Open Veterinary Journal, (2026), Vol. 16(4): 2313-2319

Research Article

10.5455/OVJ.2026.v16.i4.31

---

The laxative efficacy of a mixed extract of Cassia alata with Plantago major in rats

Amaq Fadholly^1*, Siti Sadiah¹, Rudi Heryanto² and Maharani Kartika Ramadhan³

¹Division of Pharmacology and Toxicology, School of Veterinary Medicine and Biomedical Sciences, IPB University, Bogor, Indonesia

²Department of Chemistry, Faculty of Mathematics and Natural Sciences, IPB University, Bogor, Indonesia

³Doctoral Program in Animal Biosciences, Department of Biology, Faculty of Mathematics and Natural Sciences, IPB University, Bogor, Indonesia

*Corresponding Author: Amaq Fadholly. Division of Pharmacology and Toxicology, School of Veterinary Medicine and Biomedical Sciences, IPB University, Bogor, Indonesia. Email: amaqfadholly [at] apps.ipb.ac.id

Submitted: 03/10/2025 Revised: 21/02/2026 Accepted: 03/03/2026 Published: 30/04/2026

---

ABSTRACT

Background: The treatment of constipation with contemporary medications is costly and has a significant risk of adverse effects.

Aim: This study evaluated the combined efficacy of the ethanolic leaf extract of Cassia alata with Plantago major in alleviating loperamide-induced constipation in rats.

Methods: Rats had constipation induced by an oral administration of loperamide (3 mg/kg BW) for 6 days, followed by three control groups assessment (normal, negative, positive which treated bisacodyl), and three treatment groups of the leaf extract of C. alata with P. major (50, 75, and 100 mg/kg BW) for 6 days. The alterations in quantity, mass, and hydration of feces, motility, and gastrointestinal transit ratio were calculated.

Results: All groups of the treatment demonstrated that oral administration of 50–100 mg/kg of the leaf extract combination demonstrated significant laxative activity, as reflected by improved gastrointestinal transit.

Conclusion: The leaf extract combination of C. alata with P. major, with dosages of 50–100 mg/kg, has potential laxative properties. This combination at the minimum dosage of 50 mg/kg is more favorable, potentially due to a synergistic interaction between the two extracts. Further studies involving additional dose combinations are recommended.

Keywords: Cassia alata, Herbs, Laxative, Plantago major.

---

Introduction

Constipation is a prevalent and often chronic functional gastrointestinal disorder affecting 3%–15% of the general population (Al-Nou'mani et al., 2023). Constipation is defined by symptoms including difficult or infrequent bowel movements, stool hardness, and a feeling of incomplete evacuation associated with discomfort (Jani and Marsicano, 2018). Functional constipation consists of two subtypes: slow-transit constipation and dyssynergic defecation. Some people with irritable bowel syndrome may exhibit traits of both types of constipation (Bellini et al., 2013). Stool softeners, osmotic agents, bulking agents, and stimulant laxatives are frequently utilized to alleviate constipation. However, laxatives may precipitate cardiac problems and arterial constriction due to electrolyte depletion, particularly potassium; excessive use can lead to unstable cardiac function. It may disrupt their daily routines and overall well-being (Ishiyama et al., 2019).

The therapy of constipation with currently available medications is associated with negative effects. 20%–30% of persons employ numerous laxatives on a weekly basis (Bassotti et al., 2023). Medications containing sennoside or magnesium oxide exhibit strong laxative effects and are mostly indicated for constipation-related disorders. Additionally, they may induce negative effects, such as severe diarrhea, and extended use can result in melanosis coli, a risk factor for colon neoplasia (Lim et al., 2019). The results indicate that certain individuals may not exhibit contentment with their medicine, while others fail to attain complete healing. The advancement of safer and more effective medicines, marked by sufficient efficacy, less side effects, and lower costs, is of substantial importance (Kessoku et al., 2023).

Local society prefers traditional medicine derived from natural sources for treating various ailments, including constipation, due to its cost-effectiveness, accessibility, affordability, and the plethora of options it offers compared to synthetic pharmaceuticals (El Kutry and Sopeah, 2020; Alshehri et al., 2022; Wresdiyati et al., 2023; Andriyanto et al., 2025). The conventional use of Cassia alata and Plantago major as laxatives is acknowledged in the traditional management of many diseases (Aslam, 2021; Zhang et al., 2021). The leaves of C. alata have been recorded to possess many pharmacological activities, such as anti-inflammatory, analgesic, antiviral, antibacterial, and antiplatelet-aggregating actions (Colin et al., 2024). The leaves of C. alata contain several compounds, including flavonoids (kaempferol, quercetin, emodin), terpenoids, fatty acids, alkaloids, saponins, steroids, and anthraquinones (alatinone, chrysaphanol) (Saptarini et al., 2024). Anthraquinones constitute a category of stimulant laxatives that exert a direct influence on enterocytes, the enteric nervous system, and smooth muscle within the gastrointestinal tract. The prior research indicated that anthraquinone-containing products are predominantly utilized as laxatives (Zhao and Zheng, 2023). Further research demonstrates that C. alata leaf extract is safe for administration in rats at doses up to 2 g/kg body weight (Fadholly and Ramadhan, 2025). The leaf of P. major has several pharmacological activities, including antioxidant, anti-inflammatory, immune-enhancing, antibacterial, and wound-healing effects, due to compounds such as flavonoids and iridoid glycosides, notably aucubin and acteoside (Adom et al., 2017). The polyholosidic fraction extracted from the leaves of some Plantago species (Plantago major, media, and lanceolata) accounts for its gastroprotective properties and has a laxative effect at elevated doses (Najafian et al., 2018). The potential use of C. alata and P. major leaves together as a laxative will augment their synergistic effect. This research was driven by the ongoing endeavor to develop safe, effective, and economical plant-based laxatives. This study assessed the synergistic laxative effects of a leaf extract combination of C. alata and P. major in rats, juxtaposing the findings with those of bisacodyl, a prevalent laxative in Indonesia.

---

Material and Methods

Study period and location

This research was performed from April to August 2025 at the Laboratory Animal Management Unit, School of Veterinary Medicine and Biomedical Sciences, IPB University.

Preparation of leaf extract of C. alata and P. major

The leaf of C. alata and P. major were collected and authenticated at the Tropical Biopharmaca Research Centre, IPB University, Indonesia. Four hundred grams of C. alata and P. major leaf simplicias were combined (1:1). The simplicia were prepared via maceration using 70% ethanol, with a simplicia to solvent ratio of 1:10. This was reconstituted in distilled water to achieve the selected experimental dosage levels of 50, 75, 100 mg/kg BW for the experiment (dose selection based on previous research that C. alata leaf extract has an effective dose in 100 mg/kg BW) (Isdoni et al., 2025). Subsequently, the leaf extract of C. alata with P. major leaf extract were tested for phytochemical screening.

Experimental animals

Fourty healthy male Sprague Dawley rats, aged 6 weeks, were acquired from the Laboratory Animal Management Unit at the SSchool of Veterinary Medicine and Biomedical Sciences, IPB University. Rats were maintained in controlled environmental settings, with a temperature of 22℃–25°C, relative humidity of 50%–60%, and a 12:12 hour light–dark cycle. Animals were housed in ordinary polypropylene enclosures with wood-chip bedding. The rats were administered a commercially standard laboratory diet with a balanced nutritional profile (proteins, carbs, fats, vitamins, and minerals), and water was available ad libitum. All rats were categorized into six groups (n=5) based on body weight (mean: 225.50 ± 10.50 g, range: 180.22–250.12 g) and fecal water content (mean: 28.13% ± 3.42%, range: 23.33%–38.11%), measured 1 day prior to the administration of the initial dosage of the test substance. Group one (P0) serves as the normal control (not subjected to loperamide and untreated), group two (P1) functions as the negative control (subjected to loperamide, treated with distilled water), group three (P2) acts as the positive control (subjected to loperamide, treated with bisacodyl), while groups four, five, and six are treatment groups that are subjected to loperamide and treated with leaf extract of C. alata combined with P. major at dosages of 50 mg/kg BW (P3), 75 mg/kg BW (P4), and 100 mg/kg BW (P5). The medication was administered orally once daily for 6 days, 1 hour after the administration of loperamide.

Induction of constipation

Loperamide was administered orally at a dosage of 3 mg/kg for 6 days to induce constipation, 1 hour before the administration of each test substance (Lim et al., 2019).

Body weight measurement

The daily body weights of individual rats were recorded from 1 day prior to the administration of the test substances until the sixth day of administration. All rats underwent overnight fasting (water was supplied; 18 hours) before the initial treatment and at termination to minimize fluctuations in weight related to nutrition for the measurement of intestinal transport (Lim et al., 2019).

Fecal measurement

The fecal pellets were collected throughout a 24-hour period, 1 day prior to the initial treatment of the test substance, and immediately following the fourth administration for an additional 24 hours. The aggregate quantity, moisture content, and wet mass of the fecal pellets were computed. The gathered fecal pellets were desiccated at 60°C in a conventional drying oven for 24 hours to determine the fecal dry weights (Isdoni et al., 2025).

Intestinal transit ratio measurement

The rats underwent an 18-hour fasting period before the experiment. Ten minutes following the final injection of the test substance (sixth day of treatment), the animals in each group were administered 1 ml of a norit. Thirty minutes post-administration of the norit, a ketamine–xylazine combination was used for euthanasia (Isdoni et al., 2025).

Data analysis

Data were expressed as means ± SD from six replicates and evaluated via one-way analysis of variance, succeeded by the Duncan multiple range test to ascertain significant differences across all parameters. Values were considered statistically significant at p < 0.05 (Ishiyama et al., 2019).

Ethical approval

All treatment procedures were conducted under the auspices of the Animal Ethics Committee, School of Veterinary Medicine and Biomedical Sciences (SVMBS), IPB University, Indonesia (permission reference number: 342/KEH/SKE/VII/2025). All measures were implemented to alleviate animal distress during this experiment.

---

Results

Phytochemical screening of the leaf extract of C. alata with P. major

The phytochemical examination of the leaf extract of C. alata in conjunction with P. major indicates the presence of flavonoids, tannins, saponins, and terpenoids. The results of the phytochemical analysis are presented in Table 1.

Table 1. Phytochemical screening results of the leaf extract of C. alata with P. major.

Body weight measurement

The body weights were observed into three phases. In the first phase, 1 day before treatment, the weight of all groups averaged 225 g. The second phase, 1 day after treatment, showed that the weight of P1, P2, P3, P4, and P5 groups had a mean decrease of 20 g. The third phase, 6 days after treatment, showed that P1, P3, P4, and P5 showed a weight increase of 25–30 g, while only P2 exhibited a significant difference in body weight gain. P0 has a stable weight gain. The results of body weight gain of all groups are shown in Table 2 and Figure 1.

Fig. 1. Body weight gains in the constipation rats induced loperamide treated by the leaves extract combination of C. alata with P. major.

Table 2. Body weight gains in the constipation rats induced loperamide treated by the leaf extract of C. alata with P. major.

Fecal measurement

All fecal parameters (fecal wet and dry weight, fecal number, and water content) were evaluated. The results of fecal parameters along 24 hours before treatment were similar in all groups. However, P1 groups have significant decreases in fecal number and water content, which were detected following the fourth treatment day (after 24 hours) compared to P2, P3, P4, and P5 groups. The P0 group has stable fecal parameters. The results of fecal parameters of all groups are shown in Table 3.

Table 3. Fecal parameter in the constipation rats induced loperamide treated by the leaf extract of C. alata with P. major.

Intestinal transit ratio measurement

The results of a subsequent assessment of laxative efficacy using the intestinal transit method revealed that the leaf extract of C. alata and P. major in all treatment groups demonstrated a statistically significant difference compared to the negative control group, as it yielded results comparable to those of Bisacodyl in terms of marker-stained intestinal length (Table 4).

Table 4. Gastrointestinal transit ratio in the constipation rats induced loperamide treated by the leaf extract of C. alata with P. major.

---

Discussion

Constipation may result from various factors, including dietary practices, pharmacological agents like morphine, and psychological stress. The medicinal plants of C. alata and P. major are traditionally used in the treatment of constipation (Lizuka and Hamamoto, 2015). These plants expected synergistic effects compared to a single plant. This study is warranted due to the lack of prior investigations evaluating the combined effects of these plants, specifically about their reported therapeutic efficacy in the management of constipation. The application of loperamide as a constipating drug is well established. Loperamide is an opioid agonist used as an antidiarrheal medication that suppresses intestinal water output and colonic peristalsis (Stevick et al., 2023). Therefore, we used loperamide to induce constipation with oral administration (3 mg/kg) once daily for 6 consecutive days, 1 hour before the administration of the test drug, consistent with our prior research. The dosages of the leaf extract of these plants (50–100 mg/kg) were determined based on our previous study that the C. alata leaf extract have the optimum effect as laxative in 100 mg/kg on rats, therefore by combining it with P. major leaf, it is expected that the optimum dose obtained will be lower than with a single plant (Fadholly et al., 2025; Isdoni et al., 2025).

Plant extracts can be employed in the production of herbal medicine to isolate their phytonutrients (Abubakar and Haque, 2020; Arsyad et al., 2023; Aurellia et al., 2025). Phytochemical analysis for screening phytonutrients is a critical step in investigating the potential of medicinal plant resources across many categories of secondary metabolites, including triterpenoids, alkaloids, flavonoids, and steroids (Dubale et al., 2023). The phytochemical examination of the leaf extract of C. alata and P. major indicates the presence of flavonoids, tannins, saponins, and terpenoids. The phytochemical nature of this extract enables its function as a laxative. Flavonoids are thought to exert a laxative effect by significantly increasing intestinal peristalsis to facilitate digestion (Panche et al., 2016; Julián-Flores et al., 2025). Tannins enhance the intestinal milieu and regulate motility, whilst saponins function akin to stool-softening surfactants by diminishing surface tension and augmenting water infiltration into fecal matter (Kim et al., 2021). Terpenoids can activate intestinal smooth muscle and excite nerve plexuses, therefore augmenting motility (Zhao et al., 2021). The simultaneous presence of flavonoids, saponins, tannins, and terpenoids indicates a multifaceted mechanism whereby increased peristalsis, enhanced stool hydration, and stimulation of intestinal secretions yield a more potent laxative effect. Flavonoids and saponins are two secondary metabolites that markedly influence intestinal motility, emphasizing mechanisms that promote bowel movement. In this case, flavonoids include anthraquinone in C. alata, functioning as an adjunct laxative by indirectly enhancing intestinal motility. This shell increases stool volume and flexibility. Saponins operate as stimulant laxatives by directly increasing intestinal motility and secretion through the stimulation of sensory nerve endings in the intestinal wall (Kim et al., 2019). The phytochemicals, through complementary mechanisms that activate motility and regulate water and electrolyte balance, presumably jointly enhance fecal parameters and gastrointestinal transit as seen in our study. Table 2 indicates that the assessment of body weight revealed no significant alterations in body weight or weight increase in relation to the negative control (loperamide induction only) when compared to the normal control. The results are comparable across three distinct dosages of leaf extract from C. alata combined with P. major (50, 75, and 100 mg/kg) when compared to the negative control. The extract did not cause significant diarrhea as a side effect. Furthermore, it exerted no influence on body weight or weight increase. All groups involved in this investigation, including both control and treatment groups, demonstrated body weight gains within the anticipated range for age-matched rats (Kola-Mustapha et al., 2019).

Constipation typically leads to significant decreases in fecal output. The prolonged presence of fecal pellets in the extensive intestinal lumen may result in excessive water absorption, thereby reducing the water content of the excreted pellets. Consequently, fecal attributes such as the water content in feces and the quantity of pellets expelled serve as valuable markers for evaluating the efficacy of various laxatives (Włodarczyk et al., 2021). Loperamide has been shown to significantly reduce fecal water content and fecal pellet count, indicative of constipation (Hao et al., 2023). In comparison to negative control groups, rats administered with the leaf extract of C. alata combined with P. major (50–100 mg/kg) exhibited increased fecal water content and a greater number of ejected fecal pellets, signifying significant laxative efficacy on spastic constipation.

The gastrointestinal transit ratio, an indicator of intestinal motility, has been shown to decrease with loperamide, correlating with symptoms of spastic constipation (Papale et al., 2024; Parkar et al., 2024). The negative control groups in this study that received loperamide also exhibited the same symptoms. In comparison to the positive control group (bisacodyl), rats administered the leaf extract exhibited considerable enhancements in intestinal motility, as shown by the norite transit ratio. Thus, the observation that rats administered with the leaf extract of C. alata combined with P. major (50–100 mg/kg) demonstrated significant and dose-dependent enhancements in the gastrointestinal transit ratio relative to the negative control group indicates that this leaf extract possesses potential laxative properties against loperamide-induced constipation. It is advisable to do more investigations with alternative dose combinations.

---

Conclusion

The current study found that oral administration of 50–100 mg/kg of the leaf extract of C. alata with P. major demonstrated potential laxative effects, mediated by an increase in gastrointestinal motility in rats suffering from loperamide-induced constipation. The combined extract of C. alata and P. major (50–100 mg/kg) showed significant laxative activity in rats, suggesting a possible synergistic effect. Additional toxicological and mechanistic studies are recommended to confirm safety and efficacy.

---

Acknowledgments

The author expresses gratitude to the Directorate of Research and Innovation, IPB University, for the funding from the IPB Community Fund 2025 under Penelitian Dosen Muda 2025 scheme (Reference Number: 13465/IT3.D10/PT.01.03/P/B/2025).

Conflict of interest

The authors declare no conflicts of interest related to any financial, personal, or other relationships with individuals or organizations pertinent to the topic discussed in the work.

Author’s contributions

AF: Conception, Data curation, Formal analysis, Writing original draft, Supervision. SS: Data acquisition, Data Curation, Review Manuscript. RH: Data acquisition, Validation MKR: Supervision, Conception, Validation.

Data availability

All data that support the findings of this study are included within the manuscript.

---

References

Abubakar, A.R. and Haque, M. 2020. Preparation of medicinal plants: basic extraction and fractionation procedures for experimental purposes. J. Pharm. Bioallied. Sci. 12(1), 1–10.

Adom, M.B., Taher, M., Mutalabisin, M.F., Amri, M.S., Kudos, M.B.A., Sulaiman, M.W.A.W., Sengupta, P. and Susanti, D. 2017. Chemical constituents and medical benefits of Plantago major. Biomed. Pharmacother. 96, 348–360.

Al-Nou'mani, J., Al-Alawi, A.M., Al-Maqbali, J.S., Al-Abri, N. and Al-Sabbri, M. 2023. Prevalence, recognition, and risk factors of constipation among medically hospitalized patients: a cohort prospective study. Medicina (Kaunas) 59(7), 1347.

Alshehri , M.M., Quispe, C., Herrera-Bravo, J., Sharifi-Rad, J., Tutuncu, S., Aydar, E.F., Topkaya, C., Mertdinc, Z., Ozcelik, B., Aital, M., Kumar, N.V.A., Lapava, N., Rajkovic, J., Ertani, A., Nicola, S., Semwal, P., Painuli, S., González-Contreras, C., Martorell, M., Butnariu, M., Bagiu, I.C., Bagiu, R.V., Barbhai, M.D., Kumar, M., Daştan, S.D., Calina, D. and Cho, W.C. 2022. A review of recent studies on the antioxidant and anti-infectious properties of senna plants. Oxid. Med. Cell. Longev. 2022, 6025900.

Andriyanto, A., Aurellia, S., Putra, H.Y., Fadholly, A., Mustika, A.A., Sutardi, L.N., Pristihadi, D.N., Subangkit, M. and Ariyaliz, T. 2025. Evaluation of acute toxicity, sexual and stamina performance in mice administered with ashwagandha (Withania somnifera) suspension. J. Med. Pharm. Chem. Res. 7(11), 2397–2410.

Arsyad, R., Amin, A. and Waris, R. 2023. Technique of manufacture and values of simplicia and ethanol extract of bagore (Caesalpinia crista L.) seeds original polewali mandar. Makasar. Nat. Prod. J. (MNPJ). 3(14), 138–147.

Aslam, M.S. 2021. Application of genus Cassia in the treatment of constipation: a systematic review. F1000Research 8, 256.

Aurellia, A., Putra, H.Y., Fadholly, A., Mustika, A.A., Pristihadi, D.N., Subangkit, M. and Aryaliz, T. 2025. Evaluation of acute toxicity in mice administered with ashwagandha (Withania somnifera) suspension. J. Med. Pharm. Chem. Res. 7(11), 2397–2410.

Bassotti, G., Villanacci, V. and Corsetti, M. 2023. Exploring pharmacological treatments for chronic idiopathic constipation in adults: a look back to the future. J. Clin. Med. 2(4), 1702.

Bellini, M. 2015. Irritable bowel syndrome and chronic constipation: fact and fiction. World. J. Gastroenterol. 21(40), 11362–11170.

Bellini, M., Usai-Satta, P., Bove, A., Bocchini, R., Galeazzi, F., Battaglia, E., Alduini, P., Buscarini, E., Bassotti, G. and ChroCoDiTE Study Group, AIGO. 2017. Chronic constipation diagnosis and treatment evaluation: the "CHRO.CO.DI.T.E." study. BMC Gastroenterol. 17(1), 11.

Colin, M.N., Claudiana, N.S.E., Kaffah, A.U., Hasanah, A.N., Megantara, S. 2024. Review on Cassia alata bioactive compounds: in silico, in vitro, and in vivo studies. Drug Des. Devel. Ther. 18, 4427–4447.

Dubale, S., Kebebe, D., Zeynudin, A., Abdissa, N. and Suleman, S. 2023. Phytochemical screening and antimicrobial activity evaluation of selected medicinal plants in Ethiopia. J. Exp. Pharmacol. 15, 51–62.

El Kutry, M.S. and Sopeah, H.R.A. 2020. Impact of intake of Sidr (Zizyphus spina-christi L.) extract on enterobiasis vermicularis infection in children. Biome. Pharmacol. J. 13(1), 359–366.

Fadholly, A. and Ramadhan, M.K. 2025. Acute toxicity test of Cassia alata leaves extract on rats (Rattus norvegicus). Acta. Vet. Indones. 13(2), 100–105.

Fadholly, A., Sa’Diah, S., Pristihadi, D.N., Harlina, E., Ramadhan, M.K., Budiharjo, Y.A.K., Adinugroho, C.R., Rahmah, A.A. and Azizah, N.F. 2025. Unveiling the safety of Cassia alata leaf extract: a subchronic oral toxicity in rats. Adv. Anim. Vet. Sci. 13(4), 817–823.

Hao, M., Song, J., Zhai, X., Cheng, N., Xu, C., Gui, S. and Chen, J. 2023. Improvement of loperamide-hydrochloride-induced intestinal motility disturbance by Platycodon grandiflorum polysaccharides through effects on gut microbes and colonic serotonin. Front. Cell. Infect. Microbiol. 13, 1105272.

Isdoni, I., Fadholly, A., Sa'Diah, S., Mustika, A.A., Angelina, T. and Nanginda, A.M. 2025. Laxative potential of Cassia alata leaves extract: _{in vivo} study. Res. J. Pharm. Tech. 18(7), 2967–1970.

Ishiyama, Y., Hoshide, S., Mizuno, H. and Kario, K. 2019. Constipation-induced pressor effects as triggers for cardiovascular events. J. Clin. Hypertens. (Greenwich). 21(3), 421–425.

Jani, B. and Marsicano, E. 2018. Constipation: evaluation and management. Mo. Med. 115(3), 236–240.

Julián-Flores, A., Aguilar-Zárate, P., Michel, M.R., Sepúlveda-Torre, L., Torres-León, C., Aguilar, C.N. and Chávez-González, M.L. 2025. Exploring the therapeutic potential of medicinal plants in the context of gastrointestinal health: a review. Plants 14(5), 642.

Kessoku, T., Misawa, N., Ohkubo, H. and Nakajima, A. 2023. Current treatment practices for adult patients with constipation in Japan. Digestion 105(1), 1–9.

Kim, J.E., Park, J.W., Kang, M.J., Choi, H.J., Bae, S.J., Choi, Y.S., Lee, Y.J., Lee, H.S., Hong, J.T. and Hwang, D.Y. 2019. Anti-inflammatory response and muscarinic cholinergic regulation during the laxative effect of Asparagus cochinchinensis in loperamide-induced constipation of SD rats. Int J Mol Sci. 20(4), 946.

Kim, J.E., Choi, Y.J., Lee, S.J., Gong, J.E., Lee, Y.J., Sung, J.E., Jung, Y.S., Lee, H.S., Hong, J.T. and Hwang, D.Y. 2021. Antioxidant activity and laxative effects of tannin-enriched extract of Ecklonia cava in loperamide-induced constipation of SD rats. PLoS One 16(2), 246363.

Kola-Mustapha, A.T., Ghazali, Y.O., Ayotunde, H.T., Atunwa, S.A. and Usman, S.O. 2019. Evaluation of the antidiarrheal activity of the leaf extract of Parquetina nigrescens and formulation into oral suspensions. J. Exp. Pharmacol. 11, 65–72.

Lim, J.M., Kim, Y.D., Song, C.H., Park, S.J., Park, D.C., Cho, H.R., Jung, G.W., Bashir, K.M.I., Ku, S.K. and Choi, J.S. 2019. Laxative effects of triple fermented barley extracts (FBe) on loperamide (LP)-induced constipation in rats. BMC Complement. Altern. Med. 19(1), 143.

Lizuka, N. and Hamamoto, Y. 2015. Constipation and herbal medicine. Front. Pharmacol. 6, 73.

Najafian, Y., Hamedi, S.S., Kaboli Farshchi, M. and Feyzabadi, Z. 2018. Plantago major in traditional persian medicine and modern phytotherapy: a narrative review. Electron. Physician. 10(2), 6390–6399.

Panche, A.N., Diwan, A.D. and Chandra, S.R. 2016. Flavonoids: an overview. J. Nutr. Sci. 5, 47.

Papale, A.J., Flattau, R., Vithlani, N., Mahajan, D. and Nadella, S. 2024. A review of pharmacologic and non-pharmacologic therapies in the management of irritable bowel syndrome: current recommendations and evidence. J. Med. Clin. 13(22), 6948.

Parkar, N., Spencer, N.J., Wiklendt, L., Olson, T., Young, W., Janssen, P., McNabb, W.C. and Dalziel, J.E. 2024. Novel insights into mechanisms of inhibition of colonic motility by loperamide. Front. Neurosci. 18, 1424936.

Saptarini, N.M., Mustarichie, R., Hasanuddin, S. and Corpuz, M.J.T. 2024. Cassia alata L.: a study of antifungal activity against malassezia furfur, identification of major compounds, and molecular docking to lanosterol 14-alpha demethylase. Pharmaceuticals (Basel) 17(3), 380.

Stevick, R.J., Audrain, B., Bedu, S., Dray, N., Ghigo, J.M. and Pérez-Pascual, D. 2023. Anti-diarrheal drug loperamide induces dysbiosis in zebrafish microbiota via bacterial inhibition. Microbiome 11(1), 252.

Włodarczyk, J., Waśniewska, A., Fichna, J., Dziki, A., Dziki, Ł. and Włodarczyk, M. 2021. Current overview on clinical management of chronic constipation. J. Clin. Med. 10(8), 1738.

Wresdiyati, T., Sa’diah, S., Astawan, M., Alfarisi, H., Aziz, S.A., Darawati, M. and Subangkit, M. 2023. The repeated dose 28-day oral toxicity study of combined extract of Cajanus cajan leaf and Zingiber officinale rhizome in male and female Sprague-Dawley rats. Trop. J. Nat. Prod. Res. 7(8), 3706–3716.

Zhang, S., Hu, J., Sun, Y., Tan, H., Yin, J., Geng, F. and Nie, S. 2021. Review of structure and bioactivity of the Plantago (Plantaginaceae) polysaccharides. Food. Chem. X. 12, 100158.

Zhao, L. and Zheng, L. 2023. A review on bioactive anthraquinone and derivatives as the regulators for ROS. Molecules 28(4), 8139.

Zhao, Q., Chen, Y.Y., Xu, D.Q., Yue, S.J., Fu, R.J., Yang, J., Xing, L.M. and Tang, Y.P. 2021. Action mode of gut motility, fluid and electrolyte transport in chronic constipation. Front. Pharmacol. 12, 630249.