Pharmacological basis for medicinal use of Cymbopogon proximus Hochst. ex A. Rich. Essential oil in hyperactive gastrointestinal disorders

Abstract

Background:

Cymbopogon is used in traditional medicine in tropical and subtropical regions to treat various ailments. However, no research has yet provided a detailed pharmacodynamic explanation for its antidiarrheal and antispasmodic effects. This study aimed to evaluate the antidiarrheal and antispasmodic properties of the essential oil of Cymbopogon proximus (EOCP), with the goal of scientifically validating its traditional use in folk medicine.

Methods:

An animal study of antidiarrheal activity was carried out using a castor oil-induced diarrhea model in rats, and the isolated small intestine of rats was used to investigate the specific mechanisms of the antispasmodic effects.

Results:

The EOCP, exhibited a protective effect against castor oil-induced diarrhea in rats at 100 and 200 mg/kg, similar to the standard drug, dicyclomine. In rat ileum preparations, EOCP decreased the basal tonus with a maximal response of (R_max; % of ACh.-contraction) of 18.5% ± 1.5% compared with 17.5% ± 2.5% achieved with dicyclomine. In contractions elicited by different agents, EOCP showed higher potency in relaxing carbachol (CCh)-induced contractions compared to those induced by high K⁺ concentrations. It showed a similar relaxation profile to dicyclomine but differed from the inhibitory effects of verapamil and/or atropine. Pre-incubation of isolated ileum with increasing doses of EOCP showed that at a lower concentration (0.03 mg/mL), EOCP induced a rightward parallel shift in carbachol (CCh) concentration-response curves. At a higher concentration (0.1 mg/mL), it caused a non-parallel shift with a reduction in the maximum response, similar to the effects of dicyclomine, a dual inhibitor of muscarinic receptors and Ca⁺⁺ channels. The Ca⁺⁺ channel inhibitory effect of EOCP was confirmed by its rightward shift of Ca⁺⁺ concentration-response curves and suppression of the maximal response, resembling the effects of verapamil, a well-established Ca⁺⁺ antagonist.

Conclusion:

These results suggest that EOCP produces antidiarrheal and antispasmodic effects, possibly mediated via direct effect on intestinal smooth musclesf followed by dual inhibition of muscarinic receptors and Ca⁺⁺ channels, thus providing a pharmacological basis for its traditional use in hyperactive gut disorders such as diarrhea and spasms.

1 Introduction

Cymbopogon, known for its rich essential oil content, has long been used in traditional medicine across the tropical and subtropical regions of Asia, Africa, and the Americas (Avoseh et al., 2015). Its medicinal uses include treatment of ailments such as cough, fever, infections, cancer, and digestive disorders (Dutta et al., 2016). Cymbopogon lowers blood pressure in normotensive rats and provides protection against L-NAME-induced hypertension (El-Nezhawy et al., 2014; Althurwi et al., 2020). Laboratory and animal studies demonstrate its pharmacological benefits, including anticancer, heart-protective, cholesterol-lowering, antioxidant, anti-inflammatory, anti-diabetic, and antimicrobial effects (Mansour et al., 2002; Adeneye and Agbaje, 2007; Miguel, 2010; Selim, 2011; Ekpenyong et al., 2014).

Cymbopogon proximus (commonly known as Halfabar or Maharaib) is a highly aromatic grass commonly found in Southern Egypt and Northern Sudan. C. proximus is traditionally used as a diuretic and antispasmodic, which is attributed to its potent smooth muscle-relaxing effects (El-Askary et al., 2003). Additionally, it possesses various biological activities, including hypoglycemic, antipyretic, bronchodilatory, antibacterial, anticonvulsant, and antiemetic properties (El-Askary et al., 2003; Selim, 2011; Ibrahim and El-Khateeb, 2013; El-Nezhawy et al., 2014; Warrag et al., 2014). In our earlier conducted study on the essential oil of this plant (Althurwi et al., 2020), the detailed phytochemical GC-MS analysis revealed the presence of elemol, piperitone, α-eudesmol and β-eudesmol in as the plant major constituents. Interestingly, some of the identified components in C. proximus are known to have spasmolytic properties such as, piperitone, with previously reported findings of having concentration-dependent spasmolytic activity (Ponce-Monter et al., 2008). Moreover, β-eudesmol, found in C. Proximus essential oil, has been explored for its relaxing effect in previous studies (Morita et al., 1996). Although its antispasmodic and smooth muscle relaxant effects have been established in previous studies (Abdel-Moneim et al., 1969), no detailed pharmacodynamic explanation for its antidiarrheal and antispasmodic effects has been provided. Therefore, the current study aimed to scientifically investigate the antidiarrheal and antispasmodic properties of the essential oil of Cymbopogon proximus (EOCP), to provide a pharmacological basis for its traditional use in the treatment of hyperactive gut disorders like diarrhea and spasms. While Cymbopogon proximus has long been used in traditional medicine, no previous research has provided a clear mechanistic explanation for its effectiveness in treating hyperactive gut disorders. This study identifies EOCP’s dual inhibitory effect on muscarinic receptors and calcium channels and positions it as a natural alternative to established pharmaceuticals such as dicyclomine. The broader significance of these findings extends to the potential development of EOCP as a plant-based therapeutic for gastrointestinal conditions, offering a safer, natural remedy at a time of increasing interest in alternative and complementary medicine. Additionally, this research could open avenues for further exploration of other Cymbopogon species and their medicinal properties, thereby contributing to the advancement of ethnopharmacology and drug development from natural products.

2 Materials and methods

2.1 Plant material and extraction

Cymbopogon proximus belonging to the Poaceae family, was sourced from a local market in Alexandria, Egypt. The plant’s identity was confirmed by Prof. Saniya Kamal of the Department of Botany, College of Science, Alexandria University, Alexandria, Egypt.

2.2 Extraction of the essential oil Cymbopogon proximus

Essential oil was extracted from 250 g of dried, powdered C. proximus by hydrodistillation for 5 h of (Meyer-Warnod, 1984). The oil was then separated and dried with anhydrous sodium sulfate, giving a final yield of 5.4% w/w (Althurwi et al., 2020).

2.3 Chemicals

Acetylcholine chloride (ACh), atropine, carbachol (CCh), dicyclomine, Tween 80 and verapamil were sourced from Sigma Chemicals Co. (St. Louis, MO, United States). The physiological salt solutions were prepared using potassium chloride (Sigma Chemicals Co.), calcium chloride, magnesium chloride, magnesium sulfate, glucose, sodium bicarbonate, potassium dihydrogen phosphate, and sodium dihydrogen phosphate (Merck, Darmstadt, Germany), ethylenediaminetetraacetic acid (EDTA), and sodium chloride (BDH Laboratory Supplies, Poole, England). All chemicals used were of analytical grade. Castor oil was obtained from a local pharmacy in Al-Kharj. EOCP was administered in a volume of 10 mL/kg after dissolving in a vehicle of 1% (w/v) Tween 80 in saline in the in-vivo assay whereas for the ex-vivo studies, EOCP stock solution and dilutions were prepared in Tyrode’s solution. The prepared stock solutions of EOCP were sonicated just before use.

2.4 Animals

Male Wistar albino rats, weighing 200–250 g, were obtained from the Lab Animal Care Unit of the College of Pharmacy, Prince Sattam Bin Abdulaziz University (Al-Kharj, KSA). The animals were housed under a 12-h light/dark cycle and received food and water ad libitum. After a 24-h fasting period, the rats were humanely euthanized by a blow to the head, followed by cervical dislocation. All experiments were conducted in accordance with the guidelines set by the Institute of Laboratory Animal Resources, Commission on Life Sciences, National Research Council (1996). The assay protocol was approved by the Standing Committee of Bioethics Research (SCBR) at Prince Sattam Bin Abdulaziz University with reference number SCBR-394/2024.

2.5 Castor oil-induced diarrhea

Rats were subjected to 24 h fasting and kept individually in separate cages with saw dust replaced with a clean blotting sheet. They were divided into five groups, each consisting of five rats. Each rat in the first group received saline (1% Tween 80 at 10 mL/kg, orally) and was recorded as a negative control. Effective doses of plant oil were previously determined on a trial basis, with the second and third groups receiving two increasing doses of EOCP as 100 and 200 mg/kg previously dissolved in vehicle (1% Tween 80 at 10 mL/kg, orally). The fourth and fifth groups received the positive control drug, dicyclomine, at oral doses of 50 and 100 mg/kg, respectively. Each rat was carefully subjected to 10 mL/kg dose of castor oil after approximately 1 h of sample treatment. After 4 h of the castor oil dosing, the cages were subsequently monitored for loose spots of diarrhea, and their absence was recorded as a positive result, indicating protection from diarrhea (Rehman et al., 2012).

2.6 Rat ileum

The rat’s abdomen was opened, and with due care, the ileum was isolated by tracing it from the ileo-cecal junction, placed in a normal buffer (Tyrode’s solution), and mesenteries were removed (Rehman et al., 2024). A 2-cm segment of the ileum was suspended in a 10 mL tissue bath filled with Tyrode’s solution (pH 7.4), maintained at 37°C, and aerated with a mixture of 95% O₂ and 5% CO₂ (carbogen). The composition of the Tyrode’s solution (in mM) was as follows: NaCl 136.9, KCl 2.7, MgCl₂·6H₂O 0.5, NaHCO₃ 11.9, NaH₂PO₄·2H₂O 0.32, CaCl₂ 1.8, and glucose 5.05. One end of the ileum was secured to a metal tissue hook, while the other was attached via a cotton thread to an isotonic transducer connected to an emkaBath (France) system. Tissue responses were recorded using IOX software (version 2.9.10.6, emka technologies, SAS, Paris, France). Each tissue segment was subjected to an initial load of 1 g and allowed to equilibrate for 30 min before drugs were administered. After the equilibration phase, each tissue preparation was stabilized using a submaximal concentration of ACh, (0.3 μM) at 3-min intervals, until stable responses were observed. The inhibitory effect of the test substances was evaluated against contractions induced by CCh (1 μM) and high potassium (80 mM). Carbachol, a muscarinic receptor agonist, induces strong contractions in isolated ileum tissue, and any substance that reverses these contractions is considered anti-muscarinic (Arunlakshana and Schild, 1997).

To confirm whether the inhibitory substance acts competitively or non-competitively on muscarinic receptors, control concentration-response-curves (CRCs) for CCh-were generated by incrementally increasing the CCh concentration. CRCs for CCh were repeated with increasing concentrations of the test substance (EOCP) and control drugs (dicyclomine, verapamil, and atropine), as described by Choo and Mitchelson (1978).

For determining the calcium channel-blocking signaling, high potassium (80-mM) was used to contract the tissue sample, as outlined by Farre et al. (1991). Potassium concentrations (>30 mM) have been reported to cause smooth muscle depolarization by activating membrane voltage-dependent calcium signaling channels, thus facilitating the entry of extracellular calcium and triggering contractions. A substance that inhibits these contractions is considered a blocker of calcium influx through L-type calcium channels (Godfraind et al., 1986). Once the contraction reached a plateau (usually within 7–10 min), the test material was added cumulatively to achieve concentration-dependent inhibitory effects.

To further verify the calcium antagonist activity of the test substance, the tissue was first stabilized in normal Tyrode’s solution. This was then replaced with a calcium-free Tyrode’s solution containing 0.1 mM EDTA for 30 min to remove calcium from the tissue. The solution was then exchanged for a potassium-rich, calcium-free Tyrode’s solution with the following composition (in mM): NaCl 91.03, KCl 50, MgCl₂·6H2O 0.50, NaHCO₃ 11.9, NaH₂PO₄·2H₂O 0.32, glucose 5.05, and EDTA-Na₂·2H₂O 0.1. After incubating for 30 min, control calcium concentration-response curves (CRCs) were generated. Once the calcium CRCs had stabilized (typically after two cycles), the tissue was exposed to the test compound for 1 h. Calcium CRCs were then reconstructed at varying concentrations of the test substance to evaluate their calcium antagonist effect.

2.7 Statistical analyses

Data are expressed as mean ± standard error of the mean (SEM, n = number of experiments), along with median effective concentrations (EC₅₀) and corresponding 95% confidence intervals (CI). Statistical analysis for the antidiarrheal assay was performed using the Chi-square test, while potency of the agonist control CRCs) was compared with its potency in the presence of the different antagonist(s) using Oneway ANOVA and Dunnett’s multiple comparison test. Repeated Measures ANOVA followed by Bonferroni’s post-test for comparisons of CRCs of Ca⁺⁺ with their respective controls. Differences were considered statistically significant at P < 0.05.

Concentration-response curves were evaluated using non-linear regression analysis with GraphPad software (GraphPAD, San Diego, CA, United States).

3 Results

3.1 Effect on castor oil-induced diarrhea

The tested oil (EOCP) exhibited dose-dependent protective effects (40% and 80%) against castor oil-induced diarrhea in rats at 100 and 200 mg/kg, respectively. All rats of the negative control group (saline treated) were recorded with loose diarrheal spots and therefore did not exhibit protection against castor oil-induced diarrhea. However, the positive control drug, dicyclomine, showed 40% protection at a lower dose of 50 mg/kg, while its higher dose (100 mg/kg) showed complete (100%) protection (Table 1).

3.2 Relaxant effect on the baseline of rat ileum

Figures 1A–D shows the original representative tracings of the effect of EOCP, dicyclomine, verapamil and atropine on the baseline contraction of the rat isolated ileum. The essential oil of C. proxismus was shown to have a direct relaxant effect on the baseline intestinal tone (Figure 1A) with a maximum relaxant response (R_max) corresponded to 18.5% ± 1.5% of the ACh (0.3 µM)-induced contraction (Figure 1E). This effect of EOCP was equivalent to the maximal relaxant response induced by dicyclomine (Figure 1B) which was recorded with R_max of 17.5% ± 2.5% (Figure 1E) whereas verapamil (Figure 1C) and atropine (Figure 1D) relaxed only slightly the baseline contraction of ileum tissue with resultant R_max of 3.5% ± 1.5% and 4.5% ± 2.5%, respectively (Figure 1E).

FIGURE 1

3.3 Antispasmodic effect on the induced contractoins of rat ileum

Figure 2 shows the representative concentration-dependent antispasmodic effect of the EOCP on high K⁺-induced contractions. In both, CCh (1 µM) and K⁺ (80 mM)-induced contractions, complete relaxation was observed by EOCP. However, EOCP reversed CCh-induced contractions at lower concentrations, with a recorded EC₅₀ of 0.12 mg/mL (0.10–0.15), n = 5, demonstrating higher potency compared to its inhibitory effect against K⁺-induced contractions where lower potency was observed with resultant EC₅₀ of 1.15 mg/mL (1.07–1.28), n = 5, as shown in Table 2 and Figure 3A. Dicyclomine, also showed a similar pattern of inhibitory activity against CCh- and K⁺-induced contractions with corresponding EC₅₀ values of 0.31 μM (0.24–0.41), n = 5 and 3.41 μM (3.01–3.86), n = 5 (Figure 3B). However, verapamil exhibited greater potency against K⁺-induced contractions, with EC₅₀ of 0.11 μM (0.08–0.13), n = 5, and its effect on CCh-induced contractions, had an EC₅₀ 0.92 μM (0.83–1.01), n = 5, Figure 3C. Control drug, atropine effectively counteracted contractions induced by CCh (1 µM) with an EC₅₀ of 0.29 μM (0.26–0.32), n = 5, without any effect on K⁺-induced contractions (Figure 3D). The detailed EC₅₀ values for EOCP and control drugs are shown in Table 2.

FIGURE 2

FIGURE 3

3.4 Anticholinergic activity confirmation

At a lower concentration of 0.03 mg/mL, EOCP caused a rightward parallel shift in the CCh-CRCs without reducing the maximum contractile response of the agonist, followed by a non-parallel shift with suppression of the maximal response at a higher concentration of 0.1 mg/mL (Figure 4A; Table 3). Similarly, dicyclomine (0.01 and 0.03 µM) showed a similar shift pattern by showing suppression of the maximum response only at higher concentration of 0.03 µM where the maximum contractile response of CCh was recorded as 56.6% whereas its lower concentration significantly effected CCh potency (p < 0.01) without decreasing the maximum response of CCh (Table 3; Figure 4B). Pre-incubation with atropine (0.01 and 0.03 µM) caused a rightward parallel shift in the CCh-CRCs without suppressing the maximal effect while the potency of CCh was significantly effected (p < 0.01) at both concentrations (Table 3; Figure 4C). In contrast, verapamil (0.01 and 0.03 µM) caused a non-parallel rightward shift in CCh-CRCs with a decrease in the maximum response of control curves to 79% and 51.3% at verapamil pre-incubated concentration of 0.01 µM and 0.03 µM, respectively (Table 3; Figure 4D).

FIGURE 4

3.5 Ca⁺⁺ channel blocking activity confirmation

When tested for possible interactions with Ca⁺⁺ channels, EOCP (0.1 and 0.3 mg/mL) induced a rightward shift of Ca⁺⁺ concentration-response curves (CRCs) and suppressed the maximal response (Figure 5A), mirroring the effect of verapamil (Figure 5B) and dicyclomine (Figure 5C).

FIGURE 5

4 Discussion

Given the known medicinal use of C. proximus as an antispasmodic (El-Askary et al., 2003) and its smooth muscle relaxant properties (Abdel-Moneim et al., 1969), the extracted essential oil was tested on laboratory rats to evaluate its potential effects on hyperactive gut disorders, such as diarrhea and spasms. In a castor oil-induced diarrhea model, EOCP demonstrated a dose-dependent protective effect similar to dicyclomine, a standard antidiarrheal agent (Pasricha, 2006). Castor oil causes diarrhea due to the ricinoleic acid, formed during hydrolysis, which disrupts electrolyte and water transport, and leads to strong intestinal contractions (Iwao and Terada, 1962; Croci et al., 1997). Therefore, an effective antidiarrheal remedy can work by inhibiting bowel contractions.

Essential oil of C. proxismus had a direct relaxant effect on the isolated ileum preparation of rat and also relaxed tissue with induced contractions by different spasmodic agents. These effects were observed at lower concentrations and might be related to the symptomatic relief of the acute abdominal pain associated with some gastric disturbances (Câmara et al., 2003). Next, to explore the mechanism underlying EOCP’s antidiarrheal effects, its activity was tested against contractions induced by CCh and high potassium (K⁺). Medicinal plants extracts or thier extracted essential oils usually possess antispasmodic effect on multiple smooth muscles including intestinal smooth muscles, possibly mediated by combination of mechanism(s) such as anticholinergic (Mehmood et al., 2011), Ca⁺⁺ channel blockade (Gilani et al., 2010; Rehman et al., 2021), phosphodiesterase inhibition (Arshad et al., 2016; Imam et al., 2020) and/or potassium channel activation (Ansari et al., 2024; Khan et al., 2011). Notably, EOCP reversed CCh-induced contractions more effectively than those caused by high K⁺. Dicyclomine, which blocks both muscarinic receptors and calcium (Ca⁺⁺) influx (McGrath et al., 1964; Downie et al., 1977), displayed a similar inhibition pattern, whereas verapamil, a known Ca⁺⁺ signaling inhibitor (Jenkinson, 2002), was observed to be potent for reversing high K⁺-provoked spasms than those induced by CCh. Atropine, a muscarinic receptor non-specific blocker (Arunlakshana and Schild, 1997), only relaxed CCh-induced contractions. These results suggested that EOCP has both muscarinic receptor blocking and Ca⁺⁺ influx inhibitory effects.

Further confirmation of EOCP’s effects was obtained by constructing CCh and Ca⁺⁺ CRCs in the presence of different concentrations of tested samples. At lower doses, EOCP caused a parallel shift in CCh curves without reducing the maximal effect, indicating competitive inhibition, similar to atropine (Eglen and Harris, 1993; Jenkinson, 2002). At higher concentrations, EOCP produced a non-parallel deflection with a reduced peak effect previously observed in control curves, suggesting non-competitive inhibition, as observed with Ca⁺⁺ antagonists (Van den Brink, 1973; Irie et al., 2000). Dicyclomine also deflected the CCh curves in a pattern similar to EOCP, while verapamil caused a rightward, non-parallel manner shift of curves with a reduction in the maximum peaks at all pre-incubated concentrations. In contrast, atropine caused a parallel rightward shift without reducing the maximum response. EOCP shifted the Ca⁺⁺ curves to the right and reduced the maximal response, akin to the effects of verapamil and dicyclomine. Our findings align with the study by Marghich et al. (2023), which demonstrated that the myorelaxant and antispasmodic effects of Artemisia campestris essential oil are likely mediated through conformational changes in cholinergic muscarinic receptors and inhibition of L-type voltage-gated calcium channels, with a predominant role attributed to the former mechanism. These findings were further supported by molecular docking analyses. Similarly, several studies on medicinal plants with antidiarrheal and antispasmodic properties (Aleem and Janbaz, 2018; Mehmood et al., 2011) have reported smooth muscle relaxant mechanisms comparable to those observed for EOCP, reinforcing the validity of our current results.

Cholinergic antagonists are well-established therapeutic agents for the treatment of diarrhea (Pietrusko, 1979); however, their use is often associated with adverse cardiac stimulation, particularly when administered orally (Nawarth, 1981). Conversely, calcium channel blockers (CCBs) are also effective in managing hypermotile gut conditions (Borsari, 1991) but are known for their cardio-suppressant effects (Billman, 1992). The presence of CCB-like constituents alongside antimuscarinic compounds in Cymbopogon proximus essential oil may represent a naturally occurring mechanism designed to counteract tachycardia, a common side effect of standalone anticholinergic agents. This phenomenon aligns with the concept that natural remedies often exhibit “effect-enhancing and/or side-effect-neutralizing” properties (Gilani and Rahman, 2005), in addition to their cost-effectiveness and potential merit in evidence-based medicine (Ernst, 2005).

This study builds on the traditional use of Cymbopogon species as natural remedies to treat gastrointestinal disorders, such as diarrhea and spasms, and supports their historical use in folk medicine (Zhao et al., 2024). For instance, Cymbopogon citratus essential oil and its major component, citral, exhibit strong antidiarrheal effects by reducing intestinal motility and mitigating gut hyperactivity (Tangpu and Yadav, 2006). Additionally, Cymbopogon martinii essential oil exhibits spasmolytic properties by inhibiting spontaneous contractions in the isolated rabbit jejunum and suppressing potassium-induced contractions, functioning similarly to calcium channel blockers such as verapamil (Janbaz et al., 2014). Research also revealed that Cymbopogon schoenanthus essential oil from Sudan has remarkable spasmolytic activity (Pavlović et al., 2017; Djemam et al., 2020).

The effects observed in C. proximus may be attributed to the rich composition of its essential oils (Al-Taweel et al., 2013; Althurwi et al., 2020). Notably, some of the identified components in C. proximus are known to have spasmolytic properties. For example, Piperitone, a major metabolite in the essential oil studied, was previously shown to have concentration-dependent spasmolytic activity (Ponce-Monter et al., 2008). Additionally, β-eudesmol, another metabolite of C. Proximus essential oil, has shown relaxing effect in previous studies (Morita et al., 1996). Limonene, another minor component of this oil, has also been proven to have spasmolytic activity (Cardoso Lima et al., 2012).

This study provides valuable insight into the dual mechanism of C. proximus essential oil (EOCP) as an antidiarrheal and antispasmodic agent, but several limitations should be acknowledged. The results based on animal models, may not be directly applicable to humans, and the study lacks examination of other relevant physiological pathways and detailed pharmacokinetic data. Additionally, more research is needed to identify the key components responsible for the observed effects. However, previous studies have documented the antispasmodic effects of C. Proximus. This study introduces novel insights by providing a more detailed mechanistic understanding, showing that EOCP acts through both muscarinic receptor inhibition and calcium influx blockade.

5 Conclusion

These results suggest that the essential oil extracted from C. proximus, independently, had a direct relaxant and spasmolytic effect on the intestinal smooth muscles that does not depend on interaction with receptors for neurotransmitters and are probably myogenic in nature. Moreover, the antidiarrheal and antispasmodic effects of EOCP is to a great extent may be mediated by the dual inhibition of muscarinic receptors and Ca⁺⁺ channels. However, the possible involvement of additional inhibitory mechanism(s) cannot be ruled out. Thus, this study provides a scientific rationale for the traditional use of C. proximus in treating gut hyperactivity disorders such as diarrhea and spasms, while paving the way for future studies to explore its potential for broader therapeutic applications. Future research should also examine its long-term safety, effectiveness, and the molecular mechanisms underlying its actions.

References

• 1

  Abdel-Moneim F. M. Ahmed Z. F. Fayez M. B. E. Ghaleb H. (1969). Constituents of local plants. XIV. The antispasmodic principle in Cymbopogon proximus. Planta Med.17, 209–216. 10.1055/s-0028-1099848

  • CrossRef
  • Google Scholar

• 2

  Adeneye A. A. Agbaje E. O. (2007). Hypoglycemic and hypolipidemic effects of fresh leaf aqueous extract of Cymbopogon citratus stapf. in rats. J. Ethnopharmacol.112, 440–444. 10.1016/j.jep.2007.03.034

  • CrossRef
  • Google Scholar

• 3

  Aleem A. Janbaz K. H. (2018). Dual mechanisms of anti-muscarinic and Ca⁺⁺ antagonistic activities to validate the folkloric uses of Cyperus niveus retz. as antispasmodic and antidiarrheal. J. Ethnopharmacol.1, 138–148. 10.1016/j.jep.2017.11.006

  • CrossRef
  • Google Scholar

• 4

  Al-Taweel A. M. Fawzy G. A. Perveen S. El Tahir K. E. H. (2013). Gas chromatographic mass analysis and further pharmacological actions of Cymbopogon proximus essential oil. Drug Res. Stuttg.63, 484–488. 10.1055/s-0033-1347239

  • CrossRef
  • Google Scholar

• 5

  Althurwi H. N. Abdel-Kader M. S. Alharthy K. M. Salkini M. A. Albaqami F. F. (2020). Cymbopogon proximus essential oil protects rats against isoproterenol-induced cardiac hypertrophy and fibrosis. Molecules25, 1786. 10.3390/molecules25081786

  • CrossRef
  • Google Scholar

• 6

  Ansari M. N. Rehman N. U. Samad A. Ahmad W. (2024). Pharmacological basis for the antidiarrheal and antispasmodic effects of cuminaldehyde in experimental animals: in silico, in vitro and in vivo studies. Front. Biosci.29, 43. 10.31083/j.fbl2901043

  • CrossRef
  • Google Scholar

• 7

  Arshad U. Bashir S. Rehman N. U. Yaqub T. Gilani A. H. (2016). Dual inhibition of Ca+2 influx and phosphodiesterase enzyme provides scientific base for the medicinal use of Chrozophora prostrata dalz. In respiratory disorders. Phytother. Res.30, 1010–1015. 10.1002/ptr.5610

  • CrossRef
  • Google Scholar

• 8

  Arunlakshana O. Schild H. O. (1997). Some quantitative uses of drug antagonists. 1958. Br. J. Pharmacol.120 (Suppl. ment), 151–161. 10.1111/j.1476-5381.1997.tb06793.x

  • CrossRef
  • Google Scholar

• 9

  Avoseh O. Oyedeji O. Rungqu P. Nkeh-Chungag B. Oyedeji A. (2015). Cymbopogon species; ethnopharmacology, phytochemistry and the pharmacological importance. Molecules20, 7438–7453. 10.3390/molecules20057438

  • CrossRef
  • Google Scholar

• 10

  Billman G. E. (1992). “The antiarrythmic effects of the calcium antagonists,” in Calcium antagonists in clinical medicine. Editor EpsteinM. (Philadelphia, United States: Hanley and Belfus Inc.), 183–212.

  • Google Scholar

• 11

  Borsari G. (1991). L'effet thérapeutique de l'anticalcique vérapamil dans la diarrhée chronique. Communication préliminaire [The therapeutic effects of the calcium blocker verapamil in chronic diarrhea. Preliminary communication]. Schweiz. Med. wochenschr.31, 1238–1242.

  • Google Scholar

• 12

  Câmara C. C. Nascimento N. R. Macêdo-Filho C. L. Almeida F. B. Fonteles M. C. (2003). Antispasmodic effect of the essential oil of Plectranthus barbatus and some major constituents on the guinea-pig ileum. Planta Med.69, 1080–1085. 10.1055/s-2003-45186

  • CrossRef
  • Google Scholar

• 13

  Cardoso Lima T. Mota M. M. Barbosa-Filho J. M. Viana Dos Santos M. R. De Sousa D. P. (2012). Structural relationships and vasorelaxant activity of monoterpenes. Daru20, 23. 10.1186/2008-2231-20-23

  • CrossRef
  • Google Scholar

• 14

  Choo L. K. Mitchelson F. (1978). Antagonism of cholinomimetics by troxypyrrolidinium in guinea-pig atria and longitudinal ileal muscle: Comparison with hemicholinium-3. Eur. J. Pharmacol.52, 313–322. 10.1016/0014-2999(78)90284-4

  • CrossRef
  • Google Scholar

• 15

  Croci T. Landi M. Emonds-Alt X. Le Fur G. Maffrand J. P. Manara L. (1997). Role of tachykinins in Castor oil diarrhoea in rats. Br. J. Pharmacol.121, 375–380. 10.1038/sj.bjp.0701130

  • CrossRef
  • Google Scholar

• 16

  Djemam N. Lassed S. Gül F. Altun M. Monteiro M. Menezes-Pinto D. et al (2020). Characterization of ethyl acetate and n-butanol extracts of Cymbopogon Schoenanthus and Helianthemum lippii and their effect on the smooth muscle of the rat distal colon. J. Ethnopharmacol.252, 112613. 10.1016/j.jep.2020.112613

  • CrossRef
  • Google Scholar

• 17

  Downie J. W. Twiddy D. A. Awad S. A. (1977). Antimuscarinic and noncompetitive antagonist properties of dicyclomine hydrochloride in isolated human and rabbit bladder muscle. J. Pharmacol. Exp. Ther.201, 662–668. 10.1016/s0022-3565(25)30905-5

  • CrossRef
  • Google Scholar

• 18

  Dutta S. Munda S. Lal M. Bhattacharyya P. R. (2016). A short review on chemical composition therapeutic use and enzyme inhibition activities of cymbopogon species. Indian J. Sci. Technol.9, 1–9. 10.17485/ijst/2016/v9i46/87046

  • CrossRef
  • Google Scholar

• 19

  Eglen R. M. Harris G. C. (1993). Selective inactivation of muscarinic M2 and M3 receptors in guinea-pig ileum and atria in vitro. Br. J. Pharmacol.109, 946–952. 10.1111/j.1476-5381.1993.tb13712.x

  • CrossRef
  • Google Scholar

• 20

  Ekpenyong C. E. Davies K. Antai E. E. (2014). Cymbopogon citratus Stapf (DC) extract ameliorates atherogenic cardiovascular risk in diabetes-induced dyslipidemia in rats. Br. J. Med. Med. Res.4, 4695–4709. 10.9734/BJMMR/2014/11262

  • CrossRef
  • Google Scholar

• 21

  El-Askary H. I. Meselhy M. R. Galal A. M. (2003). Sesquiterpenes from Cymbopogon proximus. Molecules8, 670–677. 10.3390/80900670

  • CrossRef
  • Google Scholar

• 22

  El-Nezhawy A. O. H. Maghrabi I. A. Mohamed K. M. Omar H. A. (2014). Cymbopogon proximus extract decreases L-NAME-induced hypertension in rats. Int. J. Pharm. Sci. Rev. Res.27, 66–69.

  • Google Scholar

• 23

  Ernst E. (2005). The efficacy of herbal medicine-an overview. Fundam. Clin. Pharmacol.19, 405–409. 10.1111/j.1472-8206.2005.00335.x

  • CrossRef
  • Google Scholar

• 24

  Farre A. J. Colombo M. Fort M. Gutierrez B. (1991). Differential effects of various Ca²⁺ antagonists. Gen. Pharmacol.22, 177–181. 10.1016/0306-3623(91)90331-y

  • CrossRef
  • Google Scholar

• 25

  Gilani A. H. Mandukhail S. U. Iqbal J. Yasinzai M. Aziz N. Khan A. et al (2010). Antispasmodic and vasodilator activities of Morinda citrifolia root extract are mediated through blockade of voltage dependent calcium channels. Bmc. Complement. Altern. Med.10, 2. 10.1186/1472-6882-10-2

  • CrossRef
  • Google Scholar

• 26

  Gilani A. H. Rahman A. (2005). Trends in ethnopharmocology. J. Ethnopharmacol.100, 43–49. 10.1016/j.jep.2005.06.001

  • CrossRef
  • Google Scholar

• 27

  Godfraind T. Miller R. Wibo M. (1986). Calcium antagonism and calcium entry blockade. Pharmacol. Rev.38, 321–416. 10.1016/s0031-6997(25)06867-x

  • CrossRef
  • Google Scholar

• 28

  Ibrahim F. Y. El-Khateeb A. Y. (2013). Effect of herbal beverages of Foeniculum vulgare and Cymbopogon proximus on inhibition of calcium oxalate renal crystals formation in rats. Ann. Agric. Sci.58, 221–229. 10.1016/j.aoas.2013.07.006

  • CrossRef
  • Google Scholar

• 29

  Imam F. Rehman N. U. Ansari M. N. Qamar W. Afzal M. Alharbi K. S. (2020). Effect of roflumilast in airways disorders via dual inhibition of phosphodiesterase and Ca²⁺-channel. Saudi. Pharm. J.28, 698–702. 10.1016/j.jsps.2020.04.011

  • CrossRef
  • Google Scholar

• 30

  Irie K. Yoshioka T. Nakai A. Ochiai K. Nishikori T. Wu G. R. et al (2000). A Ca(2+) channel blocker-like effect of dehydrocurdione on rodent intestinal and vascular smooth muscle. Eur. J. Pharmacol.403, 235–242. 10.1016/s0014-2999(00)00445-3

  • CrossRef
  • Google Scholar

• 31

  Iwao I. Terada Y. (1962). On the mechanism of diarrhea due to Castor oil. Jpn. J. Pharmacol.12, 137–145. 10.1254/jjp.12.137

  • CrossRef
  • Google Scholar

• 32

  Janbaz K. H. Qayyum A. Saqib F. Imran I. Zia-Ul-Haq M. De Feo V. (2014). Bronchodilator, vasodilator and spasmolytic activities of Cymbopogon martinii. J. Physiol. Pharmacol.65, 859–866.

  • Google Scholar

• 33

  Jenkinson D. H. (2002). “Classical approaches to the study of drug-receptor interactions,” in Textbook of receptor pharmacology, Editors Foreman JC, Johansen T, (Boca Raton, FL: CRC Press), 3–78.

  • Google Scholar

• 34

  Khan A. Rehman N. U. AlKharfy K. M. Gilani A. H. (2011). Antidiarrheal and antispasmodic activities of Salvia officinalis are mediated through activation of K+ channels. Bang. J. Pharmacol.6, 111–116. 10.3329/bjp.v6i2.9156

  • CrossRef
  • Google Scholar

• 35

  Mansour H. A. Newairy A.-S. A. Yousef M. I. Sheweita S. A. (2002). Biochemical study on the effects of some Egyptian herbs in alloxan-induced diabetic rats. Toxicology170, 221–228. 10.1016/s0300-483x(01)00555-8

  • CrossRef
  • Google Scholar

• 36

  Marghich M. Amrani O. Karim A. Harit T. Beyi L. Mekhfi H. et al (2023). Myorelaxant and antispasmodic effects of the essential oil of Artemisia campestris L., and the molecular docking of its major constituents with the muscarinic receptor and the L-type voltage-gated Ca²⁺ channel. J. Ethnopharmacol.15, 116456. 10.1016/j.jep.2023.116456

  • CrossRef
  • Google Scholar

• 37

  McGrath W. R. Lewis R. E. Kuhn W. L. (1964). The dual mode of the antispasmodic effect of dicyclomine hydrochloride. J. Pharmacol. Exp. Ther.146, 354–358. 10.1016/s0022-3565(25)26933-6

  • CrossRef
  • Google Scholar

• 38

  Mehmood M. H. Siddiqi H. S. Gilani A. H. (2011). The antidiarrheal and spasmolytic activities of Phyllanthus emblica are mediated through dual blockade of muscarinic receptors and Ca2+ channels. J. Ethnopharmacol.27, 856–865. 10.1016/j.jep.2010.11.023

  • CrossRef
  • Google Scholar

• 39

  Meyer-Warnod B. (1984). Natural essential oils: extraction processes and applications to some major oils. Perfum. Flavorist, 93–103.

  • Google Scholar

• 40

  Miguel M. G. (2010). Antioxidant and anti-inflammatory activities of essential oils: a short review. Molecules15, 9252–9287. 10.3390/molecules15129252

  • CrossRef
  • Google Scholar

• 41

  Morita M. Nakanishi H. Morita H. Mihashi S. Itokawa H. (1996). Structures and spasmolytic activities of derivatives from sesquiterpenes of Alpinia speciosa and Alpinia japonica. Chem. Pharm. Bull.44, 1603–1606. 10.1248/cpb.44.1603

  • CrossRef
  • Google Scholar

• 42

  National Research Council (1996). Guide for the care and use of laboratory animals. Washington, DC: National Academy Press, 1–7.

  • Google Scholar

• 43

  Nawarth H. (1981). Action potential, membrane currents and force of contraction in cat ventricular heart muscle treated with papaverine. J. Pharmacol. Exp. Ther.218, 544–549. 10.1016/s0022-3565(25)32705-9

  • CrossRef
  • Google Scholar

• 44

  Pasricha P. J. (2006). “Treatment of disorders of bowel motility and water flux; antiemetics; agents used in biliary and pancreatic disease,” in Goodman and gilman’s the pharmacological basis of therapeutics. 11th ed. (New York: McGraw-Hill), 983–1008.

  • Google Scholar

• 45

  Pavlović I. Omar E. Drobac M. Radenković M. Branković S. Kovačević N. (2017). Chemical composition and spasmolytic activity of Cymbopogon schoenanthus (L.) spreng. (poaceae) essential oil from Sudan. Arch. Biol. Sci. (Beogr).69, 409–415. 10.2298/ABS160506113P

  • CrossRef
  • Google Scholar

• 46

  Pietrusko R. G. (1979). Drug therapy reviews: pharmacotherapy of diarrhea. Am. J. Hosp. Pharm.36, 757–767. 10.1093/ajhp/36.6.757

  • CrossRef
  • Google Scholar

• 47

  Ponce-Monter H. Campos M. G. Pérez S. Pérez C. Zavala M. Macías A. et al (2008). Chemical composition and antispasmodic effect of Casimiroa pringlei essential oil on rat uterus. Fitoterapia79, 446–450. 10.1016/j.fitote.2008.04.005

  • CrossRef
  • Google Scholar

• 48

  Rehman N. U. Ansari M. N. Ahmad W. Ali A. (2024). Calcium channel inhibitory effect of marjoram (Origanum majorana L.): its medicinal use in diarrhea and gut hyperactivity. Front. Biosci.29, 47. 10.31083/j.fbl2902047

  • CrossRef
  • Google Scholar

• 49

  Rehman N. U. Ansari M. N. Hailea T. Karim A. Abujheisha K. Y. Ahamad S. R. et al (2021). Possible tracheal relaxant and antimicrobial effects of the essential oil of Ethiopian thyme specie (Thymus serrulatus hoschst. Ex benth.): a multiple mechanistic approach. Front. Pharmacol.12, 615228. 10.3389/fphar.2021.615228

  • CrossRef
  • Google Scholar

• 50

  Rehman N. U. Mehmood M. H. Alkharfy K. M. Gilani A. H. (2012). Studies on antidiarrheal and antispasmodic activities of Lepidium sativum crude extract in rats. Phytother. Res.26, 136–141. 10.1002/ptr.3642

  • CrossRef
  • Google Scholar

• 51

  Selim S. A. (2011). Chemical composition, antioxidant and antimicrobial activity of the essential oil and methanol extract of the Egyptian lemongrass Cymbopogon proximus stapf. Grasas Aceites62, 55–61. 10.3989/gya.033810

  • CrossRef
  • Google Scholar

• 52

  Tangpu V. Yadav A. K. (2006). Antidiarrhoeal activity of Cymbopogon citratus and its main constituent, citral. Pharmacologyonline2, 290–298.

  • Google Scholar

• 53

  Van den Brink F. G. (1973). The model of functional interaction. I. Development and first check of a new model of functional synergism and antagonism. Eur. J. Pharmacol.22, 270–278. 10.1016/0014-2999(73)90026-5

  • CrossRef
  • Google Scholar

• 54

  Warrag N. M. Tag Eldin I. M. Ahmed E. M. (2014). Effect of Cymbopogon proximus (mahareb) on ethylene glycol-induced nephrolithiasis in rats. Afr. J. Pharm. Pharmacol.8, 443–450. 10.5897/AJPP2014.4053

  • CrossRef
  • Google Scholar

• 55

  Zhao J. Fan Y. Cheng Z. Kennelly E. J. Long C. (2024). Ethnobotanical uses, phytochemistry and bioactivities of cymbopogon plants: a review. J. Ethnopharmacol.330, 118181. 10.1016/j.jep.2024.118181

  • CrossRef
  • Google Scholar