EFFECT OF BITTER MELON (Momordica charantia) LEAVES JUICE ON IMMUNOGLOBULIN M (IgM) AND IMMUNOGLOBULIN G (IgG) ACTIVITY OF MALE MICE (MUS MUSCULUS)
Arie Arizandi K., Hajar Astuti, Rangga Meidianto A., Mufidah
(Faculty of Pharmacy, Hasanuddin University, Makassar)
Abstract
Bitter melon (Momordica charantia) is widely growth in tropical area, and the leaves was useful in medicinal. Research about the effect of bitter melon leaves juice for immunomodulatory activity had been done. Three different concentrations, 25% w/v, 50% w/v, and 75% w/v of bitter melon leaves juice in water, with 1 ml/30 g body weight given orally once a day for six days. Then each animal was immunized 1 ml/ 30 g body weight of 2 % Sheep Red Blood Cells (RBC) as the antigenic challenges. Evaluation of Ig activity was conducted at the day 6th and 11th after immunization for IgM and IgG, respectively, using haemagglutinating antibody titter (HAT) method. The result of study indicated that juice of bitter melon leaves increase immunoglobulin M (IgM) four grade higher than negative control, while for the immunoglobulin G (IgG) are twice higher than negative control; and these improvement were statistically significant (P>0.01). The highest Ig activity was achieved by 25% w/v of bitter melon leaves juice.
Keywords : Bitter melon leaves, haemagglutinating antibody titter (HAT), immunoglobulin M (IgM), immunoglobulin G (IgG), sheep Red Blood Cells (RBC).
Introduction
Nowadays, one of the developed principal methods of medicinal that using traditional medicine is by increased the immunity system. If the disease include in infection disease, so the immunity system killed the causes of the disease by indirectly mechanism, that is increased the cellular defender. It is the reason of increasing of immunity system of the patient (Winarno et al.2000).
The defender of body is related with the presence of antibody. Actually, antibody is immunoglobulin protein that being secreted by B cell which is fixated by antigen. Immunoglobulin is the first substances that being identified as molecule in serum that can neutralized the amount of microorganism that caused infection disease. Immunoglobulin M (IgM) is the antibody that very efficient in complement activation, while immunoglobulin G (IgG) can help the phagocitocyt at the destruction of antigen (Bratawidjaja. 2004).
Momordica charantia is a tropical and subtropical vine of the family Cucurbitaceae, widely grown for edible fruit, which is among the most bitter of all vegetables. English names for the plant and its fruit include bitter melon or bitter gourd. (Abascal et al.,2005).
Bitter melons have been used in various Asian traditional medicine systems for a long time. Like most bitter-tasting foods, bitter melon stimulates digestion. While this can be helpful in people with sluggish digestion, dyspepsia, and constipation, it can sometimes make heartburn and ulcers worse. The fact that bitter melon is also a demulcent and at least mild inflammation modulator, however, means that it rarely does have these negative effects, based on clinical experience and traditional reports. (Abascal et al.,2005).
Also known as Ku gua, the herbaceous, tendril-bearing vine grows to 5 m. It bears simple, alternate leaves 4-12 cm across, with 3-7 deeply separated lobes. Each plant bears separate yellow male and female flowers. The young shoots and leaves may also be eaten as greens; in the Philippines, where bitter melon leaves are most commonly consumed, they are called dahon (leaves) ng ampalaya. The seeds can also be eaten, and give off a sweet taste, but have been known to cause vomiting and stomach upset. (Abascal et al.,2005).
Materials and Methods
Materials
Momordica charantia fresh leaves were collected in month of January from Makassar, Indonesia. The fresh leaves were diluted to made the juice with concentration 25% w/v, 50% w/v, and 75% w/v.
Animals
Male mice (20-30 g) were used, animals were housed under standard conditions of temperature (23°C±1°C). Fresh Sheep Red Blood Cell (SRBC) in alsevers solution was obtained from Makassar veterinary.
Antigen
Sheep Red Blood Cell (SRBC) collected in alsevers solution, washed three times in large volumes of pyrogen free 0.9% normal saline and adjusted to a concentration 2% for immunization and challenge.
Treatment
Bitter melon (Momordica charantia) leaves prepared in laboratory in a simple mortar using pestle; animals were divided into four group consisting mice served as control (Group I). the herbal formulation was fed orally at concentration 25% w/v/day (group 2), 50% w/v/day (group 3), and 75% w/v/day (group 4) for assessment immunomodulatory effect.
Haemagglutinating Antibody Titer (HAT)
Mice of group II until IV were pretreated with bitter melon leaves juice for six days and each mouse was immunized with 1ml/30g weight SRBC 2%/mouse by i.p. route. The day of immunization was referred as day 0 after treated with bitter melon leaves juice. After 5 days for IgM and 10 days for IgG, blood sample were collected from each mouse on day 6 for HAT. The titer was determined by tittering serum dilution with SRBC (0.05x109). The micro plates were incubated at 37oC for one hour and room temperature for 24 hours and examined visually for agglutination. The reciprocal of highest dilution of serum showing 50% agglutination has been expressed as HAT.
Statistical Analysis
The data were analyzed using Complete Random Device (CRD) method and then continued with Duncan’s Multiple Range Test (DMRT) method. p>0,05 were considered significant for both of treatment (IgM and IgG).
Results and Discussion
Results
Bitter melon (Momordica charantia) leaves juice was evaluated for Immunoglobulin M (IgM) and immunoglobulin G (IgG) activity. This showed a significant increased in Immunoglobulin M (IgM) and immunoglobulin G (IgG) activity (p>0,05) in mice.
The treatment induced marked enhancement of humoral response in animals. From the study it may be inferred that bitter melon (Momordica charantia) leaves juice promotes increased in Immunoglobulin M (IgM) and immunoglobulin G (IgG) activity and thus rationalizing its traditional claim.
Discussion
Immunomodulatory agents e.g IgM and IgG of the animal origin enhanced against a pathogen by activating the immune system. In the present study Momordica charantia when orally administrated, significantly increased the activity of IgM and IgG.
The data of immunoglobulin G (IgG) and immunoglobulin M (IgM) activity test after given the juice of bitter melon leaves depend on immunoglobulin titter towards to the mice after immunized of sheep Red Blood Cells (RBC) 2% are described on the table below :
Table the data of immunoglobulin M (IgM) activity.
| Treatment | Titer Immunoglobulin G (IgG) | |||
| Control | Cons. 25% | Cons. 50% | Cons. 75% | |
| IgM | 0.81 | 3.21 | 2.01 | 1.61 |
| IgG | 0.81 | 2.01 | 1.61 | 1.41 |
The activity of IgM in concentration 25% w/v was significantly increased when compared to untreated control.
The titer show little bit significant increased change with concentration 50% w/v and 75% w/v of bitter melon (Momordica charantia) leaves juice administration. However, a significant increase was observed at the concentration 25% w/v/day with 4 fold increase compared to control unrated animals (p>0,05). The augmentation of the humoral response as evidence by an enhancement of antibody responsiveness to SRBC in mice as consequently of pre and post immunization treatment indicates the enhanced responsiveness of b-lymphocytes subsets involve antibody synthesis. During Cell Mediated Immunity (CMI) response, sensitized T-lymphocytes, when challenged by the antigen, are converted to lymphoblast and secretes lymphokines, attracting more scavenger cells to the site of reaction. The infiltrating cells are thus immobilized to promote defensive. In our studies, foot volume was enhanced after bitter melon (Momordica charantia) leaves juice treatment suggests cell mediated enhancement (Sen et al.,1992).
Increase in HAT response indicated bitter melon (Momordica charantia) leaves juice potentiates humoral as well as cellular immunity. One of explanations for warded to justify the beneficial effects of indigenous drugs in diseases states is the non specific enhancement of immune states of the organism (Patil et al., 1998). The immunostimulant activity of bitter melon (Momordica charantia) leaves juice was known there was no documentary evidence.
Conclusion
Bitter melon (Momordica charantia) leaves juice has shown significant immunomodulatory effect in animals. Therefore clinical studies as a potential immunostimulat is further warranted. Therefore, we assume that clinical studies with bitter melon (Momordica charantia) leave juice in increased activity of immunoglobulin G (IgG) and immunoglobulin M (IgM) will result in positive outcome.
Acknowledgements
The authors are thankful to head, Faculty of Pharmacy, Hasanuddin University for providing the facilities and crew of department of immunological science, Medical Laboratory Makassar for helpful during this research.
References
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Bratawidjaja, K. 2004.Imunologi Dasar. Edisi VI. Fakultas Kedokteran Universitas Indonesia. Jakarta. 19, 78, 82.
Patil,J.S., B.G., Nagavi, M. Ramesh and G.S. Vijayakumar. 1988. A study on the immunostimulant activity of Centella asiatica in rats.Indian Drugs., 35:711-714 .
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Abascal K, Yarnell E. 2005.Using bitter melon to treat diabetes. Altern Complemen Ther 11(4):179-184
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