Ameliorative effect of phytoconstituents of Momordica charantia on body weight in Cerulein induced acute pancreatitis in Swiss albino mice

Introduction

Acute pancreatitis (AP) an inflammatory condition of the pancreas which appears at a rapid pace and cause severe inflammation of the pancreas. AP remains to be the 3 rd most common cause of immediate hospital admission under gastrointestinal diseases and is 6 th in position for the non-malignant cause of death, and it even acts as a major contribution factor for the development of chronic pancreatitis and pancreatic cancer (Peery et al., 2019) [1] . The pathogenesis of AP starts with the activation of intra-pancreatic acinar cell zymogen leading to conversion of inactive trypsinogen to active trypsin which cause dysregulated digestive enzyme secretion causing the accumulation of vacuoles and simultaneously triggering the inflammatory pathways which terminally end up in apoptosis and necrotic acinar cell death (Gukovskaya et al., 2019) [2] . The manifestation of the disease is a multistep process which starts with local limited inflammation and takes its turn towards systemic inflammation by release of diverse inflammatory mediators such as cytokines, reactive oxygen species (ROS), chemokines, leukocyte adhesion molecules, lipids and gaseous mediators (Granger and Remick, 2005) [3] . Cholecystokinin (CCK) analogue, cerulein is well known for induction of AP with high reproducibility. The supramaximal doses of cerulein result in dysregulated enzyme production, formation of vacuoles in the cytoplasm, edema formation and inflammatory cell infiltration into the pancreas (Lee et al., 2022) [4] . Momordica charantia Lam. (MC) is commonly called a bitter melon of the Cucurbitaceae family and is well known for its medicinal value and nutritional properties (Polito et al., 2016) [5] . The main chemical constituents include heteropolysaccharides, proteins and peptides (Schrot et al., 2015) [6] , terpenoids, saponins, flavonoids, phenolic compounds, essential oils, fatty acids, amino acids and sterols (Dandawate et al., 2016) [7] . Phenolic compounds present in the pericarp, stem and leaves serve as good antioxidants by directly acting on the ROS and even inducing the endogenous defence mechanisms (Yoshime et al., 2016) [8] . The glycosidic compounds from the fruit grouped as cucurbitine-type triterpenoids (cucurbitacins) are known to have anti-inflammatory and anti-diabetic properties (Rios et al., 2005) [9] .

Materials and Methods

The present experimental study was conducted on male Swiss albino mice of uniform age (about 6-8 weeks), which were procured from Jeeva Life Sciences, Hyderabad. Sterile mice feed in the form of pellets was purchased from M/s. Jeeva Life sciences, Hyderabad. All the drugs used in the study were purchased locally in Hyderabad. Fruit samples of Momordica charantia were thoroughly washed with tap water to remove any waste and dust particles. The excess water was removed by pressing with a paper towel gently and allowed to dry under shade for 3-4 weeks until constant weight is achieved. Once the samples were dried they were ground to semi powder form using a commercial grinder and were stored in an airtight polythene bags in the refrigerator at 4 °C for further analysis. Later, ethanolic extracts were prepared following the established protocol.

A total of 40 male Swiss albino mice of 6-8 weeks of age, weighing between 22-30 g. were obtained from Jeeva Life sciences, Hyderabad. The animals were kept in polypropylene cages and maintained with 12 hrs. dark/light cycle under hygienic conditions having ambient temperature (22-24 °C) for acclimatization (1 week) prior to the beginning of the experiment. Animals were placed on commercial standard pellet feed and provided water ad libitum throughout the experiment. The experimental protocol was approved by the Institutional Animal Ethics Committee, College of Veterinary Science, Rajendranagar, Hyderabad. All the groups were maintained as per the following treatment schedule for 7 days. The mice were randomly divided into 5 groups consisting of 6 mice in each group with the following schedule.

Experimental Design

Group-1

Normal control (NC)/Sham-normal saline for 7 days.

Group-2

Momordica charantia fruit extract control (MCFE)/ per se @ 400 mg/kg b.wt. through oral route.

Group-3 Cerulein control (CC)/Positive control/Disease control (DC) received 6 consecutive hourly intraperitoneal injections of cerulein @ 50 μg/kg b.wt. on 7 th day.

Group-4

Momordica charantia fruit extract (MCFE) low dose @ 200 mg/kg b.wt. + cerulean (Pre-treatment with Momordica charantia fruit extract (MCFE) low dose @ 200 mg/kg b.wt. + 6 consecutive hourly intraperitoneal injections of cerulein @ 50 μg/kg b.wt. on 7 th day).

Group-5

Momordica charantia fruit extract (MCFE) high dose @ 400 mg/kg b.wt. + cerulein Pre-treatment (Pre-treat with Momordica charantia fruit extract (MCFE) high dose @ 400 mg/kg + 6 consecutive b.wt. + 6 hourly intraperitoneal injections of cerulein @ 50 μg/kg b.wt. on 7 th day). Mice body weights were measured and recorded on the 7 th day.

Statistical analysis

The results of the experiment are presented as mean ± SEM (standard error of the mean) where multiple determinants of were considered and comparison among groups was analyzed using one-way analysis of variance (ANOVA) followed by Tukey's Multiple comparison test using GraphPad prism scientific software version 5.0. P-value < 0.05 was considered as significant.

Results and Discussion

Average body weight

The average body weight (g) in group 3 (27.83±1.38) was not significantly different from group 1 (28.00±1.69). Similar results were noticed in treatment groups 4 and 5 (27.00±0.55 & 28.80±1.16 respectively). Further, no significant difference was observed in the average body weights of mice among groups 1 and 2 (Table 1).

Discussion

The average body weights (g) of mice were not significantly altered among the groups.

This could be assigned to the reduced expression of inflammatory proteins and also could be because of activation of PPARα and PPARγ activators which are known for maintaining the blood lipids and also exhibit antiinflammatory activity through inhibition of cytokine production by activation of PPARγ (Ciou et al., 2014) [10] .

Conclusion

It is evident from the investigation that presence of terpenoids, saponins, flavonoids, phenolic compounds, essential oils, fatty acids, amino acids and sterols, cucurbitine-type triterpenoids (cucurbitacins) are known to have anti-inflammatory and anti-diabetic properties (Rios et al., 2005) (9) that might be attributed for maintainance of the body weight in Momordica charantia treated mice. Further, investigations are required to delineate the actual role of these phytoconstituents at molecular level.

References

1. Peery AF, Crockett SD, Murphy CC, Lund JL, Dellon ES, Williams JL, et al. Burden and cost of gastrointestinal, liver, and pancreatic diseases in the United States: update 2018. Gastroenterology. 2019;156(1):254-272.
2. Gukovskaya AS, Gorelick FS, Groblewski GE, Mareninov OA, Lugea A, Antonucci L, et al. Recent insights into the pathogenic mechanism of pancreatitis: role of acinar cell organelle disorders. Pancreas. 2019;48(4):459-462.
3. Granger J, Remick D. Acute pancreatitis: models, markers, and mediators. Shock. 2005;24:45-51.
4. Lee Y, Lim JW, Kim H. α lipoic acid inhibits cerulein/resistin induced expression of interleukin 6 by activating peroxisome proliferator activated receptor γ in pancreatic acinar cells. Molecular Medicine Reports. 2022;26(2):1-11.
5. Polito L, Bortolotti M, Maiello S, Battelli MG, Bolognesi A. Plants producing ribosome-inactivating proteins in traditional medicine. Molecules. 2016;21:1560-1563.
6. Schrot J, Weng A, Melzig MF. Ribosome-inactivating and related proteins. Toxins. 2015;7:1556-1615.
7. Dandawate PR, Subramaniam D, Padhye SB, Anant S. Bitter melon: a panacea for inflammation and cancer. Chinese Journal of Natural Medicines. 2016;14(2):81100.
8. Yoshime LT, de Melo ILP, Sattler JAG, de Carvalho EBT, Mancini-Filho J. Bitter gourd (Momordica charantia L.) seed oil as a naturally rich source of bioactive compounds for nutraceutical purposes. Nutrire. 2016;41:12-19.
9. Rios JL, Escandell JM, Recio MC. Studies in Natural Products Chemistry. Amsterdam: Elsevier; 2005. p. 429-437.
10. Ciou SY, Hsu CC, Kuo YH, Chao CY. Effect of wild bitter gourd treatment on inflammatory responses in BALB/c mice with sepsis. BioMedicine. 2014;4(3):1-7.