In-Vitro Evaluation of Antispasmodic Activity of Rhizomes of Curcuma Amada Roxb.

 Dr. Sundhararajan R, Sanjukta Sainath Singh*, Jenifer J, Keerthana P, Kiruthika

Mohammad Sathak A.J. College of Pharmacy, Sholinganallur, Chennai, Tamil Nadu.

*Correspondence: sanjuktagharami@gmail.com; Tel.: 9940406560

DOI: https://doi.org/10.71431/IJRPAS.2025.4308

INTRODUCTION

Food ingestion and digestion, nutritional absorption, and waste product removal are all handled by the gastrointestinal tract. The GIT is composed of the mouth, esophagus, stomach, small and large intestines, and anus. Food ingestion and digestion, nutritional absorption, and waste product removal are all handled by the gastrointestinal tract. The GIT is composed of the mouth esophagus, stomach, small and large intestines, and anus.[1] The digestive system includes the pancreas, liver, gall bladder, and gastrointestinal tract. It is a network of blood vessels that provides blood to the organs and transports nutrients from other bodily parts. Furthermore, hormones and nerves work together to regulate the digestive systems’ performance. The bacteria that live in our GI tract are essential to our immune system,

digestion, and overall health. GIT disorders are any illnesses that impact the gastrointestinal tract. This group includes any symptoms that show up in the middle or lower part of the digestive tract. [2].

Herbal treatments have been utilized for ages to cure a wide range of disorders by traditional medical systems and ethnic healers. It is estimated that 80% of people nowadays treat a range of illnesses with traditional herbal treatments. This is partly due to its availability and absence of adverse effects. Traditionally, a range of herbs and medications have been used to treat digestive system disorders. Different plant parts, such as leaves, roots, rhizomes, etc., are used in the treatment. These have more potency and are less toxic than modern medicines[3]. Numerous plants and herbs that have been used traditionally to treat and prevent GIT problems have undergone scientific investigation. These plants and herbs have been shown to have bioactive substances including flavonoids and tannins that have therapeutic effects on the human body. Traditionally, plants like Zingiber officinale[4], Glycyrrhizauralensis[5], Artemisia vulgaris[6], and others have been utilized to treat GIT issues. Curcuma amada is well known as mango ginger, it belongs to the genus Curcuma and family Zingiberaceae. The GCMS analysis revealed the presence of 83 components, of which β-Myrcene (6.6 %), epicurzeronone (2.5 %), squalene (2.2%), α-acaridial (1.8 %), β-pinene (1.8 %), 2,6, 11,15-Tetramethyl-hexadeca2,6,8,10,14-pentaene (1.7 %) and aromadandrene (1.1 %) were the major components reported.[7] The rhizome is used in the treatment of antimicrobial[8] , antifungal activity[9], anticancer activity[10], antibacterial agent[11], Mango ginger is widely regarded in Ayurvedic and Unani medicinal systems as a  digestive aid, aphrodisiac, antipyretic, emollient, diuretic, laxative, and expectorant as well as a cure for  biliousness, itching, skin disorders, bronchitis, asthma, hiccups, and    inflammation caused by accidents[12], antidiabetic activity[13], antihyperglycemic effects[14], anti-inflammatory activity.[15-17]. Therefore, the purpose of this study was to investigate the antispasmodic effects of Curcuma amada rhizomes on the in vitro voluntary motility and contractility of the smooth muscle of the chicken ileum. Using chicken ileum, the study assesses the antispasmodic effect of Curcuma amada rhizomes on intestinal contraction.

MATERIALS AND METHODS

Rhizome Collection and Authentication:

A fresh Curcuma amada Roxb (Mango ginger) rhizome was collected from the local market of Chennai, Tamil Nadu in Jan 2025 (1157.05022508/PCOG002-ACF). The identification and authentication of the plant was done by Dr. K.N. Sunil Kumar, Research Officer/Sci-II and HOD, department of pharmacognosy, Siddha central Research Institute, Central council for Research in Siddha, Ministry of Ayush, Arumbakkam, Chennai, India. The rhizome skin was removed and cut into small pieces. Then the pieces were washed by using distilled water, shade dried, and mechanically grinded. The coarse powder (100 gms) was homogenized and pulverized by taking mesh size 40 and stored in an air-tight container free from moisture and contamination.

Chemicals and glassware:

The glassware used for this  study were test tubes, a round-bottom flask with stopper, a heating mantle, a soxhlet apparatus, Sherrington’s rotating drum, filter paper, a curve needle, thread, and syringes (2 ml) procured from Southern India Scientific Corporation, R.A.Puram, Chennai, Tamilnadu, India. All chemicals used for this study were obtained from Sudhkar biological and chemicals private limited, Vepery high road, Chennai. Chicken ileum was freshly collected from local vendor and kept in ice cold oxygen tyrode solution with aeration.

Preparation of Histamine induced contraction of chicken ileum:

Chicken ileum was suspended in bath containing Tyrode solution (Composition mM: NaCl, 136.7; KCl 2.68; CaCl2, 1.8; NaHCO3,11.90; NaH2PO4, 0.42; MgCl2, 1.05; glucose, 5.55)

maintained at 37±0.5°C. A stream of air was bubbled through the organ tube (1 bubble/sec). One end of the ileum was attached to an S-shaped aerator and the other was attached to an isotonic

frontal writing lever to the drum. The tissue was allowed to equilibrate for 45 min under a load of 500

mg. Contact time of 60 sec, and baseline of 30 sec time cycle opted for proper recording. Cumulative Dose Response Curve (DRC) was recorded on a kymograph for histamine (1mg/ml) in the absence and the presence of the aqueous extract of Curcuma amada Roxb in different concentrations such as 50 mg/ml, 75 mg/ml, and 100 mg/ml on kymograph with the use of Sherrington’s recording drum. The same procedure was carried out for DRC of histamine in the presence of Chlorpheniramine maleate (CPM) as a standard drug. The percentage inhibition of extract and the standard drug was calculated and the Graph was plotted by taking the log dose versus the height of the response curve. Percentage inhibition of extract and standard drug was calculated[18,19].

Preparation of Acetylcholine induced contraction of chicken ileum:

The same procedure was followed up to baseline of 30 sec time cycle opted for proper recording for the above preparation. Cumulative Dose Response Curve (DRC) was recorded on kymograph for acetylcholine (1mg/ml) in the absence and the presence of the aqueous extract of Curcuma amada Roxb in different concentrations such as 50 mg/ml, 75 mg/ml, and 100 mg/ml on kymograph with the use of Sherrington’s recording drum. The same procedure was carried out for DRC of acetylcholine in the presence of atropine (0.6 mg/ml) as a standard drug. The percentage inhibition of extract and standard drug was calculated. A graph was plotted by taking log dose versus height of the response curve. Percentage inhibition of extract and standard drug was calculated.[20,21]

RESULT

The effect of the aqueous extract of Curcuma amada Roxb (AECAR) on chicken ileum preparation was carried out using histamine, and acetycholine-induced contraction. Atropine and chlorpheniramine maleate were used as antagonists to acetylcholine and histamine, respectively. Both acetylcholine as well as histamine have shown dose-dependent activity in combination with atropine and chlorpheniramine maleate respectively on chicken ileum with DRC of highest concentration of 1.6 ml with DRC of 7.0 cm, 12.0 cm, 7.4 cm, 13.9 cm. In the same way, AECAR 50 mg/ml, 75mg/ml, and 100 mg/ml showed dose-dependent activity at the maximum concentration of 1.6ml but 100mg/ml showed potent activity when compared with acetylcholine and histamine with DRC of 16.0 cm,12.0 cm, 16.5 cm, 13.9 cm respectively. Other doses of 50 mg/ml and 75 mg/ml of AECAR with different concentrations also have resulted in significant activity as shown in Table 1 and 2. The graphical representation was shown in Figures 1 and 2.

Table 1: DRC of Aqueous extract of Curcuma amada Roxb using histamine (agonist) and Chlorpheniramine Maleate (antagonist)

Table 2: DRC of Aqueous extract of Curcuma amada Roxb using Acetylcholine (agonist) and atropine (antagonist)

Figure 1: Graphical representation for the DRC of AECAR using Histamine and Chlorpheniramine maleate.

Figure 2: Graphical representation for the DRC of AECAR using Acetylcholine and Atropine.

DISCUSSION

The phytochemical screening study was reported. It showed the presence of alkaloids; it acts as anticholinergics. They block the action of acetylcholine, a neurotransmitter that stimulates muscle contractions, by binding to muscarinic receptors in smooth muscles. This inhibition leads to muscle relaxation and reduces spasm. Alkaloids can block calcium channels in smooth muscle cells. Calcium ions play a crucial role in muscle contraction. The influx of calcium inhibition of alkaloids, saponins, terpenoids prevents muscle contractions and reduces spasms. Saponins can interact with cell membranes and modulate ion channels, which help in regulating muscle contractions. This interaction can lead to the relaxation of smooth muscles and reduction of spasms. It possesses anti-inflammatory properties that help in reducing inflammation in smooth muscles. By decreasing inflammation, saponins can alleviate the spasms associated with muscle irritation or injury. Terpenoids can interact with various receptors in the body, such as TRPM8 (transient receptor potential melastatin activates TRPM8 receptors, creating a cooling sensation that helps to relax smooth muscles and reduce spasms. Tannins can cause the contraction of smooth muscle cells by binding to proteins in the muscle tissue. This astringent effect can help reduce muscle contractions and alleviate spasms. Flavonoids are potent antioxidants that can neutralize free radicals and reduce oxidative stress in muscle tissues. Oxidative stress can contribute to muscle spasms, so reducing it helps in relaxing muscles and alleviating spasms. Chicken ileum was used due to the presence of various receptors such as muscarine, serotonin, histaminic, GABAergic, and adrenoreceptors. In many natural plants, they contain some chemical constituents that may have an antispasmodic effect. These natural plants are new, effective, and harmless when compared to synthetic medicines. AECAR with the dose of 50 mg/ml, 75 mg/ml, and 100 mg/ml in different concentrations like 0.1 to 1.6 ml. From this 100mg/ml dose showed significant activity when compared to standards. The reports showed it indicated significant activity in a dose-dependent manner same as standard. ^[22-25]

CONCLUSION

The rhizome of aqueous extract of Curcuma amada Roxb has high significant medicinal value but they have not explored so far hence we have used the rhizome of Curcuma amada Roxb to study antispasmodic activity. The results of the study have shown that AECAR possesses anti-spasmodic activity. Therefore, with the help of this research work exact chemical constituents, mechanisms of action responsible for antispasmodic activity which can be helpful in the treatment of smooth muscle spasms, abdominal cramps, and irritable bowel syndrome, and other pharmacological activities of AECAR can be found in future.

ACKNOWLEDGEMENT

We thanks to management and staffs of Mohamed Sathak AJ College of Pharmacy Chennai, Tamil Nadu, India for providing necessary facilities and full support throughout the research in a successful manner.

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