NUS ANT AR A BIOS C IENC E
Vol.
No.
2, pp.
ISSN: 2087-3948
E-ISSN: 2087-3956
DOI: 10.
13057/nusbiosci/n100207 May 2018 Tracheospasmolytic and anti-inflammatory activity of indigenous Curcuma species as traditional antiasthmatic medicines SWANDARI PARAMITA1,2,ou.
EMIL BACHTIAR MOERAD3.
SJARIF ISMAIL2,4.
EVA MARLIANA5 Department of Community Medicine.
Faculty of Medicine.
Universitas Mulawarman.
Jl.
Kerayan.
Kampus Gunung Kelua.
Samarinda 75119.
East Kalimantan.
Indonesia.
Tel.
/fax.
: 62-541-748581.
Cemail: swandariparamita@gmail.
Center for Medicine and Public Health Research.
Institute for Research and Community Services.
Universitas Mulawarman.
Jl.
Kerayan No.
1 Gedung A 20.
Kampus Gunung Kelua.
Samarinda 75119.
East Kalimantan.
Indonesia Department of Internal Medicine.
Faculty of Medicine.
Universitas Mulawarman.
Jl.
Kerayan.
Kampus Gunung Kelua.
Samarinda 75119.
East Kalimantan.
Indonesia Department of Pharmacology.
Faculty of Medicine.
Universitas Mulawarman.
Jl.
Kerayan.
Kampus Gunung Kelua.
Samarinda 75119.
East Kalimantan.
Indonesia Department of Chemistry.
Faculty of Mathematics and Natural Sciences.
Universitas Mulawarman.
Jl.
Barong Tongkok No.
Kampus Gunung Kelua.
Samarinda 75123.
East Kalimantan.
Indonesia Manuscript received: 2 April 2018.
Revision accepted: 19 May 2018.
Abstract.
Paramita S.
Moerad EB.
Ismail S.
Marliana E.
Tracheospasmolytic and anti-inflammatory activity of indigenous Curcuma species as traditional antiasthmatic medicines.
Nusantara Bioscience 10: 105-110.
Asthma is still a major health problem in the world.
Antiasthma drugs raise the problem of side effects and medical expenses.
Thus, it is necessary to develop medicinal plants based antiasthma drugs with fewer side effects and more affordable costs.
The potential antiasthma medicinal plant came from the genus of Curcuma (Zingiberaceae famil.
Turmeric (Curcuma longa L.
) is the most prominent species in this genus and has been extensively studied for the treatment of asthma.
However, there are other Curcuma indigenous species that are also traditionally used for asthma, namely Curcuma aeruginosa Roxb.
and Curcuma mangga Val.
Zijp.
Both indigenous species have not been studied pharmacological activity as an antiasthma.
This research was conducted to know the tracheospasmolytic activity of ethanol extract of C.
aeruginosa and mangga on the separate organ of guinea pig trachea to determine the antiasthma effect.
The research was also conducted to know anti-inflammatory activity with membrane stabilization test and inhibition of lipoxygenase.
Results showed that C.
aeruginosa and C.
mangga had lower tracheospasmolytic activity when they were compared with aminophylline as a positive control.
However, the EC50 value of C.
020 A 0.
and C.
039 A 0.
were not significantly different with aminophylline .
as positive controls.
The result of membrane stabilization test showed that the EC50 value of C.
76 A 1.
and C.
50 A 0.
were high when they were compared with indomethacin .
39 A 2.
as the positive control.
Results of lipoxygenase inhibition test showed that the EC50 value of C.
aeruginosa (-111.
11 A 2.
and C.
mangga (-101.
98 A 1.
were high than that of indomethacin (-282.
84 A 7.
as the positive control.
These results show the potential use of C.
aeruginosa and C.
as antiasthma via tracheospasmolytic activity in separate organs of guinea pig.
They also had anti-inflammatory effect based on membrane stabilization and inhibition of lipoxygenase test.
Keywords: Anti-asthma, anti-inflammatory.
Curcuma aeruginosa.
Curcuma mangga, tracheospasmolytic INTRODUCTION Asthma is a chronic inflammatory airway disease characterized by the occurrence of respiratory airway hyper response and reversible narrow airway (Kasper et al.
Asthma is one of the major non-communicable diseases in the world.
About 235 million people worldwide suffer from asthma, especially in children.
The main risk factor for the occurrence of asthma is a combination of genetic susceptibility to environmental exposures such as air pollution (WHO 2.
Asthma medications are given to manage asthma sufferers (Papadakis and McPhee 2.
Herbal preparations are one of the most popular complementary treatment used by asthmatic patients.
Almost 40% of people with asthma use herbal medicines.
Many important asthma drugs such as B2-agonists, anticholinergics, methylxanthines, and cromones have herbal origins (Karaman et al.
Research also shows that some medicinal plants have the effect of reducing smooth muscle stiffness.
The mechanism of those medicinal plants is similar to the mechanism of asthma drugs, especially the anticholinergic drug (Amit and Vandana 2.
Research also shows that some medicinal plants have the anti-inflammatory effect, which is the same with the mechanism of corticosteroid drug in asthma drugs (Paramita et al.
The genus Curcuma (Family Zingiberacea.
comprising more than 100 species have been recognized widely as food and traditional medicines.
is the most studied species of all Curcuma species (Akarchariya et al.
Indonesia is home to many species of Curcuma and several species of these have been commonly used as traditional medicines.
The various species of Curcuma often used are C.
mangga, and Turmeric appears to be the species, which is most frequently used as raw materials for traditional medicine in Indonesia, followed by C.
The N U S A N T A R A B I O S C I E N C E 10 .
: 105-110.
May 2018 other species, including C.
aeruginosa and C.
mangga, are considered as indigenous Curcuma species, which are not intensively studied, yet (Rahayu et al.
The rhizomes of other Curcuma species are also pharmacologically important but several of these species are not exploited commercially (Shakeri et al.
Important medicinal plants from genus Curcuma having anti-asthmatic potential is C.
Other rhizomes of Curcuma species are traditionally used in the treatment of asthma, i.
aeruginosa (Rajamma et al.
mangga (Dewangan et al.
caesia (Pathan et al.
Pakkirisamy et al.
, and C.
zedoaria (Pathan et The examination of antiasthma effects of two indigenous Curcuma species, i.
aeruginosa, and C.
Mangga is not studied, yet.
Therefore, the objective of this study was to know the tracheospasmolytic and antiinflammatory activity of C.
aeruginosa and C.
applied on isolated trachea of guinea pig.
MATERIALS AND METHODS
Sampling and sample identification The sampling of medicinal plants was conducted at the Kutai Kartanegara District.
East Kalimantan.
Plants were then identified in the Department of Biology.
Faculty of Mathematics and Natural Sciences.
Mulawarman University to certify the plant legality.
This study has obtained the ethical clearance of the Medical and Health Research Ethics Commission.
Faculty of Medicine.
Mulawarman University.
This study was conducted in the Pharmacology Laboratory of the Faculty of Medicine.
Mulawarman University.
Plant extractions The extraction of medicinal plant followed the guidelines from the Indonesian Herbal Pharmacopoeia (MoH RI 2.
The rhizomes of C.
aeruginosa and C.
mangga were dried and then crushed into simplicia.
The simplicia were then macerated using absolute ethanol solvent in a ratio of 1 part of simplicia to 10 parts of The mixture was soaked for 6 hours followed by stirring occasionally with an orbital shaker at room temperature in each interval hours, which were then stood for 18 hours.
The mixture was separated using filter paper, followed by the evaporation using a rotary evaporator at 50AC.
The obtained viscous extract was dried to obtain a dry extract.
The dried extracts were then stored in a refrigerator of -20AC for further study.
Tracheospasmolytic activity Tracheospasmolytic activity of medicinal plant extracts was tested in separated organs of tracheal guinea pigs via histamine injection.
Guinea pigs were sacrificed and the trachea was quickly dissected by adhering fat and connective tissue.
Trachea was placed in the petri dish containing Krebs Physiological Salt Solution (PSS).
The rings were suspended in L-shaped wire loops in 10 ml organ baths containing the Krebs PSS aerated with carbogen by maintaining the temperature at 37AC.
Isometric contractions of tracheal rings were measured by the transducer (Force Transducer.
coupled to the amplifier (BridgeAm.
connected to PC running ChartV5 software, all from ADInstruments (Dunedin.
New Zealan.
equilibration period of 90 minutes was allowed in Krebs PSS, at the end of the period, the tracheal rings were stimulated with histamine in order to establish viability.
After equilibration, the tracheal rings were exposed to aminophylline as positive control drugs and medicinal plants extract of C.
aeruginosa and C.
mangga according to the experimental protocol.
In each preparation, a single concentration-relaxation curve was obtained.
The relaxant effect induced by positive control drug and medicinal plant extracts was expressed as the reverse percentage of initial contraction force, which was elicited by histamine in tracheal rings (Ozolua et al.
Janbaz et al.
Lima et al.
Figure 1.
Rhizome of Curcuma aeruginosa Roxb.
Rhizome of Curcuma mangga Val.
Zijp PARAMITA et al.
Ae Tracheospasmolytic and anti-inflammatory activity of Curcuma Anti-inflammatory activity Anti-inflammatory activity of medicinal plant extracts was measured by membrane stabilization and lipoxygenase inhibition test.
For membrane stabilization test, blood samples were collected into an anticoagulant and centrifuged at 3000 rpm for 10 minutes at room The supernatants .
lasma and leucocyte.
were carefully removed while the packed red blood cell was washed in fresh normal saline.
The process of washing and centrifugation were repeated five times until supernatants were clear.
The membrane stabilizing activity assay was carried out using erythrocyte suspension while Indomethacin was used as drug standard.
The mixtures consisted of hyposaline sodium chloride, sodium phosphate buffer, erythrocyte suspension, drug standard, plant extracts and final reaction mixtures were mixed to produce Drugs were mixed in the blood control, while the drug control did not contain the blood suspension.
The reaction mixtures were incubated at 56AC for 30 min on a water bath.
The tube was cooled under running water followed by centrifugation at 5000 rpm for 10 min at room The supernatant was collected.
The absorbance of the released hemoglobin from the supernatant was measured at 560 nm (Omale and Okafor Oyedapo et al.
For lipoxygenase inhibition test, a mixture of a solution of sodium borate buffer and lipoxygenase enzyme was incubated with plant extract sample at room temperature for 5 minutes.
The reaction was started by the addition of linolic acid substrate.
The absorbance of the resulting mixture was measured at 234 nm as a function of time at a rate of one measurement/minutes .
Indomethacin was used as a positive standard (Khasawneh et al.
Singh et al.
Data analysis Tracheheospasmolytic activity is tabulated in the mean A SE curve of the dose-response.
The value of EC50 was The homogeneity and variance analysis was conducted when the data showed the normal distribution and variance followed by t-test analysis.
The significant difference was performed when p <0.
Membrane stabilization activity and lipoxygenase inhibition were tabulated in mean A SD form, in order to calculate EC50 The difference between the control was performed based on t-test analysis with the significance of p <0.
00 A 0.
, .
85 A 0.
, .
80 A 0.
whereas in aminophylline obtained results .
82 A 1.
, .
68 A 1.
(-3.
94 A 3.
, (-23.
78 A 5.
, (-57.
78 A 5.
, (-93.
The result of trachea relaxation between negative control, aminophylline and group with plant extracts is presented in Figure 2.
The result showed that the decrease of spasmolytic activity of C.
aeruginosa and C.
mangga in the guinea pig tracheal tone was significantly better .
<0.
than that in negative control.
The EC50 value of C.
020 A 0.
and C.
039 A 0.
was higher than aminophylline .
016 A 0.
as a positive control.
This shows that the effect of the plant extract was not as strong as aminophylline, as shown in Figure 3.
However, the C.
aeruginosa had an indifferent value from that of aminophylline as a positive control.
This suggests that C.
aeruginosa has a better efficacy than C.
mangga as spasmolytic when they were compared with aminophylline.
Figure 2.
Graph of trachea relaxation differences between C.
aeruginosa (CA).
mangga (CM), aminophylline (A) as the positive control and negative control (N) in trachea isolates organ
of guinea pig
RESULTS AND DISCUSSION
Tracheospasmolytic activity The results of spasmolytic activity of ethanol extract of aeruginosa on separated organ of trachea guinea pig at 01, 0.
03, 0.
1, 0.
3, 1 and 3 mg/ml concentrations were as follows: .
90 A 2.
, .
44 A 1.
, .
02 A 3.
, .
, (-0.
48 A 3.
, (-2.
02 A 4.
, .
25 A 2.
, .
13 A 2.
, .
11 A 2.
, .
, .
90 A 2.
and on control .
olvent extrac.
obtained results .
31 A 0.
, .
68 A 0.
, .
54 A 0.
Figure 3.
The EC50 result on tracheospasmolytic activity between aeruginosa (CA).
mangga (CM) and aminophylline (A) as a positive control .
<0.
N U S A N T A R A B I O S C I E N C E 10 .
: 105-110.
May 2018 Anti-inflammatory activity The result of membrane stabilization test showed that the EC50 value of C.
76 A 1.
mg/mL and mangga .
50 A 0.
mg/mL were higher compared with indomethacin .
39 A 2.
mg/mL as the positive control, as shown in Figure 4.
The results of lipoxygenase inhibition test showed that the EC50 value of C.
(-111.
11 A 2.
g/mL and C.
mangga (-101.
98 A 1.
g/mL were also higher than indomethacin (-282.
g/mL as the positive control (Figure .
The smaller EC50 concentration determines the better anti-inflammatory activity based on membrane stabilization test and lipoxygenase inhibition.
The results showed that indomethacin as positive control had better antiinflammatory activity than that in C.
aeruginosa and C.
mangga, based on membrane stabilization test and lipoxygenase inhibition.
Figure 4.
The EC50 results on membrane stability test between C.
aeruginosa (CA) and C.
mangga (CM) with indomethacin (IND) as the positive control .
<0.
Figure 5.
The EC50 results on lipoxygenase inhibition test between C.
aeruginosa (CA) and C.
mangga (CM) with indomethacin (IND) as the positive control .
<0.
Discussion Curcuma longa from genus Curcuma is the most important medicinal plants, which is traditionally used as antiasthma for the treatment of respiratory disorders such as asthma, bronchitis and allergic reactions in the respiratory tract (Gonuguntla et al.
Anti-asthmatic and antioxidant property of C.
longa has been tested in the experimental animal model of asthma (Prasad et al.
Bronchodilatory effect of C.
longa may be due to the mast cell stabilizing activity, suppression of immunoglobulin E, and inhibition of inflammatory mediators (Gohil and Mehta Dongre et al.
Curcuma caesia.
aeruginosa, and C.
mangga are another Curcuma species traditionally used as asthma treatment.
caesia tested antiasthma for its relaxant effect in guinea pig trachea has been successfully reported as powerful protection against histamine-induced bronchospasm (Paliwal et al.
significantly decreased milk induced leukocytosis and showed anti-inflammatory activities (Pathan et al.
In this study, the examination of antiasthma effects of C.
aeruginosa and C.
mangga has been firstly reported.
Curcuma aeruginosa is known in Indonesia as Autemu irengAy and Aupink and blue gingerAy in English (Nurcholis et Simoh and Zainal 2.
aeruginosa has been used as traditional medicine in South and Southeast Asia for gastrointestinal and uterine disorders or parasitic and fungal infection (Jose and Thomas 2014.
Hossain et al.
Theanphong et al.
Sesquiterpenes have been found as common chemical constituents of C.
(Suphrom et al.
Curcuma mangga was known as Autemu manggaAy in Indonesia, meaning mango-like turmeric (Kaewkroek et al.
The rhizomes of C.
mangga are used as traditional treatment for stomach and chest pain, fever, debility, bronchitis, aphrodisiac, postpartum care and cancer-related diseases (Wan-Ibrahim et al.
Malek et al.
The composition of C.
mangga is dominated by sesquiterpenes and monoterpenes (Kamazeri et al.
It is suggested that curcumin with other chemical compounds from C.
aeruginosa and C.
mangga could show antiasthma activity.
Anti-asthmatic and antioxidant property of curcumin has been tested in the experimental animal model of asthma (Houssen et al.
Curcumin is acted to inhibit histamine release (Kale et al.
Curcumin alleviates the pathological changes of chronic asthma, by reducing lung and airway inflammation in asthma induced animal (Chong et al.
Yang et al.
Curcumin improves the airway obstruction by inhibiting mast cell degranulation so that it is safely used as therapy for asthma patients (Subhashini et al.
Abidi et al.
Chauhan et al.
The results of this study indicate that ethanol extract of aeruginosa and C.
mangga has an antiasthma effect based on the tracheospasmolytic and anti-inflammatory Therefore.
aeruginosa and C.
mangga can be developed as a source of the new antiasthma drug.
PARAMITA et al.
Ae Tracheospasmolytic and anti-inflammatory activity of Curcuma
ACKNOWLEDGMENTS
Acknowledgments to the Directorate General for Research and Development.
Ministry of Research and Technology Higher Education of the Republic of Indonesia, for financing this research, as part of the implementation of Basic Research of Flagship University Year of 2018.
REFERENCES