Laxative effect of green gedi leaves infuses (Abelmoschus manihot (l.) medik) on male white mice (Mus musculus)

Background: Gedi plant (Abelmoschus manihot (L.) Medik) of the Malvaceae family is a plant which leaf is used by peoples for the treatment of several illnesses such as constipation. Purpose: To investigate the laxative activity of green gedi leaves infuses (Abelmoschus manihot (L.) Medik) on male white


Introduction
Constipation is a chronic problem in many patients all over the world. In some groups of patients such as the elderly, constipation is a significant health-care problem, but in the majority of cases chronic constipation is an aggravating, but not life-threatening or debilitating, complaint that can be managed in primary care with cost-effective control of symptoms (WGO, 2010) Constipation is a syndrome that is defined by bowel symptoms (difficult or infrequent passage of stool, hardness of stool, or a feeling of incomplete evacuation) that may occur either in isolation or secondary to another underlying disorder (eg, Parkinson's disease). Although many physicians regard constipation as synonymous with reduced stool frequency, others also consider straining to defecate, hard stools, and the inability to defecate at will as constipation (Bharucha A. E., 2013) It is reported that the mean constipation rates in Europe and Oceania are 17.1% and 15.3% respectively. This prevalence seems to be majorly affected by female gender, age and socioeconomic and educational class (Peppas, et.al, 2008). It is mentioned that constipation makes 3% of patient visits to a general pediatrician and 15-25% visits to pediatric gastroenterologist (Kadim, M., 2015). For geriatric patients in Indonesia, the constipation prevalence is 3.8% for the elderly aged 60-69 years and 6.3% for the elderly over 70 year old (Kemenkes RI, 2013). Despite the high prevalence rate, constipation is usually temporary and harmless. Although in some severe cases it can indicates a serious underlying gastrointestinal lesions such as colorectal cancer (Longgo, D. et. al., 2015) Majority of the patients suffering from constipation will do their own treatment to improve the complaint, such as buying over the counter (OTC) laxatives.
Given the high prevalence of constipation, it is necessary to research about traditional medicine that can be used in constipation. Indonesia is the world's biodiversity mega-center. Among the 30.000 plant species living in the Indonesian archipelago, it is known that at least 9.600 species are efficacious as medicine, and approximately 300 species have been used as traditional medicinal materials by the traditional medicine industry (Depkes RI, 2007) Green gedi plant (Abelmoschus manihot (L.) Medik) is one of the many plant species that grows flourishly in Indonesia, especially in North Sulawesi. The local people of North Sulawesi commonly use its leaves as an ingredient for a traditional dish called "tinutuan" or Manado porridge. Green gedi leaves are not only used for cooking but also said to have efficacy in traditional medicine. As traditional medicine, it is said to be able treat many illness, starting from mild illness to severe illness, such as kidney disease, ulcers and high cholesterol.

Materials and method 1. Test material
The material tested is green gedi leaves (Abelmoschus manihot (L.) Medik) obtained from a yard of a house in Surabaya. Gedi leaves then dried and powdered at UPT Materia Medica Batu. Infuses are made by boiling the powder and aquades for 15 minutes and then filtered with flannel for the water to be taken.

Experimental Animal
White male mice (Mus musculus) balb/c with age around 3 months old, weight around 30 gram, and in a healthy physical condition, are obtained from Pharmacology Laboratory of Medical Faculty of Airlangga University Surabaya. Mice are acclimatized for a week.

Method
The mice that are used have normal feces characterization and fasted for 1 hour prior to the treatment. After the mice being weighed, categorized randomly into 5 groups with 6 mice in each group. Every group is given treatment as follow: Group I : administered with CMC Na 0.5 % only Group II : administered with infuses 130 mg/kgBW dose (half normal dose) Group III : administered with infuses 260 mg/kgBW dose (normal dose) Group IV : administered with infuses 520 mg/kgBW dose (twice normal dose) Group V : administered with bisacodyl All infuses are given orally. The observed variables are : frequency of defecation, weight and consistency of feces. Observation is done in 6 hours with 60 minutes interval. The frequency of defecation is observed within 6 hours with 60 minutes interval on how often the mice defecate. Consistency of feces is determined by the seepage diameter of the feces on the filter paper. Observation starts from after the administration of the treatments until the mice defecate, assessment of feces consistency is divided into: -0 = not defecating -1 = hard feces (H) with diameter on filter paper < 0.5 cm -2 = mushy feces (M) with diameter on filter paper 0.5 -1 cm -3 = liquid mushy (LM) with diameter on filter paper > 1 cm The weight of the feces is measured by collecting the feces on the filter paper that comes out within 6 hours with 60 minutes interval. According to table 1, administration of 520 mg/kgBW dose has the highest mean defecating frequency with a value of 7.17. There is a significant difference between the groups with administration of CMC Na 0.5% and 520 mg/kgBW dose as the p value is lesser than the alpha value (0.05). However, the ANOVA p value (0.132) in this table is greater than the alpha value (0.05), therefore there in no significant difference between the groups overall.  In LSD test however, there is significant difference in the frequency of feces between Na CMC 0.5% and 520 mg/kgBW dose with p. value of 0.038 and another significant difference in the weight of feces between 130 mg/kgBW and 260 mg/kgBW.dose with p. value of 0.034. Thus, pharmaceutically there is a difference because there is an increased of frequency of defecation for infuses compared to negative control. However, it is shown that in term of frequency of defecation, the result for green gedi leaves infuses 540 mg/kgBW dose is higher than 130, 260 mg/kgBW dose, negative control and even the positive comparative control (bisacodyl). This suggests that the higher the dosage of green gedi leaves infuses the higher the frequency of defecation in animal experiments.

Result
Based on the literature gathered, green gedi leaves contains polyphenol compounds, free steroids, saponins, flavonoids, essential oils, phenolic compound, steroid compound and gum mucilage. It has been widely known that mucilage is a type of soluble fiber with thick gluey characteristic and often used as colon cleanser because of its ability to absorb excess intestinal fluids, which softens stools and promotes regular bowel movement (Zliving, 2014). It is possible that these compounds contained in green gedi leaves are responsible for the laxative activity. According to Méité, S., et al. (2010), phytoconstituents like flavonoids, alkaloids, terpenoids, sterols, phenolic compounds, and tannins are responsible for laxative activities in plants.
Since there is no significant difference between the highest dose of infuses and bisacodyl, especially on the frequency of defecation, it might suggest that the laxative activity in green gedi infuses work similarly like bisacodyl by stimulating fluid secretion and promoting colon contraction to empty out the colon and excrete the feces thus giving a laxative effect on mice

Conclusion
From the results of research that has been done can be concluded as follows: 1. Green gedi leaves infuses (Abelmoschus manihot (L.) Medik) doesn't have a significant laxative effect on male white mice (Mus musculus) 2. The use of green gedi leaves as laxative by society cannot be proven empirically in the laboratory.