Anti-inflammatory Activities of Pineapple (Ananas comosus) Core

Extract in Lipopolysaccharide-induced RAW264.7 Cell Line

Hanna Sari Widya Kusuma

1,* a

, Hartini Tiono

, Philips Onggowidjaja

Selonan Susang Obeng

, Wahyu Widowati

2* e

, Cintani Dewi Wahyuni

Cahyaning Riski Wijayanti

, Muhamad Aldi Maulana

, Tri Handayani

and Rizal Rizal

1,3 j

Biomolecular and Biomedical Research Center, Aretha Medika Utama, Jl. Babakan Jeruk II No. 9, Bandung,

West Java, Indonesia

Faculty of Medicine, Maranatha Christian University, Jl. Prof. Drg. Surya Sumantri No. 65, Bandung,

West Java, Indonesia

Biomedical Engineering, Department of Electrical Engineering, Faculty of Engineering, Universitas Indonesia,

Jl. Kampus UI, Depok, West Java, Indonesia

wahyu_w60@yahoo.com, cintanidewi@gmail.com, cahyaningwidodo@gmail.com, aldimaulana.srl@gmail.com,

mbaktrihandayani@gmail.com, rizal_biotek@yahoo.com

Keywords: Anti-inflammatory, IL-1β, Pineapple, RAW 264.7 Cell Lines.

Abstract: Inflammation is a biological response process by the immune system that may induce acute/chronic

inflammatory and leading tissue damage or diseases. Pineapple (Ananas comosus) cores that have been

investigated for anti-inflammatory properties and immunomodulator. This research aims to evaluate the anti-

inflammatory potency of pineapple core extract (PCE) in lipopolysaccharide-induced macrophage cells

(RAW 264.7). The viability assay of PCE was determined by MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-

carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) to ensure the safe and non-toxic concentration in

RAW 264.7 cells. The pro-inflammatory induction of cells using 200 μL of lipopolysaccharide (LPS). Levels

of PGE-2 and pro-inflammatory cytokines IL-1β and TNF-α levels were measured using enzyme-linked

immunosorbent assay. RESULTS: PCE 4 and 20 µg/mL showed high viability (>90%) with the values

95.03% and 92.94%, respectively. PCE 20 µg/mL showed the lower of PGE-2 and TNF-α levels (507.68

pg/mL; 345.90 pg/mL) compared to PCE 4 µg/mL (795.37 pg/mL; 474.19 pg/mL) and positive control (870.48

pg/mL; 581.71 pg/mL). In IL-1β level, PCE 20 µg/mL showed the lower (217.63 pg/mL) compared to PCE 4

pg/mL (350.78 pg/mL) and positive control (433.53 pg/mL). Pineapple core extract has beneficial for anti-

inflammatory by downregulating inflammatory mediators including PGE-2, TNF-α, and IL-1β in LPS-

induced RAW 264.7 cell lines.

INTRODUCTION

Inflammation is an immune system reaction that can

https://orcid.org/0000-0002-7422-0036

https://orcid.org/0000-0002-8050-1707

https://orcid.org/0000-0002-7161-9762

https://orcid.org/0000-0003-0608-3516

https://orcid.org/0000-0002-5401-7794

https://orcid.org/0000-0002-7764-0482

https://orcid.org/0000-0002-3397-099X

https://orcid.org/0000-0003-4724-7548

https://orcid.org/0000-0001-9186-9841

https://orcid.org/0000-0003-2783-0672

Corresponding author

be caused by a several factors, including damaged

cells, pathogens, and toxic substances. These factors

can cause acute and/or chronic inflammatory

Kusuma, H., Tiono, H., Onggowidjaja, P., Obeng, S., Widowati, W., Wahyuni, C., Wijayanti, C., Maulana, M., Handayani, T. and Rizal, R.

Anti-inﬂammatory Activities of Pineapple (Ananas comosus) Core Extract in Lipopolysaccharide-induced RAW264.7 Cell Line.

DOI: 10.5220/0010744300003113

In Proceedings of the 1st International Conference on Emerging Issues in Technology, Engineering and Science (ICE-TES 2021), pages 58-64

ISBN: 978-989-758-601-9

responses in the pancreas, liver, heart, lung, brain,

kidney, reproductive system, and intestinal tract,

which could result in tissue damage or diseases (Chen

et al., 2018).

Inflammatory cytokines are produced by activated

macrophages, such as interleukins (ILs), tumor

necrosis factor (TNF-α), inflammatory mediators like

nitric oxide (NO), and prostaglandins (PGs), which

play a protective role for the host in inflammatory

conditions and also preserving normal cellular

conditions (Lee et al., 2017; Saanin et al., 2020). The

activation of macrophages causes the release of a

variety of chemicals, including Nitric Oxide (NO),

reactive oxygen species (ROS), prostaglandin E

(PGE), and Interleukin (IL)-1β, Interleukin (IL)-6,

cyclooxygenase-2 (COX-2), and

tumor necrosis factor

(TNF)-α (Widowati et al., 2016;

Novilla et al., 2017;

Lee et al., 2017; Laksmitawati et al., 2017). Many

chronic diseases have been linked to the inducible

forms of nitric oxide synthase (NOS) and

cyclooxygenase (COX), which are responsible for

raising NO and prostaglandins (PGs) levels,

respectively (Widowati et al., 2021). Thus, the

suppression of pro-inflammatory mediators can be an

effective indicator of anti-inflammatory drugs

(Saanin et al., 2020; Widowati et al., 2021).

Anti-inflammatory medications, both steroidal

and non-steroidal, are used in conventional treatment

for inflammatory diseases. However, the limitations

and risks associated with conventional therapy have

led people to explore alternative measures such as

medicinal plants for the treatment of inflammatory

diseases (Kargutkar and Brijesh, 2017).

Ananas comosus (L.) Merr. which is known as

pineapple is a species of tropical plant that belongs to

the Bromeliaceae family (Rahman et al., 2020). A.

comosus has a various compounds from several parts

of the plant, including alkaloids, anthraquinones,

bromelain, cardiac glycoside, coumarins, flavonoids,

glycoside, inulin, naphthoquinones, phenols,

phytosterols, polyphenols, quinine, saponin, steroids,

sterols, tacorins, terpenoids, tannins, and triterpenes

(Rahman et al., 2020). A. comosus can prevent

undesirable inflammatory processes and also has anti-

inflammatory activity (Yatoo et al., 2018). Bromelain

is a bioactive compound and as a major protease

enzyme found in A. comosus stems that demonstrate

anti-thrombotic, anti-inflammatory, and anti-

edematous (Ramli et al., 2018). Bromelain also has

anti-cancer properties and facilitates cell death by

apoptosis (Pavan et al., 2012). Pineapple core extract

(PCE) exhibit as antioxidant potent by 2,2-diphenyl-

1-picryl-hydrazyl-hydrate (DPPH) scavenging

activity (Vrianty et al., 2019).

However, the previous studies were focused on

the fruit, peels, and leaves of pineapple (A. comosus)

but there were limited studies on the core of the fruit.

This study was purpose to evaluates the anti-

inflammatory activity of PCE through the inhibitory

activity of pro-inflammatory mediators including

PGE-2, IL-1β, and TNF-α on LPS-induced RAW 264

cells.

METHODS AND MATERIALS

2.1 Extract Preparation

A. comosus plants were obtained from Tambaksari

village, Cagak district, Subang, West Java, Indonesia.

Plant identification was done at Herbarium Bandung

Laboratory, School of Biological Sciences, Bandung

Institute of Technology. The preparation of A.

comosus ethanolic extract based on Vrianty et al.

(2019) method. A.

comosus

were sorted, washed,

weighed in wet weight,

dried in a food dehydrator,

weighed in dry weight, and then crushed into powder

form (core crude drug). And then, the core crude drug

was extracted using maceration techniques with a

70% ethanol solvent. Every 24-hour, the filtrate was

until the ethanol filtrate turned colorless. Before being

used for assays, PCE was stored at 20 ºC (Vrianty et

al., 2019).

2.2 Raw264.7 Cells Culture

The RAW 264.7 (ATCC®TIB-71™) murine

macrophage cell line was obtained from the

Biomolecular and Biomedical Research Center,

Aretha Medika Utama. RAW 264.7 cells were grown

in Dulbecco's Modified Eagle Medium (DMEM)

(Biowest, L0104) supplemented with 10% fetal

bovine serum (FBS) (Biowest, S1810) and 1%

antibiotic–antimycotic (Gibco, 15240062). The cells

were incubated at 37 ºC and 5% CO

in the

humidified atmosphere until confluent (80%–90%).

Trypsin-EDTA 0.25% (Gibco, 25200072) was used

to harvest the cells which were then seeded on plates

for the assays (Sandhiutami et al., 2017;

Laksmitawati et al., 2016; 2017; Saanin et al., 2020;

Widowati et al., 2016; 2021).

2.3 Viability Assay

The cytotoxicity of PCE was determined by the

viability of RAW 264.7 cells using MTS assay

(Promega, G3580). This method is used to assess the

sample concentration that is both safe and non-toxic

Anti-inﬂammatory Activities of Pineapple (Ananas comosus) Core Extract in Lipopolysaccharide-induced RAW264.7 Cell Line

for the next assay. The cells density of 5 × 10

cells

per well were plated in medium (DMEM

supplemented with 10% FBS and 1% antibiotic–

antimycotic in a 96-well plate and incubated for 24

hours at 37

C in a humidified atmosphere incubator

with 5% CO

. The medium was washed and replaced

with 99 μL of fresh medium and 1 μL of PCE in two

concentrations (4, 20, 100 µg/mL), and 1% DMSO,

then the plate was incubated for 24 hours. The

negative control group consisted of cells that had not

been treated. The twenty microlitres of MTS were

added to each well. For 4 hours, the plate was

incubated in 5% CO

at 37

C at an incubator. The

absorbance was quantified at 490 nm using a

microplate reader (Multiskan Go, Thermoscientific)

(Widowati et al., 2016; 2021; Laksmitawati et al.,

2016; 2017; Saanin et al., 2020).

2.4 Pro-inflammatory Activation of

RAW264.7 Cells

The cells were seeded at a density of 5 × 10

cells per

well in a 6 well-plate and incubated for 24 hours at

C in a humidified atmosphere with 5%

. The

DMEM was washed and supplemented

with a 1,600

μL growth medium and 200 μL PCE (4, 20 µg/mL)

after being supplemented with 10% FBS and 1%

antibiotic–antimycotic. After 1-2 hours, 200 μL

lipopolysaccharide (1 μg/mL) from Escherichia coli

(Sigma Aldrich, L2880) was added to the

medium

and incubated for 24 hours at 37

C, 5% CO

The

medium was then taken for PGE-2, TNF-α, and IL-1β

levels quantification, centrifuged at 2000 g for 10

minutes, and the supernatant was stored at -80

(Widowati et al., 2016; 2021; Laksmitawati et al.,

2016; 2017; Sandhiutami et al., 2017; Novilla et al.,

2017; Saanin et al., 2020).

2.5 Quantification of PGE-2, TNF-α,

IL-1β Levels in RAW264.7 Cells

The measurement of PGE-2, TNF-α, and IL- 1β levels

were conducted based on the ELISA method, using

PGE-2 ELISA Kit (E-EL-M0052), TNF-α ELISA

Standard Kit (Elabscience, E-EL- M0049), and IL-1β

ELISA Kit (Elabscience, E-EL- M0037), respectively

according to the manufacturer’s instructions. The

inhibition activity was calculated based on the

percentage of a PGE-2, TNF-α, and IL- 1β levels

(Widowati et al., 2016; 2021; Laksmitawati et al.,

2016; 2017; Saanin et al., 2020).

2.6 Statistical Analysis

All data were obtained after doing it in triplicate. The

data were presented as mean ± standard

deviation. The

data were analyzed using ANOVA and

Tukey HSD

Post Hoc Test with p < 0.05 using SPSS software

(version 20.0).

RESULTS AND DISCUSSION

3.1 Viability RAW264.7 Cells

The viability assay was performed to determine

the

safe and nontoxic concentration for the next assay,

which was assessed using the MTS assay (Widowati

et al., 2016; 2021). The viability of RAW264.7 cell

lines has been presented in Table 1.

Table 1: The effect of various concentrations of PCE

towards the viability of RAW264.7 cells.

Sample Viability cells (%)

Negative Control 100.00 ±3.54

PCE 4 µg/mL 95.03 ±2.07

PCE 20 µg/mL 92.94 ±1.04

PCE 100 µg/mL 76.86 ±4.09

*The data was presented as mean ± standard deviation from

3 replications. Different superscript letters in the same

column (a, b, c) showed significant differences among

treatments at p <0.05 (Tukey HSD post hoc test).

The increased concentration was correlated with

increased toxicity (<90% viability cells). Table 1

shows the cytotoxicity of PCE concentration on

RAW264.7 cell lines. The viability of cells was

decreased in a concentration-dependent manner. In

concentration 100 µg/mL demonstrated the lowest

viability of cells by PCE with a value of 76.86 ±

4.09%. Based on the data (Table 1), the safe and un-

toxic of PCE in murine macrophage cells were 4, 20

µg/mL, these concentrations were used for the

treatment in LPS-induced RAW264.7 cells as

inflammation cells model.

3.2 Effect of PCE toward PGE-2 Level

in LPS-induced RAW264.7 Cells

Inhibitory activity of PGE-2 generation from COX-2

also causes an anti-inflammatory effect (Mahesh et

al., 2021). The RAW 264.7 murine macrophage cell

line is commonly used as an in vitro inflammatory

model (Widowati et al., 2016; 2021). The

ICE-TES 2021 - International Conference on Emerging Issues in Technology, Engineering, and Science

inflammatory response is marked releasing of the

PGE-2 level. The anti-inflammatory activity was

showed by decreasing PGE-2 level in LPS-induced

RAW264.7 cells as inflammatory cells model.

*The data was presented as mean ± standard deviation from 3

replications. Negative control: untreated cell; Positive control:

LPS-induced RAW264.7 cells. Different superscript letters (a, b,

c) showed significant differences among treatments at p <0.05

(Tukey HSD post hoc test).

Figure 1: Effects of PCE toward PGE-2 level in LPS-

induced RAW264.7 cells.

PCE 20 µg/mL has a better ability to suppress

PGE-2 level (507.68 pg/mL) compared to PCE 4

µg/mL (795.37 pg/mL) and positive control (LPS-

induced cells) with value 870.48 ± 39.54 pg/mL

(Figure 1). Our findings provide evidence that PCE

significantly inhibits the production of PGE-2

without affecting the cell viability in LPS-induced

RAW 264.7 cells.

A. comosus leaf extract has an ability to inhibit

PGE2 production in a dose-dependent manner with a

value of 2198.83 ± 280.87 pg/mL in the highest

concentration (500 µg/ml). The inhibition of COX-2

enzyme induction and subsequent inhibition of COX-

2 mRNA expression may be the mechanism behind

some phytoconstituents anti-inflammatory activity

(Yatoo et al., 2018). It has also been discovered that

inhibiting COX-2 development results in a lower level

of PGE (Kasemsuk et al., 2018).

The methanol extract of fruit peel (MEFP) of A.

comosus has anti-inflammatory potential through

decreasing the level of PGE-2, so it has the potential

to protect cartilage from damage caused during

rheumatoid arthritis (Kargutkar and Brijesh, 2016).

The anti-inflammatory effect of bromelain also

was correlated with reduced LPS-induced nuclear

factor-kappaB (NF-κB) activity and cyclooxygenase

2 (COX-2) mRNA expression in rat livers (Kasemsuk

et al., 2018). The roles of bromelain are also well

recognized in activating the healthy immune system

with the rapid response to cellular stress (Rathnavelu

et al., 2016).

3.3 Effect of PCE towards TNF-α

Level in LPS-induced RAW264.7

Cells

TNF-α is one of the most important pro- inflammatory

cytokines and is mainly produced by monocytes and

macrophages. These pro- inflammatory cytokines

influence the proliferation and death of cells and it is

secreted during the early phase of acute and chronic

inflammatory diseases such rheumatoid arthritis

(Wang and He, 2018; Saanin et al., 2020).

*The data was presented as mean ± standard deviation from 3

replications. Negative control: untreated cell; Positive control:

LPS-induced RAW264.7 cells. Different superscript letters (a, ab,

bc, c) showed significant differences among treatments at p <0.05

(Tukey HSD post hoc test).

Figure 2: Effects of PCE toward TNF-α level in LPS-

induced RAW264.7 cells.

Treatment with PCE 20 µg/mL potentially inhibit

TNF-α production (345.90 pg/mL) compared to PCE

4 µg/mL (474.19 pg/mL) and also positive control

with value 581.71 pg/mL (Figure 2). This treatment

indicates that PCE had anti-inflammatory activity.

Based on another study, A. comosus extract has a

significant antioxidant activity, as well as anti-

inflammatory ability. It is thought that molecules with

both an anti-oxidative effect and an anti-

inflammatory effect should be effective in treating

diseases caused by oxidative stress as a result of ROS

generation as well as in inflammatory diseases (Lee

et al., 2018; Vrianty et al., 2019).

Kargutkar and Brijesh’s (2017) study showed that

A. comosus leaf extract (ALE) has anti-inflammatory

activity due to significantly decreasing the release of

TNF-α, IL-1β, PGE-2, and ROS by LPS-stimulated

macrophages in a dose-dependent manner. ALE can

decrease the secretion of TNF-α at a concentration of

500 µg/mL with a maximum reduction of 409.89

pg/mL (Kargutkar and Brijesh, 2017).

Anti-inﬂammatory Activities of Pineapple (Ananas comosus) Core Extract in Lipopolysaccharide-induced RAW264.7 Cell Line

The phytoconstituents prevent inflammation and

apoptosis by inhibiting TNF-α in human

macrophages (Wee et al., 2020). Bromelain in A.

comosus potentially inhibits the pro-inflammatory

mediators productions of NFкB, IL-1β, IL-6, TNF-α,

PGE-2 (Bakare et al., 2021). In another study was

reported that Bromelain can inhibit the secretion of

IL-1, IL-6, and TNF by peripheral blood mononuclear

cells (PBMCs) and modulate surface adhesion

molecules on T cells, macrophages, and natural killer

cells (Pavan et al., 2012). Bromelain inhibits the Raf-

1/extracellular-regulated-kinase- (ERK-) 2 pathways

by inhibiting T cell signaling (Kwatra, 2019).

Bromelain also suppressed the LPS-activated

extracellular signal-regulated kinase (ERK), c-Jun N-

terminal kinase (JNK), and p38 mitogen-activated

protein kinase (MAPK) (Insuan et al., 2021).

However, the anti-inflammatory effects of the

bromelain preparation in vitro and in vivo studies

suggest its therapeutic potentials (Kasemsuk et al.,

2018).

3.4 Effect of PCE towards IL-1β

Level in LPS-induced RAW264.7

Cells

IL-1β is a potent immunomodulator that regulates a

variety of immune and inflammatory responses,

including B and T cells activation (Rapsinski et al.,

2015).

PCE 20 µg/mL decreased the IL-1β level (217.63

pg/mL) compared to PCE 4 µg/mL (350.78 pg/mL)

and also positive control with value 433.53 pg/mL.

PCE has anti-inflammatory properties through

inhibitory of IL-1β level among treatments.

Bromelain is a crude, aqueous extract derived

from the stem and fruit of the pineapple plant, which

contains a variety of proteolytic enzymes and has

anti-inflammatory and analgesic properties (Cai et al.,

2017). Bromelain from A. comosus also has ability to

decreases NO, PGE-2, TNF-α, IL-1β and IL-6

production by downregulation of the NF-κB, AP-1,

and JAK/ STAT signaling pathways in LPS-induced

RAW 264.7 macrophages (Lee et al., 2017;

Kargutkar and Brijesh, 2016).

*The data was presented as mean ± standard deviation from 3

replications. Negative control: untreated cell; Positive control:

LPS-induced cell. Different superscript letters (a, b, c, d) showed

significant differences among treatments at p <0.05 (Tukey HSD

post hoc test).

Figure 3: Effects of PCE toward IL-1β level in LPS-

induced RAW264.7 cells.

Bromelain has the ability to modulate the immune

response to reduce the allergic reaction and to

modulate macrophages, NK cells, and T cells. It also

increases the secretion of IL-1β, IL-6, and TNF- α

(Cai et al., 2017). Flavonoids and tannins compounds

in A. comosus can be attributed in anti- inflammatory

properties (Jiang et al., 2014).

4 CONCLUSIONS

PCE has the potential as an anti-inflammatory by

decreasing PGE2, TNF-α, and IL-1β levels. However,

PCE protects against LPS-induced RAW264.7 cells

via inhibiting oxidative stress and inflammation.

ACKNOWLEDGEMENTS

This research was funded by Aretha Medika Utama

Biomolecular and Biomedical Research Center

(AMU-BBRC), Bandung, Indonesia. The authors

also would like to thank Ervi Afifah and Seila

Arumwardana from AMU-BBRC for their valuable

assistance.

REFERENCES

Bakare, A. O., Owoyele, B. V. (2021). ‘Bromelain reduced

pro-inflammatory mediators as a common pathway that

mediate antinociceptive and anti-anxiety effects in

sciatic nerve ligated Wistar rats’. Sci. Rep. 11(1), 1-13.

Cai, T., Verze, P., La Rocca, R., Palmieri, A., Tiscione, D.,

Luciani, L. G., Mazzoli, V., Malossini, G. (2017). ‘The

clinical efficacy of pollen extract and vitamins on

ICE-TES 2021 - International Conference on Emerging Issues in Technology, Engineering, and Science

chronic prostatitis/chronic pelvic pain syndrome is

linked to a decrease in the pro-inflammatory cytokine

interleukin-8’. World J. Mens Health. 35(2), 120- 128.

Chen, L., Deng, H., Cui, H., Fang, J., Zuo, Z., Deng,

J., Li, Y., Wang, X., Zhao, L. (2018).

‘Inflammatory responses and inflammation- associated

diseases in organs’. Oncotarget, 9(6), 7204.

Insuan, O., Janchai, P., Thongchuai, B., Chaiwongsa, R.,

Khamchun, S., Saoin, S., Insuan, W., Pothacharoen, P.,

Apiwatanapiwat, W., Boondaeng, A., Vaithanomsat, P.

(2021). ‘Anti- Inflammatory effect of pineapple

rhizome bromelain through downregulation of the

NFκB- and MAPKs-signaling pathways in

Lipopolysaccharide (LPS)-stimulated RAW264. 7

cells’. Curr. Issues Mol. Biol. 43(1), 93-106.

Jiang, J., Yuan, X., Wang, T., Chen, H., Zhao, H., Yan,

X., Wang, Z., Sun, X., Zheng, Q. (2014).

‘Antioxidative and cardioprotective effects of total

flavonoids extracted from Dracocephalum moldavica

L. against acute ischemia/reperfusion- induced

myocardial injury in isolated rat heart’. Cardiovasc.

Toxicol. 14(1), 74-82.

Kargutkar, S., Brijesh, S. (2016). ‘Anti-rheumatic activity

of Ananas comosus fruit peel extract in a complete

Freund’s adjuvant rat model’. Pharm. Biol. 54(11),

2616-2622.

Kargutkar, S., Brijesh, S. (2017). ‘Anti-inflammatory

evaluation and characterization of leaf extract of

Ananas comosus’. Inflammopharmacol. 26(2), 469-

477.

Kasemsuk, T., Vivithanaporn, P., Unchern, S. (2018). ‘Anti-

inflammatory effects of bromelain in LPS- induced

human U937 macrophages’. Chiang Mai J. Sci. 45(1),

299-307.

Kwatra, B. (2019). ‘A review on potential properties and

therapeutic applications of bromelain’. World J. Pharm.

Pharm. Sci, 8, 488-500.

Laksmitawati, D. R., Prasanti, A. P., Larasinta, N., Syauta,

G. A., Hilda, R., Ramadaniati, H. U., Widyastuti, A.,

Karami, N., Afni, M., Rihibiha, D. D., Kusuma, H. S.

W., Widowati, W. (2016). ‘Anti-Inflammatory potential

of gandarusa (Gendarussa vulgaris Nees) and soursoup

(Annona muricata L) extracts in LPS stimulated-

macrophage cell (RAW264. 7)’. J. Nat. Remed.

16(2), 73-81.

Laksmitawati, D. R., Widyastuti, A., Karami, N., Afifah,

E., Rihibiha, D. D., Nufus, H., Widowati, W. (2017).

‘Anti-inflammatory effects of Anredera cordifolia and

Piper crocatum extracts on lipopolysaccharide-

stimulated macrophage cell line’. Bangladesh J.

Pharmacol. 12(1), 35- 40.

Lee, J. H., Lee, J. B., Lee, J. T., Park, H. R., Kim, J. B.

(2018). ‘Medicinal effects of bromelain (Ananas

comosus) targeting oral environment as an anti-oxidant

and anti-inflammatory agent’. J. Food Nutr. Res.

6(2018), 773-784.

Lee, S. B., Lee, W. S., Shin, J. S., Jang, D. S., Lee, K. T.

(2017). ‘Xanthotoxin suppresses LPS-induced

expression of iNOS, COX-2, TNF-α, and IL-6 via AP-1,

NF-κB, and JAK-STAT inactivation in RAW 264.7

macrophages’. Int. Immunopharmacol. 49, 21-29.

Mahesh, G., Kumar, K. A., Reddanna, P. (2021). ‘Overview

on the discovery and development of anti-inflammatory

drugs: Should the focus be on synthesis or degradation

of PGE2?’. J. Inflamm. Res. 14, 253-263.

Novilla, A., Djamhuri, D. S., Nurhayati, B., Rihibiha, D. D.,

Afifah, E., Widowati, W. (2017). ‘Anti- inflammatory

properties of oolong tea (Camellia sinensis) ethanol

extract and epigallocatechin gallate in LPS-induced

RAW 264.7 cells’. Asian Pac. J. Trop. Biomed. 7(11),

1005-1009.

Pavan, R., Jain, S., Kumar, A. (2012). ‘Properties and

therapeutic application of bromelain: a review’.

Biotechnol. Res. Int., 1-6.

Rahman, I. A., Mohamed, E., Camalxaman, S. N., Haron,

N., Rambely, A. S. (2020). ‘Ananas comosus (L.)

Merr.: A mini review of its therapeutic properties:

Medicinal benefits of pineapple plant’. Healthscope:

The Official Research Book of Faculty of Health

Sciences, UiTM, 3(2), 54-59.

Ramli, A. N. M., Manas, N. H. A., Hamid, A. A. A., Hamid,

H. A., Illias, R. M. (2018). ‘Comparative structural

analysis of fruit and stem bromelain from Ananas

comosus’. Food Chem. 266, 183-191.

Rathnavelu, V., Alitheen, N. B., Sohila, S., Kanagesan, S.,

Ramesh, R. (2016). ‘Potential role of bromelain in

clinical and therapeutic applications’. Biomed. Rep.

5(3), 283-288.

Rapsinski, G. J., Wynosky-Dolfi, M. A., Oppong, G. O.,

Tursi, S.A., Wilson, R. P., Brodsky, I. E., Tukel, C.

(2015). ‘Toll-like receptor 2 and NLRP3 cooperate to

recognize a functional bacterial amyloid, curli’. Infect.

Immun. 83, 693- 701.

Saanin, S. N., Wahyudianingsih, R., Afni, M., Afifah, E.,

Maesaroh, M., Widowati, W. (2020). ‘Suppression of

pro-inflammatory cytokines and mediators production

by ginger (Zingiber officinale) ethanolic extract and

gingerol in lipopolysaccharide-induced RAW264. 7

murine macrophage cells’. Indian J Nat. Prod.

Resour. 11(4), 260-266.

Sandhiutami, N. M. D., Moordiani, M., Laksmitawati, D.

R., Fauziah, N., Maesaroh, M., Widowati, W. (2017).

‘In vitro assesment of anti- inflammatory activities of

coumarin and Indonesian cassia extract in RAW264. 7

murine macrophage cell line’. Iran. J. Basic. Med.

Sci. 20(1), 99-106.

Vrianty, D., Qodariah, R. L., Widowati, W., Sinaga, A. P.

F., Fibrina, D., Fachrial, E. (2019).‘Comparison of

antioxidant and anti-tyrosinase activities of pineapple

(Ananas comosus) core extract and luteolin compound’.

JKB. 30(4), 240- 246.

Wang, T., He, C. (2018). ‘Pro-inflammatory cytokines:

The link between obesity and

osteoarthritis’. Cytokine Growth F. R., 44, 38-50.

Wee, H. N., Neo, S. Y., Singh, D., Yew, H. C., Qiu,Z. Y.,

Tsai, X. R. C., How, S. Y., Caleb Yip, K.

Y., Tan, C. H.,

Koh, H. L. (2020). ‘Effects of

Vitex

trifolia production in

human U937 macrophages’. BMC Complement. Altern.

Anti-inﬂammatory Activities of Pineapple (Ananas comosus) Core Extract in Lipopolysaccharide-induced RAW264.7 Cell Line

Med. Ther. 20(1), 1-15. L. leaf extracts and

phytoconstituents on cytokine

Widowati, W., Darsono, L., Suherman, J., Fauziah, N.,

Maesaroh, M., Erawijantari, P. P. (2016). ‘Anti-

inflammatory effect of mangosteen (Garcinia

mangostana L.) peel extract and its compounds in LPS-

induced RAW264. 7 cells’. Nat. Prod. Sci. 22(3),

147-153.

Widowati, W., Murti, H., Widyastuti, H., Laksmitawati, D.

R., Rizal, R., Kusuma, H. S. W., Sumitro, S. B., Widodo,

M. A., Bachtiar, I. (2021b). ‘Decreased inhibition of

proliferation and induction of apoptosis in breast cancer

cell lines (T47D and MCF7) from treatment with

conditioned medium derived from hypoxia- treated

Wharton’s jelly MSCs compared with normoxia-

treated MSCs’. Int. J. Hematol. Oncol. Stem Cell Res.

15(2), 77-89.

Widyanto, R. M., Nafi'Halimah, R., Rahmi, Y., Utomo, B.,

Proborini, W. D., Yunimar, Y. (2020). ‘Antioxidant

and cytotoxic effect of water

extract of

Ananas comosus

in human breast cancer

cell line’. J. Islamic Med. 4(2),

123-130.

Yatoo, M., Gopalakrishnan, A., Saxena, A., Parray, O. R.,

Tufani, N. A., Chakraborty, S., Tiwari, R., Dhama, K.,

Iqbal, H. (2018). ‘Anti-inflammatory drugs and herbs

with special emphasis on herbal medicines for

countering inflammatory diseases and disorders-a

review’. Recent. Pat. Inflamm. Allergy Drug. Discov.

12(1), 39-58.

ICE-TES 2021 - International Conference on Emerging Issues in Technology, Engineering, and Science