INTRODUCTION:

Plant and plant derivatives have long been a source of medication in human culture. According to world health organizations, 80% of the population of developing countries relies for their primary health treatment on conventional medicines. Medicinal plants may be a major source of undiscovered chemicals with significant therapeutic effects.¹ The growing value as potential preventive interventions of biologically active medicinal plants and their constituents has been a topic of intense research. In the future, safe and effective drugs are likely to be manufactured from medicinal plants to heal multiple degenerative diseases.

Constipation is a normal, sometimes persistent, gastrointestinal disorder in which the movement of small quantities of stiff, dry stool is fewer than three days a week or major changes in one's regular routine, followed by discomfort and feeling bloated, or abdominal fullness, impacting the quality of life. Constipation has been identified as one of the daily health issues which has recently been increased due to increased intake of processed foods or foods deficient in natural dietary fibres.^2,3,4,5

Even if not life-threatening, constipation and/or diarrhea have caused the quality of life to be significantly inconvenient. Functional intestinal disorder (FBD) has recently been identified as a middle or lower gastrointestinal tract-related symptom condition involving functional constipation, functional diarrhea, and irritable bowel syndrome (IBS), although it does not define the precise etiological basis of FBD. Distinguishable symptoms of FBD are diagnosed in lack of a physiological or biochemical explanation; the more frequent symptoms of IBS include stomach pain or irritation associated with intermittent constipation and diarrhea.^6,7 Constipation can be chronic or acute. Constipation is mostly acute and not harmful.⁸ Acute constipation may cause the intestine to close, and may require surgery as well.⁹ Chronic constipation is a complicated disorder of the elderly, marked by difficult stool movement.¹⁰ This condition has a strong correlation with patients 'quality of life¹¹ and health resource consumption.^12,13 In India's Ayurvedic medicine method the herb Capparis moonii wight is used for the treatment of various diseases. Capparis moonii, Wight belongs to the Capparidaceae family, commonly known as Large Caper, found in the Konkan regions, which in hot semiarid conditions grows vigorously.¹⁴ Its global distribution is limited to the Indian Subcontinent, i.e. Southern India, where Sri Lanka shows its endemism. Capparis moonii Wight, known in Marathi As Waghati, and as Rudanti in Sanskrit.¹⁵ Ayurveda defines Rudanti 's therapeutic applications, that nourish single body cell (Rasayani). That is effective under conditions of digestion and emaciation (Shoshghani). Symptoms of aging (Jara Vinashnam) are delayed by the use of rudanti and they are also useful for diseases which have devastating effects on all systems of the body (Rajyakshma Shasyate). Rudanti has been commonly used to get relief from asthma and cough and emerges as an incredibly useful bioprospecting herb because of the significant ethnomedicinal value of the capparis genus, Capparis moonii, Wight.¹⁶

Collection and authentication of plant:

The dried fruits of Capparis moonii wight were collected from Dawakhana, Pydhony, Bhuleshwar, Mumbai in July, 2019. Sample specimen voucher was submitted to Dr. Bindu Gopalkrishnan Asst. Professor at Mithibai College of Arts, Chauhan Institute of Science and Amruthben Jivanlal College of Commerce and Economics and authenticated and then the samples were sent to the Pharmacology Department, Oriental College of Pharmacy, Sanpada, Navi Mumbai- 400705.

Preparation of extract:

The dried fruits were grinded to make coarse powder. The powdered plant fruits of Capparis moonii wight was kept in the extraction chamber of soxhlet extractor with help of cotton plugs to filter the solvent returning into round bottom flask from crude powder. The soxhlet extractor was mounted on the ethanol (90%v/v) containing round bottom flask. The condenser was fixed at the other end of the soxhlet extractor at avoid solvent leakage due to vapor formation and vapor condensation. The solvent dissolves the powder's soluble ingredients, which changes colour. The siphon tube drained the chamber, with solvent flowing back into the round bottom flask. This process was repeated until the powder was exhausted which was detected in siphon tube by the presence of the clear liquid. In petri dishes, the contents of the round bottom flask were drained, and the solvent was allowed to evaporate. The solvent had been evaporated to obtain the extract's dried powder. These crude dry extracts have been placed in an appropriate container and kept in the refrigerator at 4°C until use. Alcoholic extract percentage yield was 7.88 per cent w/w. The ethanol extract of Capparis moonii wight was used for the entire study.

Qualitative phytochemical screening:

Preliminary chemical tests were carried out on ethanolic extract of Capparis moonii wight for determination of presence of phytoconstituents like alkaloids, carbohydrate, flavonoids, saponins, triterpenoid saponin, tannins and phenolic compounds.

Animals:

Either sex Swiss Albino Mice (25-30g) were used for the study. The animals were obtained from Bombay Veterinary College Parel, Mumbai 400012. The use of these animals and the study protocols were approved by CPCSEA recognized institutional ethics committee of Oriental College of Pharmacy under protocol no. OCP/IAEC/2018-2020/05 titled “evaluation of laxative activity of ethanolic extract of Capparis moonii wight fruits in swiss albino mice” under the thesis title “Evaluation of Pharmacological Activity of Medicinal Plant Extract”. Mice were kept at the well ventilated, air-conditioned animal house of Oriental College of Pharmacy, Sanpada, Navi Mumbai, In polypropylene cages, at 22 ± 2°C, with 12:12 hrs dark: light cycle. They were provided with commercial mice feed and water given ad libitum.

Acute oral toxicity¹⁷:

Acute oral toxicity study was performed as recommended in OECD Guideline 423.

Chemicals:

Dulcoflex 5mg (bisacodyl), Charcoal Meal (vegetable charcoal, 10% gum acacia)

Pharmacological activity:

A) Evaluation of Laxative activity:^18,19,20

Mice were fasted for 12 h before the experiment. fasted mice were placed individually in cages lined with clean filter paper. Mice were divided in five groups with the first group acting as the control and will receive saline that will act as the negative control. The second group will receive sodium picosulfate, this will serve as the positive control. The third, fourth and fifth groups will receive low, medium and high dose per os of the ethanolic extract of Capparis moonii. Immediately after dosing, the animal were examined for laxation for 5h with the withdrawal of food and water. The animals were kept for washing for 10days periods and then they were reuse for Charcoal meal gut transit test.

B) Charcoal meal gut transit test:^18,19,20

18 hr prior to the experimentation food were withdrawn but not water. The drugs were administered 60 min before administration of charcoal meal. Animals were sacrificed after various time intervals. Entire intestine were remove immediately and were immerse in 5% formalin. Then distance between pylorus region and front of charcoal meal were measured and evaluated by student t-test.

Statistical analysis:

All the data expressed are Mean ± Standard Error of Mean (S.E.M.) with 95% confidence intervals. One way analysis of variance (ANOVA) followed by Dennett’s test was used to assess the laxative activity and Charcoal meal transit test. All the graphs, calculations and statistical analysis were performed using prism 8.4.1(676) Software by Graph Pad. Where, * represents significant at p≤0.05, ** represents highly significant at p≤0.01, as compared with intoxicant control; ***represents highly significant at p≤0.001 on comparison with Normal control and**** represents highly significant at p≤0.0001 on comparison with Normal control.

RESULTS:

Qualitative phytochemical screening:

Qualitative phytochemical screening of the ethanolic extract of C. moonii revealed presence Alkaloid, flavonoids, saponins, glycosides, tannins, phenolic compounds, terpenoids, Carbohydrates and proteins were absent. The presence of flavonoids and saponins, glycosides in the extract, significantly correspond to the laxative activity of ethanolic extract. of Capparis moonii Wight.

Table No. 1: Result of Quantitative Phytochemical Analysis of Powdered Fruits of Capparis moonii wight

Sr. No.	Phytoconstituents	Ethanolic extract of the leaves Trema orientalis L.
1	Alkaloids	+
2	Flavonoids	+
3	anthraquinone Glycosides	+
4	Saponins	+
5	Steroids	+
6	Terpenoids	-
7	Tannins and Phenolic compounds	+
8	Carbohydrates	+
9	protein	-

Present (+)/Absent (-)

Acute oral toxicity studies:

As suggested in OECD Guideline 423, acute oral toxicity study was conducted and the results revealed that no toxic signs were observed in clinical parameters during an acute toxicity test of up to 2000mg / kg. It thus indicates that the median lethal dose (LD50) of alcoholic extract of dried fruit Capparis moonii wight is equal to 2000 mg/kg.

1) Effect on extracts on laxative activity:

The ethanolic extract Capparis moonii was studied for its laxative activity. The laxative activity was assessed by measuring the wet faeces in all test groups. The statistical significance of difference between the data of test groups was calculated and values are present in table 2. In this study, the different doses of the extract showed dose dependant increase in fecal output of rats when compared to the control group. Extract showed increase in fecal output of 1 mice when compared to control group. The effect of extract at dose of 50,100,200mg/kg increased significantly fecal output of mice as compared to control group. The effect of Capparis moonii at dose of 200mg/kg was comparable to that of standard drug dulcoflex (5 mg/kg, p.o.).

Table No. 2: Faeces Output in Mice

Sr No.	Groups	Drug Treatment	Faeces out put
1.	Control	Distilled Water (2ml/kg)	1.98±0.29
2.	Standard	Dulcoflex (5mg/kg)	6.13±0.57^****
3.	Low Dose	CMEE(50mg/kg)	3.70±0.33^*
4.	Medium Dose	CMEE (100mg/kg)	4.2±0.2^**
5.	High Dose	CMEE(200mg/kg)	4.9±0.65^***

Values are in Mean ± SEM(n=6); *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 Vs Control

Bar diagram:

Fig. 1: Laxative Activity in Mice

Bar diagram showing the Dose dependent effect of EECM on the faeces output in mice. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001 show a comparison of group 2,3,4,5vs group1(One way ANOVA followed by Dunnett’s test). Each bar shown represents mean ± SEM of 6 animals per group.

2) Effect on extracts on gastro intestinal motility:

The passage of charcoal meal through the gastro intestinal tract in mice is used as parameter for intestinal motility and to study the effect of laxatives. The effect of ethanolic extract of Capparis moonii on gastrointestinal motility was tested on mice as per the protocol mentioned in materials and methods. The distance travelled by charcoal meal from the pylorus to the caecum was measured, and expressed as the percent of the total length of the distance. The ethanolic extract of Capparis moonii increased propulsion of the charcoal meal through the gastrointestinal tract in a concentration dependant manner The ethanolic extract of Capparis moonii at dose of 200mg/kg significantly increased in the propulsion of charcoal meal as compared to control group. The result of motility test is reported in table 3.

Bar diagram:

Fig. 2: Percentage Length of Small Intestine Travelled by Charcoal Meal

Bar diagram showing the Dose dependent effect of EECM on the travel of charcoal meal through small intestine of mice. *p<0.05 ,**p<0.01,***p<0.001,****p<0.0001show a comparison of group 2,3,4,5vs group1. Each bar shown represents mean±SEM of 6 animals per group.

DISCUSSION:

The present research was carried out to evaluate the laxative effects of ethanoic extract of Capparis moonii wight fruit. The acute oral toxicity analysis showed that ethanoic extract of Capparis moonii wight fruit was safe for administration in rats/mice at the highest dose (2000mg/kg) tested. The Capparis moonii wight’s axative activity was studied in mice. The result showed that an oral administration of Capparis moonii wight fruit extract produced a significant and dose-dependent increase in stool count in mice, as well as a dose dependent effect through distance traveled by charcoal meal. These effects were comparable with dulcoflex (bisacodyl) (standard drug) at dose of 5mg/kg. Bisacodyl is widely used laxative stimulant to manage constipation. It is presumed that the therapeutic effect of bisacodyl is due to its motility and secretory properties.²¹ The current preliminary phytochemical analysis also revealed in support of earlier reports that Capparis moonii wight shows the presence of compounds of glycoside, flavonoids, and saponins. Anthraquinone glycoside are reported to be present in other species of capparidaceae family. 22 Derivatives of anthraquinone show a wide range of pharmacological activities including laxative.^23,24,25 Glycosides (Anthraquinone) exhibit laxative effect through the following mechanism: Changes in absorption, excretion, secretion leading to diarrhea due to accumulation of fluid and Changes the colonic motility contributing to rapid large intestine transit.^26,27 Flavonoids have been used in the treatment of gastrointestinal disorders along with medicinal properties such as laxative and effective in constipation.^28,29,30 Several reports on the effects of saponin- containing extracts and compounds have shown that steroidal saponin may contribute to a reduction in constipation symptoms.³¹ Prior to this, the presence of phytoconstituents such as terpenoids, sterols, flavonoids, phenolic compounds was observed to be responsible for laxative activities in plants.³² Charcoal meal propulsion is probably due to the increasing peristaltic motion in the gastrointestinal tract of mice resulting from C. moonii 's stimulation of cholinergic receptors. Neural and myogenic mechanisms control the intestinal movement.³³ An rise in the contractile activity of the smooth layers in general is responsible for increasing the propulsion of intestines. These movement behaviors are regulated by many Mediators and Neurotransmitters. Acetylcholine is the principal stimulating neurotransmitter of the enteric nervous system.³⁴ Thus the plant extract can explain the usefulness of Capparis moonii in constipation.

CONCLUSION:

From the result it can be concluded that oral administration of Capparis moonii wight ethanolic extract indicates significant laxative activity in mice by faeces output and percentage length of small intestine traveled by charcoal meal. This study has shown that Capparis moonii wight fruit ethanolic extract has laxative effects in addition to the various physiological effects earlier reported by other authors.

ACKNOWLEDGMENT:

We are thankful for their guidance and help to our Principal Dr.(Mrs.) Sudha Rathod, Dr. (Mrs.) Vanita G. Kanase, as well as to the Department of Pharmacology, Oriental College of Pharmacy, Navi Mumbai.

AUTHORS’ CONTRIBUTIONS:

Dr. (Mrs.) Vanita Kanase guided with designing the study, making of protocol and managed the work done. Shalaka Kadam performed the literature searches, performed the acute toxicity, models, phytochemical screening and completed the manuscript writing.

CONFLICTS OF INTEREST:

We announce we do not have conflicting interests.

AUTHORS’ FUNDING:

We thank Oriental College of Pharmacy for funding the project.

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Received on 18.05.2020 Modified on 02.08.2020

Research J. Pharm. and Tech. 2021; 14(7):3528-3532.

DOI: 10.52711/0974-360X.2021.00611

Groups	Drug Treatment	Charcoal Dose	% length of charcoal moved		P Value
Groups	Drug Treatment	Charcoal Dose	60 min	120 min	P Value
1. Control	Distilled Water (2ml/Kg)	0.2ml	65.34±0.05	68.27±0.49
2. Standard	Dulcolax (5mg/Kg)	0.2ml	86.23±0.72	95.46±0.40	<0.0001^****
3. Low Dose	E.Cs (50mg/Kg)	0.2ml	80.465±0.14	85.54±0.26	<0.0001^****
4. Medium Dose	E.Cs ( 50mg/Kg)	0.2ml	83.34±0.26	90.5±0.18	<0.0001^****
5. High Dose	E.Cs ( 50mg/Kg)	0.2ml	86.20±1.5	94.86±0.57	<0.0001^****