The Effect of Poguntano Leaf Extract (Curanga feel-terrae Merr.)

on Procalcitonin Levels in Acute Bacterial Rhinosinusitis Rat Model

Andrina Yunita Murni Rambe

, Delfitri Munir

, Rosita Juwita Sembiring

, Syafruddin Ilyas

Otolaryngology-Head and Neck Surgery Department, Faculty of Medicine, Universitas Sumatera Utara, Indonesia

Clinical Pathology Department, Faculty of Medicine, Universitas Sumatera Utara, Indonesia

Biology Department, Faculty of Mathematics and Natural Sciences, Universitas Sumatera Utara, Indonesia

Keywords: Poguntano Leaf Extract, Procalcitonin, Acute Bacterial Rhinosinusitis

Abstract: Background: Acute rhinosinusitis is a common condition caused by a viral and bacterial infection.

Procalcitonin (PCT) has been proofed as a biomarker for bacterial infections. PCT levels increase to higher

levels in bacterial infection. The extract of Poguntano leaf contains phytochemicals namely flavonoids,

saponins, tannins, glycosides, terpenoids, and steroids. Objective : This study aimed to determine the effect

of Poguntano leaf extract on Procalcitonin level expression in acute bacterial rhinosinusitis rats model

Methods : Twenty-four female Wistar rats were divided randomly into three groups, negative control (group

A), positive control (group B), group obtaining Poguntano extract 10mg/kg/day orally for 5 days at the 10

day after induction (group C). Rhinosinusitis was induced by the insertion of sponge impregnated with a

solution containing Staphylococcus aureus into nasal cavities of study group B and C. The groups were

euthanized within 1 day (group A), 10 days (group B), and 15 days (group C). Procalcitonin blood level was

measured by ELISA. Results: there was a significant reduction of Procalcitonin levels in the treatment

group than in the control group. Conclusion: this study suggests that Poguntano leaf can reduce

procalcitonin levels in a rat model of acute bacterial rhinosinusitis.

1 INTRODUCTION

Acute rhinosinusitis (ARS) is a common upper

respiratory tract disorder that involves inflammation

of the nasal and paranasal sinus mucosa. It is often

caused by a viral infection of which only 0,5 to 2%

of cases are complicated by a bacterial infection.

However, the efficacy of antibiotics is limited or

controversial. There is significant overuse of

antibiotics due to uncertainty to differentiate

bacterial from viral rhinosinusitis on the basis of

clinical judgment alone.

Compared with non-bacterial ARS, bacterial ARS

encompassed a more powerful local and systemic

inflammatory reaction. These responses were

strongest at the beginning of respiratory symptoms,

indicates that bacterial ARS starts to develop during

an early phase of symptoms. In theory, biomarkers

that parallel the local and systemic inflammatory

reaction could shed light on the pathophysiology and

development of bacterial ARS. (Autio, 2017).

Procalcitonin (PCT), a proinflammatory

biomarker, has been proposed as a pertinent marker

of bacterial infection. It is released ubiquitously in

response to bacterial toxins and bacterial-specific

proinflammatory mediators (Schuetz, 2013; Autio,

2017). Circulating levels of PCT are elevated in

bacterial infections but remain relatively low in viral

infections (Kofteridis, 2009).

Despite the development of new antibiotics and

advances in sinus surgery over the past few decades,

rhinosinusitis is still an enigmatic process.

Herbal medicines have been used for generations

to treat bacterial ARS, and there has been an

increasing interest in herbal medicine both in the

USA and in Europe (Bachert, 2009). Herbal

medicine is a promising alternative in the treatment

of ARS (Koch, 2016).

Poguntano is abundantly growing in Indonesia and

traditionally has been used as stimulant, diuretic,

malaria and recently is used for the treatment of

Diabetes Mellitus (Sitorus, 2014). The extract of

Poguntano leaf contains chemical compounds such

Rambe, A., Munir, D., Sembiring, R. and Ilyas, S.

The Effect of Poguntano Leaf Extract (Curanga feel-terrae Merr.) on Procalcitonin Levels in Acute Bacterial Rhinosinusitis Rat Model.

DOI: 10.5220/0009847800190023

In Proceedings of the 2nd International Conference on Tropical Medicine and Infectious Disease (ICTROMI 2019), pages 19-23

ISBN: 978-989-758-469-5

as flavonoids, saponins, tannins, glycosides,

steroids, and triterpenoids (Patilaya, 2015).

2 MATERIALS AND METHODS

2.1 Preparation of Extract

Poguntano leaves were collected from Pancur Batu,

Deli Serdang District, North Sumatera and

identification of the leaves were performed at

Department of Pharmacology Pharmacy, Faculty of

Pharmacy, University of Sumatera Utara, Indonesia.

2.2 Rats and Groups

Twenty four female Wistar rats aged 6-8 weeks and

weighing 180-200 grams were used. They were

housed in pathogen-free conditions in the Animal

Laboratory of Biology Department, Faculty of

Mathematics and Natural Sciences, University of

Sumatera Utara, Indonesia, and the rats were given

food and water at all times. All animals were kept

under standard conditions for a period of seven days

prior to study initiation. The study was approved by

the Ethics Committee, Medical Faculty, University

of Sumatera Utara.

Rats were randomly divided into three groups of

8. Table 1 shows the interventions for each group.

The rats in group A were sacrificed on the 1

day

without given induction or tested extract (negative

control). The rats in group B were sacrificed 10 days

after the experiment induction (Cheng, 2009; Ozcan,

2011) without given tested extract (positive control)

and the rats in group C were sacrificed 15 days after

the experiment induction with a given extract of

Poguntano leaves 10 mg/BB/day for 5 days orally

(Juwita, 2009) started at the 10

days after

induction. The animals were sacrificed by cervix

dislocation.

Table 1: Group of treatments

Group Treatment

Animals were given standard food and

water ad libitum without induction and

tested extract (negative control)

Animals were given standard food and

water ad libitum with induction but

without tested extract (positive control)

Animals were given standard food and

water ad libitum with induction and extract

of Poguntano leaves 10 mg/BB/day for 5

days orally at the 10

day after induction

2.3 Preparation of Bacterial

Suspension and Induction of

Rhinosinusitis

Strain ATCC 25923 of S aureus was suspended to

reach a concentration of 900x10

cells/ml using a

McFarland Nephelometer Standard III at the

Department of Microbiology, University of

Sumatera Utara Hospital. After the administration of

sedative, the right nasal cavities of the animals were

filled with the sponge for indirect obstruction of the

maxillary sinus ostium (Figure 1).

The sponges were soaked in 1.0 ml solution

containing bacteria. No procedure was performed in

the left nasal cavity.

Figure 1. Insertion sponge into the right nasal cavity

2.4 Procalcitonin Determination

PCT levels were measured in sera using Rat

Procalcitonin (PCT) ELISA kit by Cusabio (CSB-

E13419r) according to the manufacturer’s

instruction with detection range 31.25-2000 pg/ml.

2.5 Data Analysis

All data are expressed as group mean ± SD and

analyzed using SPSS. Kolmogorov-Smirnov test

was applied for the analysis of data distribution. The

parametric test of analysis of variance (ANOVA)

was used to analyze the data. The effects of

treatment were examined using two-way analysis of

ICTROMI 2019 - The 2nd International Conference on Tropical Medicine and Infectious Disease

variance, with a Bonferroni correction for post-hoc

pairwise comparisons. A critical value for

significance of p < 0.05 was used throughout the

study.

3 RESULTS

The results of the phytochemical screening are

presented in Table 2. As can be seen in Table 2, it is

shown that the chemical compounds contained in

Poguntano leave consisted of tannins, saponins,

triterpenoids/steroids, flavonoids, and glycosides.

Table 2: The result of phytochemical screening

No Screening Results

1 Alkaloids Negative

2 Tannins Positive

3 Saponins Positive

4 Triterpenoids/

Steroids

Positive

5 Flavonoids Positive

6 Glycosides Positive

Table 3 : Procalcitonin levels

Group Procalcitonin

levels, Mean

(SD), pg/ml

p B C

A 287,5

(120,62)

<0,001 <0,001 1,000

B 699,38

(214,17)

<0,001

C 191,13

(51,75)

The lowest level of procalcitonin was found in

group C with an average of 191.13 pg/ml (SD =

51.75 pg/ml). While the highest PCT levels were

found in group B (positive control) with an average

value of 699.38 pg/ml (SD = 214.17 pg/ml).

Using the ANOVA test showed that there were

significant differences in the mean PCT in the 3

study groups. From the results of the Post Hoc test

with the Bonferroni test showed that there were

significant differences between groups A and B (p

<0.001) and group B and C (p <0.001). There was

no difference in mean PCT levels between group A

and C (p = 1,000).

4 DISCUSSION

In this study, the highest increase in procalcitonin

(PCT) levels occurred in the ARS-induced group

without given poguntano extract (group B) with an

average value of 699.38 pg/ml (SD = 214.17 pg/ml).

This is in accordance with the statement of Autio

(2017) which states that biomarkers that are

compatible with both local and systemic

inflammatory reactions can show pathophysiological

and bacterial development in ARS, where PCT is

one of the proinflammatory markers that can be

associated with bacterial infections.

PCT is considered the most promising marker

in the assessment of upper respiratory tract

infections to distinguish etiologies from bacterial

and non-bacterial (Kofteridis, et al., 2009). PCT

release is mediated by cytokines which are increased

due to the response of bacterial infections, such as

tumor necrosis factor



and interleukin 1



, while

PCT is retained by the release of interferon due to its

response to viral infection (Dilger, et al., 2019).

The increase in PCT in this study is in

accordance with a series of clinical studies

documenting an increase in PCT levels in shock

septic patients due to bacteremia (Gilbert, 2011). A

cohort study in Wegener granulomatosis also

showed an increase in PCT levels in patients

suffering from bacterial infections, but PCT levels

appear normal in patients with active autoimmune

(Limper, et al., 2010).

Based on the ANOVA test, there was a

significant difference between the groups that were

only induced by ARS (group B) and the group given

poguntano extract (group C).

If assessed from the average, in this study it was

also seen that the lowest PCT levels were found in

the group given poguntano leaf extract ten days after

induction, in group C. Decreased PCT levels after

The Effect of Poguntano Leaf Extract (Curanga feel-terrae Merr.) on Procalcitonin Levels in Acute Bacterial Rhinosinusitis Rat Model

being given this poguntano extract in reducing

inflammatory reaction due to ARS induction

according to the statement Auliafendri, et al. (2019)

which states that modern pharmacological

investigations have indicated that poguntano extract

has an anti-inflammatory, antipyretic, analgesic,

antioxidant, diuretic, antidiabetic, hepatoprotective,

and antihelmintic effect. This is in accordance with

Sibagariang (2017) who concluded from some of the

results of previous studies that poguntano contains

chemical compounds in the class of glycosides,

flavonoids, saponins, tannins and

steroids/triterpenoids. Tannin is considered to have

an anti-inflammatory and anti-diabetes effect. The

activity of tannin as an anti-inflammatory has been

widely investigated (Park, et al, 2013).

In addition, flavonoids (also called

bioflavonoids), are one of the natural ingredients

that can be found in plants, have been considered to

have an anti-inflammatory function both in vitro and

in vivo. Various studies have shown that flavonoids

react through a variety of mechanisms to prevent the

occurrence of inflammatory responses and also areas

cardioprotective, neuroprotective and

chemopreventive agents (Hassan, et al., 2012).

The inflammatory response occurs through three

important phases where each phase is mediated by a

different mechanism, where the acute phase is

characterized by local vasodilation and increased

capillary permeability, the subacute phase is

characterized by infiltration of leukocytes and

phagocytic cells, whereas in the chronic proliferative

phase is indicated by tissue degeneration and fibrosis

(Yassine, et al., 2016).

Yassine, et al., (2016) in his study has demonstrated

the role of flavonoids in the anti-inflammatory

process. In the initial phase where histamine and

serotonin release is followed by edema due to the

production of bradykinin and prostaglandin (this

phase is known to be sensitive to both steroid and

non-steroidal anti-inflammatory agents), flavonoids

are seen to inhibit important enzymes that play a role

in biosynthesis of tissue activator production,

especially prostaglandins and nitric oxide.

Flavonoids are also seen to be effective in inhibiting

arachidonic acid metabolism which mediates

prostaglandin biosynthesis through inhibition of

cyclooxygenase 1 (COX-) and cyclooxygenase 2

(COX-2) enzyme gene expression. The active

ingredient of flavonoids is also considered capable

of being an effective scavenger of reactive oxygen

species due to the presence of phenolic hydroxyl

groups so that it is also considered a strong

antioxidant (Satria, et al., 2017).

5 CONCLUSION

According to our study, it can be concluded that the

extract of Poguntano leaf can reduce procalcitonin

levels in a rat model of acute bacterial rhinosinusitis.

Further investigation is needed to isolate and

identify the compounds present in the extract, which

are responsible for this activity.

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The Effect of Poguntano Leaf Extract (Curanga feel-terrae Merr.) on Procalcitonin Levels in Acute Bacterial Rhinosinusitis Rat Model