MAPK) and Houston, TX) was introduced into the left ventricle through the
nuclear transcription factor-B (NF-B) pathways in LPS-treated right carotid artery. Cardiac function parameters, including left
cardiac microvascular endothelial cells [9,10]. It is of interest to ventricle end-diastolic pressure (LVEDP), heart rate (HR), and ±
see andrographolide inhibits PI3K/Akt pathway in animal models dP/dt were measured and recorded. An echocardiography system
and cultured cells, inducing a lot of pharmacological actions, such (NEMIO SSA-550A; Toshiba Medical Systems) with a dynami-
as inhibiting tumor cells growth, migration, invasion, and modu- cally focused 14 MHz linear-array transducer and 7 MHz sector
lating the innate and adaptive immune responses [11,12]. How- transducer was used. Measurements were performed for at least
ever, it is still unclear whether andrographolide has a protective ten beats in a blinded fashion, and repeated twice. Left ventricular
effect on myocarditis. Experimental autoimmune myocarditis internal dimension in diastole (LVDD) and left ventricular inter-
(EAM) in rats is a well known model characterized by extensive nal dimension in systole (LVDS) were measured. Ejection fraction
myocardial necrosis, heart failure, which is extremely similar to (EF) was calculated according to the formula: EF (%) = (LVDd3–
that observed in clinical patients. Moreover, EAM shows progress LVDs3)/LVDd3. Following that, the rats were sacrificed and hearts
into clinic pathological status (enlargement of heart, dilatation were removed, weighed and harvested.
of ventricles, diffuse and extensive myocardial inflammation),
which is similar to dilated cardiomyopathy in chronic phase [13]. Anti-cardiac myosin antibody and inflammatory
Therefore, in present study, we used EAM model to evaluate the cytokines levels
efficacy of andrographolide.
Blood was collected from femoral artery of rats and serum was
prepared. The cytokines levels were measured by using ELISA
METHODS kits according to the instructions. The kits of tumor necrosis
factor-alpha (TNF-alpha), Interleukin-17 (IL-17) and IL-10 were
Animals products of R&D Systems (Minneapolis, MN, USA). Moreover,
the level of anti-cardiac myosin was determined by a previously
The Lewis rats (male, eight weeks old) were purchased from published method [14].
Model Animal Research Center of Nanjing University (Nanjing,
China), and placed in the animal house under controlled condi- Cell proliferation assay
tions of 25 ± 2°C, relative humidity of 60 ± 5% and a light–dark
cycle of 12:12 h. All rats were fed with food pellets and water ad Antigen-specific splenocytes proliferation was examined. In
libitum. The current study was approved by the Animal Ethics brief, splenocytes were isolated from the rats with myocarditis.
Committee of Anhui Provincial Hospital. The cells were cultured in a 96-well plates in RPMI-1640 (Invitro-
gen) supplemented with 10% fetal bovine serum and 1% penicil-
Induction of experimental autoimmune myocarditis lin/streptomycin solution. Following that, the cells were pretreat-
ed with andrographolide (10, 20 and 40 M) or Los (10 M) for
Purified porcine cardiac myosin (Sigma Chemical Co., St. 60 min, and then they were treated with porcine heart myosin (20
Louis, MO, USA) was dissolved in 0.01 M phosphate-buffered g/ml) for 48 h. The proliferation of splenocytes was determined
saline (PBS) and emulsified with an equal volume of complete by a MTT assay.
Freund’s adjuvant (CFA, Difco, Sparks, MD, USA) supplemented
with mycobacterium tuberculosis H37RA (10 mg/ml, Difco). Histological analysis
0.2 ml of emulsion was injected into the footpads of rats subcu-
taneously. For the control group, the rats received CFA in same The excised myocardium was kept in formalin and then the
manner. Following injection, a total of 56 rats were divided into sections were embedded in paraffin. The heart tissue was sec-
several groups: control group (n = 6), EAM group (n = 10), an- tioned (4 m thick) and stained with hematoxylin and eosin. The
drographolide (purity>98%; Sigma-Aldrich, St. Louis, MO, USA) sections were scored for myocarditis as follows: 0, no infiltration
groups (25, 50, 100 mg/kg, n = 3×10) and losartan (Los, Aladdin or necrosis;1, between 1% and 25% infiltration or necrosis per sec-
biotechnology co., LTD; ShangHai, China) group (10 mg/kg, n = tion; 2, between 26% and 50% infiltration or necrosis per section;
10). After immunization, the rats were orally administrated with 3, between 51% and 74% infiltration or necrosis per section; and
drugs or vehicle (control group) for 21 days. 4, between 75% and 100% infiltration or necrosis per section. The

Korean J Physiol Pharmacol 2019;23(2):103-111 https://doi.org/10.4196/kjpp.2019.23.2.103

Cardioprotective effects of andrographolide 105

sections were examined by light microscopy and all histological Biotechnology). After incubation with HRP-coupled secondary
evaluations were done by our laboratory technician who didn’t antibody (Santa Cruz Biotechnology), the bands were visualized
know the groups information. Moreover, the area of inflamma- with a chemiluminescence developing agents (Amersham Biosci-
tory cells was evaluated by image pro plus software (6.0 version), ences, UK). GAPDH was probed in the blots as internal control
which were showed as the ratio of area of inflammatory cells to for loading. Results were semi-quantified as optical density band
that of total area. area by the Image J.

Immunohistochemical analysis Statistical analysis

Formalin-fixed and paraffin-embedded cardiac tissue sections Data are presented as mean ± S.D. Statistical analysis of differ-
were used for immunostaining. Briefly, the sections were incubat- ences between the groups was performed using one-way analysis
ed with anti-CD3+ (BD Biosciences, San Jose, CA, USA), or anti- of variance, followed by Tukey’s multiple comparison test (SPSS
CD14+ (BD Biosciences) antibodies, followed by incubation with 15.0, SPSS Inc., Chicago, IL, USA). Kruskal–Wallis test was used
HRP-linked rabbit anti-mouse IgG. The reaction products were to analyze survival times, as multiple groups were used. A value
visualized by using 0.5% diaminobenzidine and 0.03% hydrogen of p < 0.05 was considered statistically.
peroxide. The numbers of positive-stained cells were determined
in ten randomly selected fields by a professional staff, who didn’t
know the grouped details. RESULTS
Western blotting Effects of andrographolide on mortality, myocardial
functions of EAM rats
Myocardial tissue samples were homogenized with lysis buf-
fer, and the protein was extracted. Protein concentrations were As seen in Fig. 1, survival curves showed protective effects of
measured by the bicinchoninic acid method. The samples were andrographolide on EAM rats. Myocarditis rats given vehicle
separated by SDS-PAGE and identified with following antibodies: had five deaths after myosin injection, whereas those treated with
PI3K, phosphorylated PI3K (p-PI3K), Akt and p-Akt (Santa Cruz andrographolide (50, 100 mg/kg) showed three and one deaths,

Fig. 1. Effects of andrographolide on mortality of EAM rats. (A) Survival curves of EAM rats. (B) Representative echocardiogram of M-mode on day
21. Control group includes six rats, whereas other groups include ten rats for each. Con: control; EAM: experimental autoimmune myocarditis; Los:
losartan.

www.kjpp.net Korean J Physiol Pharmacol 2019;23(2):103-111

106 Zhang Q et al

respectively. The 21-day mortality was 50.0% in the EAM group, Effects of andrographolide on myocardial
and 10.0% in andrographolide (100 mg/kg) group. Similar obser- histopathology of EAM rats
vation was presented in Los group compared with that of EAM
group. As shown in Table 1, the cardiac function parameters, such As seen in Fig. 2, there were increased infiltration of inflam-
as LVEDP, LVDS, HW/BW were significantly higher, and ±dP/ matory cells in EAM groups compared with control group (Fig.
dt & EF were significantly lower in EAM model compared with 2, p < 0.01). However, treatment of andrographolide (50, 100 mg/
control (p < 0.05). However, compared with vehicle-treated EAM kg) and Los obviously inhibited infiltration of inflammatory cells
rats, treatment of andrographolide or Los significantly decreased in myocardial tissue (p < 0.01). Our data suggest andrographolide
LVEDP, LVDS, HW/BW, and increased ± dP/dt and EF (p < 0.05). effectively improves cardiac pathological performance of EAM

Table 1. Parameters of cardiac function at day 21 after myosin injection

Andrographolide (mg/kg) Los (mg/kg)

Parameters Con EAM
25 50 100 10
##
LVEDP mm Hg 3.29 ± 1.2 22.19 ± 6.2 19.24 ± 5.2 14.53 ± 4.9** 11.33 ± 5.2** 9.29 ± 6.6**
+dP/dt mm Hg/s 8264 ± 1223 2128 ± 366## 2917 ± 423* 3735 ± 786** 5432 ± 878** 6425 ± 1005**
–dP/dt mm Hg/s 7864 ± 1223 2001 ± 298## 2719 ± 379* 3354 ± 425** 4725 ± 564** 5508 ± 728**
HR beats/min 327 ± 15.2 315 ± 18.7 309 ± 22.4 312 ± 17.7 325 ± 17.4 321 ± 14.5
LVDD mm 7.9 ± 0.8 7.5 ± 1.2 7.4 ± 1.0 7.2 ± 1.1 7.6 ± 1.5 7.5 ± 0.9
LVDS mm 4.2 ± 0.6 6.9 ± 0.9## 6.2 ± 0.7 5.4 ± 1.3 4.7 ± 0.6** 4.6 ± 0.8**
EF % 82.0 ± 4.6 20.2 ± 2.6## 43.6 ± 7.5** 64.4 ± 5.7** 72.9 ± 3.7** 78.4 ± 3.3**
HW/BW ‰ 3.5 ± 0.4 6.1 ± 0.7## 5.5 ± 0.5* 4.8 ± 1.4** 4.1 ± 0.8** 4.4 ± 1.3**

Results are presented as the mean ± S.D. N = 6-9. LVEDP, left ventricular end-diastolic pressure; ±dP/dt, rate of intra-ventricular pressure
rise and decline; HR, heart rate; LVDD, left ventricular dimension in diastole; LVDS, left ventricular dimension in systole; EF, ejection
fraction; Con: control; EAM: experimental autoimmune myocarditis; Los: losartan. ##p < 0.01, compared with Con; *p < 0.05, **p < 0.01,
compared with EAM.

Fig. 2. Effects of andrographolide on myocardial histopathology of EAM rats. (A) Con group; (B) EAM group; (C) andrographolide 25 mg/kg;
(D) andrographolide 50 mg/kg; (E) andrographolide 100 mg/kg; (F) losartan 10 mg/kg; (G) pathological scores were evaluated by professional staff.
Results are presented as the mean ± S.D. N = 6-9. Con: control; EAM: experimental autoimmune myocarditis; Los: losartan. ##p < 0.01, compared with
Con; *p < 0.05, **p < 0.01, compared with EAM.

Korean J Physiol Pharmacol 2019;23(2):103-111 https://doi.org/10.4196/kjpp.2019.23.2.103

Cardioprotective effects of andrographolide 107

rats. tigated effects of andrographolide on infiltration of T cells and

monocytes using immunohistochemical analysis. As shown in
Effects of andrographolide on myocardial Fig. 3 and Fig. 4, compared with rats in control group, vehicle-
inflammatory response in EAM rats treated EAM rats showed increased numbers of positive CD3+
and CD14+ cells in the tissue (p < 0.01), suggesting enhanced
Histological examination showed a lot of inflammatory cells myocardial inflammatory response. However, andrographolide
infiltrated in the myocardial tissue. Therefore, we further inves- significantly reduced the numbers of positive CD3+ and CD14+

Fig. 3. Effects of andrographolide on the numbers of CD3+ positive cells in myocardium of EAM rats. (A) Con group; (B) EAM group; (C) an-
drographolide 25 mg/kg; (D) andrographolide 50 mg/kg; (E) andrographolide 100 mg/kg; (F) the numbers of CD3+ positive cells were determined in
ten randomly selected fields by a professional staff, who didn’t know grouped details. Results are presented as the mean ± S.D. N = 6-9. Con: control;
EAM: experimental autoimmune myocarditis; ##p < 0.01, compared with Con; *p < 0.05, **p < 0.01, compared with EAM.

Fig. 4. Effects of andrographolide on the numbers of CD14+ positive cells in myocardium of EAM rats. (A) Con group; (B) EAM group; (C) an-
drographolide 25 mg/kg; (D) andrographolide 50 mg/kg; (E) andrographolide 100 mg/kg; (F) the numbers of CD14+ positive cells were determined in
ten randomly selected fields by a professional staff, who didn’t know grouped details. Results are presented as the mean ± S.D. N = 6-9. Con: control;
EAM: experimental autoimmune myocarditis; ##p < 0.01, compared with Con; *p < 0.05,**p < 0.01, compared with EAM.

www.kjpp.net Korean J Physiol Pharmacol 2019;23(2):103-111

108 Zhang Q et al

cells in myocardial tissue (p < 0.01). Moreover, we have detected MTT assay to examine effects of andrographolide on myosin-
plasma inflammatory cytokines and anti-myosin antibody in all induced proliferation in splenocytes. As seen in Fig. 7, myosin
rats by ELISA kits. As seen in Fig. 5, compared with EAM group, significantly stimulated splenocytes proliferation (p < 0.01), and
andrographolide oral administration significantly reduced the the effect of myosin was significantly inhibited by co-treatment
levels of TNF-alpha, IL-17 and anti-myosin (p < 0.01). In contrast, of andrographolide (p < 0.01). However, an Akt activator, SC79,
the production of IL-10 was significantly elevated in the an- significantly inhibited andrographolide-induced inhibitory ef-
drographolide-treated group compared with the vehicle-treated fects on proliferation of splenocytes (Fig. 7, p < 0.05). Finally, the
group (p < 0.01). data from western blotting indicated SC79 treatment effectively
increased the expression of p-Akt in andrographolide-treated
Effects of andrographolide on PI3K/Akt signaling cells (Fig. 7, p < 0.01).
pathway in EAM rats

As seen in Fig. 6, expressions of p-PI3K and p-Akt were obvi- DISCUSSION

ously increased in vehicle-treated EAM rats compared with that
of control rats (p < 0.01). In the andrographolide groups, the In this study, we investigated the effects of andrographolide in
myocardial levels of p-PI3K and p-Akt were significantly reduced a rat model of EAM. Treatment with andrographolide reduced
compared with that of EAM group (p < 0.01). Moreover, we found mortality rates, improved cardiomyopathology and cardiac func-
andrographolide treatment had no effects on the total expression tions. Our result is consistent with previous reports that conclud-
of PI3K and Akt in EAM rats. ed andrographolide showed a well protective action in cardio-
vascular diseases. In previous studies, Zhang et al. [15] reported
Effects of andrographolide on myosin-induced andrographolide attenuates LPS-induced cardiac malfunctions.
splenocytes proliferation Hsieh et al. [10] found andrographolide displays inhibitory ef-
fects on cardiac hypertrophy and myocardial apoptosis in high-
To study whether andrographolide directly inhibits cardiac fat diet fed mice. Woo et al. [16] have confirmed andrographolide
myosin-induced activation in lymphocytes, we performed a protected cardiomyocytes against hypoxia/reoxygenation injury.

Fig. 5. Effects of andrographolide on the levels of inflammatory cytokines and myosin antibody in EAM rats. (A) TNF-alpha; (B) IL-10; (C) IL-17; (D)
myosin antibody. Results are presented as the mean ± S.D. N = 6-9. EAM: experimental autoimmune myocarditis; ##p < 0.01, compared with Control;
*p < 0.05, **p < 0.01, compared with EAM.

Korean J Physiol Pharmacol 2019;23(2):103-111 https://doi.org/10.4196/kjpp.2019.23.2.103

Cardioprotective effects of andrographolide 109

Fig. 6. Effects of andrographolide on PI3K/Akt pathway. (A) Representative stripes; (B, C) the stripes were quantitatively analyzed by Image J. Re-
sults are showed as the mean ± S.D. N = 6-9. Con: control; EAM: experimental autoimmune myocarditis. ##p < 0.01, compared with Con; **p < 0.01,
compared with EMA.

Fig. 7. Effects of andrographolide on proliferation of myosin-treated splenetic cells. (A) Splenetic cells were collected from EAM rats, and treated
with andrographolide in the presence and absence of SC79 for 48 h. Myosin-specific proliferative responses were measured by MTT colorimetric tech-
nique. (B1) typical p-Akt stripes; (B2) the stripes were quantitatively analyzed by image j. Results are presented as the mean ± S.D. N = 3. Con: control;
EAM: experimental autoimmune myocarditis. ##p < 0.01, compared with splenetic cells without treatment; *p < 0.05, **p < 0.01, compared with sple-
netic cells only treated with myosin (20 g/ml); △p < 0.05, compared with the cells treated with myosin and andrographolide (40 M).

Our data firstly prove protective action of andrographolide in the resided immune cells or the circulating cells that migrated to
EAM model. myocardium [3]. Some studies showed cardiac antigen-specific
Inflammatory response play important roles in the progression naïve T cells, especially those specific of -myosin heavy chain
of cardiovascular diseases. At the beginning of EAM, the injected peptides, become activated and differentiate into expanded clones
myosin attacks myocardial tissue, and thereby activates cardiac of effector T cells under various conditions, such as cardiac in-

www.kjpp.net Korean J Physiol Pharmacol 2019;23(2):103-111

110 Zhang Q et al

fection and/or genetic variations in peripheral tolerance [17]. It Andrographolide has been proposed to be an anti-inflammatory
has been previously showed monocyte control T cell responses medicine with multiple targets, including NF-B, MAPKs and
against cardiac myosin in myocarditis animal models. However, JAK/STAT pathways [8]. Moreover, andrographolide has been
exaggerated monocyte activation by cardiac myosin peptide TLR reported to inhibit inflammatory response in TNF-alpha-treated
ligands in pathogenesis of myocarditis represents a two-edged HUVEC cells by down-regulation of PI3K/Akt pathway, and at-
sword. Monocyte-derived cytokines not only promote develop- tenuate TNF-alpha-induced ICAM-1 expression with an PI3K/
ment of Th17 cells, but also result in the activation of TGF-- Akt pathway-dependent manner [34,35]. To confirm the role of
mediated cardiac fibrosis [18]. Moreover, inflammatory cytokines PI3K/Akt pathway in protective bioactivity of andrographolide
secreted by monocytes and T cells, directly damage the functions against EAM, we analyzed expression of PI3K, p-PI3K, Akt and
of cardiomyocytes. IL-17 has been confirmed to promote cardiac p-Akt in the cardiac tissue. The results showed andrographolide
fibrosis and inhibit myocardial diastolic function [19-21]. IL-10 significantly reduced cardiac levels of p-PI3K and p-Akt without
knockout increase the sensitivity of left ventricular dysfunction to any change of PI3K and Akt. Interestingly, we noted SC79 sig-
pressure overload, while IL-10 treatment improve left ventricular nificantly reduced andrographolide-induced inhibitory effects on
functions in rats with myocardial infarction. These effects can be proliferation of the splenocytes.
explained as follows: IL-17 regulates cardiomyocytes apoptosis, In conclusion, our results demonstrate protective effects of
while IL-10 suppresses production of inflammatory cytokines andrographolide on EAM model were primarily associated with
in rat cardiomyocytes, and attenuates TNF-alpha-induced car- suppression of cardiac inflammation and blockade of PI3K/Akt
diomyocytes apoptosis [22,23]. Moreover, TNF-alpha induces pathway. Our findings indicate the potential value of androgra-
apoptosis in cardiomyocytes and promotes cardiac inflam- pholide for treating human myocarditis.
mation and fibrosis [24,25]. In present study, andrographolide
treatment significantly inhibited infiltration of CD3+ positive T
cells and CD14+ positive monocytes in EAM rats. Moreover, an- ACKNOWLEDGEMENTS
drographolide reduced circulating levels of TNF-alpha, IL-17 and
myosin-antibody. Our data suggest andrographolide improves This work was supported by the Anhui Province Nature Sci-
myosin-induced inflammation in EAM rats. It should be noted ence Foundation in the University (kj2013z125).
that andrographolide has an excellent anti-inflammatory activ-
ity. In vitro study, andrographolide has been reported to suppress
production of a lot of inflammatory cytokines, chemokines and CONFLICTS OF INTEREST
enzymes in immune cells (macrophages, fibroblasts) and human
bronchial epithelial cells [8]. In vivo studies, andrographolide has The authors declare no conflicts of interest.
been confirmed to exhibit powerful anti-inflammatory effects in
animal models of pulmonary fibrosis, allergic airway and diabetic
nephropathy [26-28]. REFERENCES
PI3K/Akt is an important signaling pathway in the regulation
of inflammatory response. The pathway comprises of two main 1. Drory Y, Turetz Y, Hiss Y, Lev B, Fisman EZ, Pines A, Kramer MR.
driving molecules: PI3K and Akt. PI3K could be activated by re- Sudden unexpected death in persons less than 40 years of age. Am J
ceptor tyrosine kinases and G protein-coupled receptors, forming Cardiol. 1991;68:1388-1392.
p-PI3K. Following PI3K activation and serial cascade reactions, 2. Cooper LT Jr, Keren A, Sliwa K, Matsumori A, Mensah GA. The
global burden of myocarditis: part 1: a systematic literature review
Akt is phosphorylated, and eventually triggers multiple down-
for the global burden of diseases, injuries, and risk factors 2010
stream pathways involved in protein synthesis, cell proliferation, study. Glob Heart. 2014;9:121-129.
metabolism and survival [5]. Previous studies confirmed cardiac- 3. Sagar S, Liu PP, Cooper LT Jr. Myocarditis. Lancet. 2012;379:738-
restricted over-expression of either PI3K or its upstream IGF- 747.
1 receptor resulted in increased cardiomyocytes size and larger 4. Elamm C, Fairweather D, Cooper LT. Pathogenesis and diagnosis of
hearts. Constitutive activation of Akt increased cardiomyocytes myocarditis. Heart. 2012;11:835-840.
size and led to concentric left ventricle hypertrophy, while 5. Ghigo A, Li M. Phosphoinositide 3-kinase: friend and foe in cardio-
Akt knockout mice displayed smaller heart [29-32]. Moreover, vascular disease. Front Pharmacol. 2015;6:169.
cardiac-specific activation of Akt promoted vascular endothelial 6. Yu P, Zhang Y, Li C, Li Y, Jiang S, Zhang X, Ding Z, Tu F, Wu J, Gao
X, Li L. Class III PI3K- mediated prolonged activation of autophagy
growth factor and angiopoietin-2 expression in cardiomyocytes,
plays a critical role in the transition of cardiac hypertrophy to heart
and ultimately increased myocardial capillary density and physi-
failure. J Cell Mol Med. 2015;19:1710-1719.
ological hypertrophic remodeling [33]. PI3K inhibitor ameliorates 7. Liu HS, Zhang J, Guo JL, Lin CY, Wang ZW. Phosphoinositide 3-ki-
the symptom of myosin-induced myocarditis in mice [7], suggest- nase inhibitor LY294002 ameliorates the severity of myosin-induced
ing the pathway is the potential therapeutic target of myocarditis. myocarditis in mice. Curr Res Transl Med. 2016;64:21-27.

Korean J Physiol Pharmacol 2019;23(2):103-111 https://doi.org/10.4196/kjpp.2019.23.2.103

Cardioprotective effects of andrographolide 111

8. Lim JC, Chan TK, Ng DS, Sagineedu SR, Stanslas J, Wong WS. An- Interleukin-17A mediates cardiomyocyte apoptosis through Stat3-
drographolide and its analogues: versatile bioactive molecules for iNOS pathway. Biochim Biophys Acta. 2016;1863:2784-2794.
combating inflammation and cancer. Clin Exp Pharmacol Physiol. 23. Damås JK, Aukrust P, Ueland T, Odegaard A, Eiken HG, Gullestad L,
2012;39:300-310. Sejersted OM, Christensen G. Monocyte chemoattractant protein-1
9. He CL, Yi PF, Fan QJ, Shen HQ, Jiang XL, Qin QQ, Song Z, Zhang C, enhances and interleukin-10 suppresses the production of inflam-
Wu SC, Wei XB, Li YL, Fu BD. Xiang-Qi-Tang and its active com- matory cytokines in adult rat cardiomyocytes. Basic Res Cardiol.
ponents exhibit anti-inflammatory and anticoagulant properties by 2001;96:345-352.
inhibiting MAPK and NF-B signaling pathways in LPS-treated rat 24. Chen ZW, Qian JY, Ma JY, Chang SF, Yun H, Jin H, Sun AJ, Zou
cardiac microvascular endothelial cells. Immunopharmacol Immu- YZ, Ge JB. TNF--induced cardiomyocyte apoptosis contributes to
notoxicol. 2013;35:215-224. cardiac dysfunction after coronary microembolization in mini-pigs.
10. Hsieh YL, Shibu MA, Lii CK, Viswanadha VP, Lin YL, Lai CH, J Cell Mol Med. 2014;18:1953-1963.
Chen YF, Lin KH, Kuo WW, Huang CY. Andrographis paniculata 25. Duerrschmid C, Trial J, Wang Y, Entman ML, Haudek SB. Tumor
extract attenuates pathological cardiac hypertrophy and apoptosis necrosis factor: a mechanistic link between angiotensin-II-induced
in high-fat diet fed mice. J Ethnopharmacol. 2016;192:170-177. cardiac inflammation and fibrosis. Circ Heart Fail. 2015;8:352-361.
11. Lee YC, Lin HH, Hsu CH, Wang CJ, Chiang TA, Chen JH. Inhibito- 26. Guan SP, Kong LR, Cheng C, Lim JC, Wong WS. Protective role of
ry effects of andrographolide on migration and invasion in human 14-deoxy-11,12-didehydroandrographolide, a noncytotoxic ana-
non-small cell lung cancer A549 cells via down-regulation of PI3K/ logue of andrographolide, in allergic airway inflammation. J Nat
Akt signaling pathway. Eur J Pharmacol. 2010;632:23-32. Prod. 2011;74:1484-1490.
12. Wang W, Wang J, Dong SF, Liu CH, Italiani P, Sun SH, Xu J, Bora- 27. Ji X, Li C, Ou Y, Li N, Yuan K, Yang G, Chen X, Yang Z, Liu B,
schi D, Ma SP, Qu D. Immunomodulatory activity of androgra- Cheung WW, Wang L, Huang R, Lan T. Andrographolide amelio-
pholide on macrophage activation and specific antibody response. rates diabetic nephropathy by attenuating hyperglycemia- mediated
Acta Pharmacol Sin. 2010;31:191-201. renal oxidative stress and inflammation via Akt/NF-B pathway.
13. Veeraveedu PT, Watanabe K, Ma M, Thandavarayan RA, Palani- Mol Cell Endocrinol. 2016;437:268-279.
yandi SS, Yamaguchi K, Suzuki K, Kodama M, Aizawa Y. Compara- 28. Yin JN, Li YN, Gao Y, Li SB, Li JD. Andrographolide plays an im-
tive effects of torasemide and furosemide in rats with heart failure. portant role in bleomycin-induced pulmonary fibrosis treatment.
Biochem Pharmacol. 2008;75:649-659. Int J Clin Exp Med. 2015;8:12374-12381.
14. Liu X, Zhang X, Ye L, Yuan H. Protective mechanisms of berber- 29. Condorelli G, Drusco A, Stassi G, Bellacosa A, Roncarati R, Iaccari-
ine against experimental autoimmune myocarditis in a rat model. no G, Russo MA, Gu Y, Dalton N, Chung C, Latronico MV, Napoli C,
Biomed Pharmacother. 2016;79:222-230. Sadoshima J, Croce CM, Ross J Jr. Akt induces enhanced myocar-
15. Zhang J, Zhu D, Wang Y, Ju Y. Andrographolide attenuates LPS- dial contractility and cell size in vivo in transgenic mice. Proc Natl
induced cardiac malfunctions through inhibition of IB phosphor- Acad Sci U S A. 2002;99:12333-12338.
ylation and apoptosis in mice. Cell Physiol Biochem. 2015;37:1619- 30. DeBosch B, Treskov I, Lupu TS, Weinheimer C, Kovacs A, Courtois
1628. M, Muslin AJ. Akt1 is required for physiological cardiac growth.
16. Woo AY, Waye MM, Tsui SK, Yeung ST, Cheng CH. Androgra- Circulation . 2006;113:2097-2104.
pholide up-regulates cellular-reduced glutathione level and protects 31. McMullen JR, Shioi T, Huang WY, Zhang L, Tarnavski O, Bisping
cardiomyocytes against hypoxia/reoxygenation injury. J Pharmacol E, Schinke M, Kong S, Sherwood MC, Brown J, Riggi L, Kang PM,
Exp Ther. 2008;325:226-235. Izumo S. The insulin-like growth factor 1 receptor induces physi-
17. Lichtman AH. The heart of the matter: protection of the myocar- ological heart growth via the phosphoinositide 3-kinase(p110α)
dium from T cells. J Autoimmun. 2013;45:90-96. pathway. J Biol Chem. 2004;279:4782-4793.
18. Myers JM, Cooper LT, Kem DC, Stavrakis S, Kosanke SD, Shevach 32. Shioi T, Kang PM, Douglas PS, Hampe J, Yballe CM, Lawitts J,
EM, Fairweather D, Stoner JA, Cox CJ, Cunningham MW. Cardiac Cantley LC, Izumo S.The conserved phosphoinositide 3-kinase
myosin-Th17 responses promote heart failure in human myocardi- pathway determines heart size in mice. EMBO J. 2000;19:2537-2548.
tis. JCI Insight. 2016;1(9):e85851. 33. Shiojima I , Sato K, Izumiya Y, Schiekofer S, Ito M, Liao R, Colucci
19. Liu Y, Zhu H, Su Z, Sun C, Yin J, Yuan H, Sandoghchian S, Jiao Z, WS, Walsh K. Disruption of coordinated cardiac hypertrophy and
Wang S, Xu H. IL-17 contributes to cardiac fibrosis following ex- angiogenesis contributes to the transition to heart failure. J Clin
perimental autoimmune myocarditis by a PKC/Erk1/2/NF-B- Invest. 2005;115:2108-2118.
dependent signaling pathway. Int Immunol . 2012;24:605-612. 34. Chen HW, Lin AH, Chu HC, Li CC, Tsai CW, Chao CY, Wang CJ,
20. Stumpf C, Seybold K, Petzi S, Wasmeier G, Raaz D, Yilmaz A, Anger Lii CK, Liu KL. Inhibition of TNF--Induced inflammation by
T, Daniel WG, Garlichs CD. Interleukin-10 improves left ventricular andrographolide via down-regulation of the PI3K/Akt signaling
function in rats with heart failure subsequent to myocardial infarc- pathway. J Nat Prod. 2011;74:2408-2413.
tion. Eur J Heart Fail. 2008;10:733-739. 35. Lu CY, Yang YC, Li CC, Liu KL, Lii CK, Chen HW. Androgra-
21. Yu Y, Zhang ZH, Wei SG, Chu Y, Weiss RM, Heistad DD, Felder pholide inhibits TNF-induced ICAM-1 expression via suppression
RB. Central gene transfer of interleukin-10 reduces hypothalamic of NADPH oxidase activation and induction of HO-1 and GCLM
inflammation and evidence of heart failure in rats after myocardial expression through the PI3K/Akt/Nrf2 and PI3K/Akt/AP-1 path-
infarction. Circ Res. 2007;101:304-312. ways in human endothelial cells. Biochem Pharmacol. 2014;91:40-
22. Su SA, Yang D, Zhu W, Cai Z, Zhang N, Zhao L, Wang JA, Xiang M. 50.

www.kjpp.net Korean J Physiol Pharmacol 2019;23(2):103-111