An Overview of Synergistic Effect of Anti-Inflammatory Activity Shown by Moringa Oleifera and Cynodon Dactylon

Abstract

Although inflammation is a protective mechanism, its persistence may lead to chronic disease such as arthritis and diabetes. Conventional anti-inflammatory drugs, though effective, possess serious adverse effects and thus create a demand for safer herbal alternatives. Moringa oleifera and Cynodondactylon are traditionally used medicine plants recognized for their potential antioxidant and anti-inflammatory activities. Their combination resulted in a synergistic effect owing to the diverse phytochemical that include flavonoids, phenolic acids, isothiocyanates, and triterpenoids. The phytochemicals modulate multiple inflammatory pathways by inhibiting COX, LOX, and iNOS enzymes and suppressing cytokines TNF-? and IL-1? via NF-kB regulation. This synergy increases efficacy and safety, indicating that Moringa-Durva can be a promising, natural alternatives in the management of inflammatory disorders.

Keywords

Introduction

Inflammation is a physiological defence mechanism, which the immune system mounts in response to infection, trauma or tissue injury. It is characterised by classical symptoms such as redness, swelling, pain and loss of function. While acute inflammation is protective and aids in tissue repair, chronic inflammation becomes pathological, contributing to the onset of diseases such as rheumatoid arthritis, atherosclerosis, asthma, diabetes, neurodegenerative disorders.

The inflammation process involves a cascade of cellular and molecular events: the activation of immune cells (macrophages, neutrophils), production of reactive oxygen species (ROS) and release of pro-inflammatory mediators like prostaglandins, leukotrienes and cytokines, These mediators are under the regulation of important enzymes, namely cyclooxygenase (COX), lipoxygenase (LOX) and inducible nitric oxide synthase (iNOS).

The conventional therapies mainly involve the use of NSAIDs and corticosteroids, which impede prostaglandins synthesis and immune activation. However, their continuous administration leads to adverse effects such as gastrointestinal ulcers,nephrotoxicity and systemic immunosuppression (Vane and Botting,1998).This limitation has been driving the scientific world towards finding phytochemicals or herbal extracts with anti-inflammatory potential through the use of multiple pathways with minimum side effects,

Moringa Oleifera and cynodondactylon  are important medicinal plants, well  traditional systems of medicine such as Ayurveda and Siddha. Both  possessed a wide range of pharmacological properties including anti-inflammatory, Antioxidants, Antimicrobial and wound healing effects. Recent studies have indicated that there combine use provide enhanced anti-inflammatory activity, compared to individual  extracts, which may be due to the synergistic interaction of bioactive compounds.

Synergy, in herbal pharmacology, represents the combined effect of two or more components exhibiting a total effect that is greater than the sum of each individual components. For example, Moringa Oleifera and Cynodon Dactylon exihibit Synergy in their anti-inflammatory activity by simultaneously inhibiting Inflammatory enzymes,  suppressing the production of cytokines, enhancing antioxidant defences. This paperreviews the phytochemistry, mechanisms of the synergistic combination, and  experimental validation, thereby  presenting it as a potential natural therapeutic agents for inflammatory disorders.

1.Moringa Oleifera

Moringa oleifera Lam. (M. oleifera) is widely distributed and utilized in tropical and sub-tropical regions of the world, and is mainly native to India and Africa. It is often referred to as drumstick tree or horseradish tree, and also called “miracle tree”, or “natural gift”, or “mother’s best friend”, due to the high nutrients in leaves with protein, minerals, and β-carotene . The M. oleifera leaves can be eaten in multiple ways, and can maintain nutrient levels for a long time when stored as dried powder. Therefore, some regional and international relief organizations are putting a great deal of emphasis on developing the leaves of M. oleifera as a nutritional supplement in some Africa countries.

Traditionally, in some countries such as India , Pakistan and Uganda, M.oleifera has long been in wide use to diabetes, obesity, hysteria ,scurvy and even tumours . It is reported that M.oleifera contains many phytoconstituents such as flavonoids  ,alkaloids , saponins, saccharides, glucosinolates, tannis , phenolic acids , and nitrile glycoside, etc.

All those complicated natural phytochemistry contribute to its numerous pharmacological activities.

For eg- the leaves of M.oleifera usually display good anti-inflammatory  , anti-cancer , anti-oxidant , anti-bacterial , hepatoprotective , cardioprotective , anti hypertension, hypolipidemic, hypoglycemic , activities and so in the : seed display distinct antimicrobial , antidiabetic  , anti inflammatory  activities, etc. and the roots can also possess some biological activities including anti-inflammatory , anti-cancer, anti- ulcer, antifertility , anti-urolithiatic activity, and so on therefore, M. Oleifera has been widely studied in recent years due to its enormous potential as a source of healthy food with medicine values.

2.Cynodon Dactylon

Cynodondactylon (Poaceae), a hardy perennial grass, is one of the most commonly occurring weeds in India. In Hindi it is known as dhub, doob or harialil; other common names include durba (Bengali), garikoihallu (Kanarese), durua (Marathi), durua or haritali (Sanskrit), arugampullu (Tamil), garikagoddi (Telugu) and dhubkhabbal (Punjabi) (Sastry and Kavathekar, 1990). Cynodon has a renowned position in Indian systems of medicine and many parts of the plant are assumed to have medicinal properties. Doob ghas is a valuable herbal medicine and used as first aid for minor injuries (Oudhia, 1999a, b). Farmers traditionally apply crushed leaves to minor wounds as a styptik to stop bleeding (Oudhia, 2001). The whole plant is extremely beneficial externally in wounds and the paste of the plant is applied on forehead in headache (Paranjpe, 2001). The roots in the form of paste with water are taken internally against fevers (Natarajan and Paulsen, 2000). The aqueous fluid extract of the rhizome is used as anti-inflammatory, diuretic,anti-emetic, purifying agent and also in dysentery (Ahmed et al., 1994; Kirtikar and Basu, 1980).

Cynodon plant is useful for pains, inflammations and toothache ,various scientific studies have been carried out on Cynodondactylon and various pharmacological activities have been reported. It has been reported to possess anti diabetic , cardio protective and immunomodulatory activities. Since no detailed scientific data is available regarding the anti inflammatory activity of cynodondactylon,the present study was designed to explore the same.

Botanical Description

Botanical Comparison of Moringa Oleifera and Cynodon Dactylon

Phytochemical Composition

Experimental animals: Male albino wistar rats weighing between 200-250 g and albino mice (25-30 g) were used. Institutional Animal Ethics Committee approved the experimental protocol. Animals were maintained under standard conditions in an animal house approved by Committee for the Purpose of Control and Supervision on Experiments on Animals (CPCSEA).

• Plant material: The leaves of Moringa oleifera Lam were collected from Manipal. Udupi, INDIA, and it was authenticated by Dr. K. Gopalakrishna Bhat, Professor of Botany (Rtd.), Taxonomy Research, Centre, Department of Botany, Poornaprajna College Udupi-576 101, Karnataka.
• Chemicals: Carbopol 940, Diclofenac gel, Propylene glycol 400, Ethanol, Methyl paraben, Propyl paraben, Ethylenediaminetetraacetic acid (EDTA). Triethanolamine, Carrageenan were purchased from the market
• Plant extract preparation Leaves were shade dried and coarsely powdered with cutter mill. The powder was subjected for extraction using Soxhlet apparatus and ethanol as solvent. The extracts were concentrated on a water bath at a temperature below 50° C, and dried in a desiccator, stored in refrigerator until used.
• Animals: Adult, female, Wistar rats weighing between 150-200 g were obtained from the Central animal house and maintained under constant conditions (temperature 25+2°C, humidity 40-60%, 12 h light and 12 h dark cycle). During this time of maintenance, they received a standard diet and water ad libitum. The experiment was approved by the Institutional animal ethics committee (IAEC/KMC/29/2015). Preparation of topical gel: The extract gels were prepared using dried alcoholic extract of Moringa oleifera Lam leaves. The water required for these formulation divided into two parts. In one part, the exact amount of extract was dissolved to which calculated amount of propylene glycol 400 & ethanol was added while in other, carbopol-940 was dissolved to which methyl paraben, propyl paraben, EDTA were added. Both of these solutions were mixed in a beaker to which triethanolamine was added to obtain a gel consistency and pH. Blank gel was prepared in a similar fashion, Methodology: The Wistar albino. into four groups of six rat in each. The group-1 animals had received topical gel without sout containing ethanolic leaves extract (blank gel), while group-3 and 4 animals had received ethanolic leaves extract were divided gel at dose of 2% and 4% respectively. The group-2 had received standard diclofenac 0.5% topically 10 left leg one hour before the injection of carrageenan, Assessment of anti-inflammatory activity: The anti-inflammatory activity of topical Moringa oleifera Lam, against acute inflammation was tested using carrageenan induced rat paw edema model. Pedal inflammation was produced according to the method described by winter et al. The edema was induced by injecting 0.1ml of carrageenan (1% w/v) in normal saline into the sub-plantar region of left hind paw The paw was marked with ink at the level of lateral malleolus. The paw volume measured by Plethysmometer at 0, 30, 60, 120, 180, and 240 minutes after carrageenan injection.The percentage inhibition of edema was calculated using the formula,

Percentage inhibition of edema(1-Vt/Vc) x100

Where, Vt mean volume of paw edema in drug treated group Ve mean volume of paw edema in the control group

• Statistical analysis: The data was analysed using one-way ANOVA followed by post hoc test. P value less than 0.05 was considered significant, of the rats subjected to 2% gel gel leaves extract of Moringa oleifera Lam, was found to be significantly reduced when compared with control at 4th hour of treatment, P=0.04 (Table-2), the percentage reduction in paw volume at 4th hour was 21% and was comparable to the standard (0.5% Diclofenac, showed 26% reduction). Inflammation is the protective response that involves immune cells, blood vessels & molecular mediators. The paw edema in rats, induced by carrageenan locally is inhibited by anti-inflammatory agents given by different routes having their own advantages. Carrageenan induces biphasic inflammation-early phase inflammation (due to histamine, serotonin, bradykinin) and late phase inflammation (due to prostaglandins) The late phase is inhibited by NSAIDs (Diclofenac) but not early phase7.10.12. The topical gel of leaves extract of Moringa oleifera Lam. having 2% strength showed reduction in paw volume at 240 minute reflecting the probable mechanisms of cyclooxygenase enzyme inhibition. and thereby causing a production of prostaglandins. The earlier literature reported ethanolic extract of leaves extract of Moringa oleifera Lam given orally had significant reduction of paw volume at 3 hr13.Pharmacological basis of synergy When moringa Oleifera and cynodondactylon are combined, there phytoconstituents exert a simultaneous action on multiple molecular targets, leading to a synergistic pharmacodynamic effect.

(a)  Multi-target Enzyme Inhibition

The active flavonoids of moringa inhibit COX-2 and lipoxygenase, thus reducing prostaglandin and leukotriene synthesis.

Durva triterpenoids and flavones exert an additional inhibitory effect on COX and phospholipase A2, thereby reducing arachidonic acid metabolism.

Result: Stronger inhibition of pro-inflammatory mediators and reduced tissue edema.

(b) Modulation of Cytokine and Gene Expression

Moringa suppresses TNF-α, IL-1β and IL-6 production.

Durva downregulates IL-6, IL-8 and MCP-1, avoiding neutrophil infiltration.

Result: Broader suppression of inflammatory signaling and immune cell activation.

(c) Antioxidant Synergy

Both plants are rich in flavonoids and polyphenols, which scavenge free radicals such as superoxide anion, hydroxyl radicals, and nitric oxide. This reduces oxidative stress, which is one of the prime triggers for chronic inflammation.

(d) NF-kB pathway inhibition

The NF-kB transcription factor controls genes responsible for cytokine production.

The isothiocyanates in Moringa and the apigenin in Durva synergistically inhibit NF-kB activation, blocking inflammatory gene expression. This allows for long-term control of inflammatory processes.

(e) Stabilization of cell and lysosomal membranes

Both herbs stabilize lysosomal membranes, preventing leakage of proteolytic enzymes that damage nearby tissues. This reduces cellular injury and edema formation.

5. Evidence From Experiment

A. Research into Moringa oleifera

Carrageenan-induced paw edemamodel: Ethanolic leaf extract at 300 mg/kg reduced paw-edema by 60-70% comparable to indomethacin.

Formalin-induced arthritis model: Moringa significantly inhibited joint inflammation and pain.

In vitro: Extract inhibited the gene expression of COX-2 and iNOS, confirming anti-inflammatory potential.

B. Studies on Cynodondactylon

Acetic acid-induced writhing test (mice): Ethanolic extract demonstrated good analgesic and anti-inflammatory activities.

Formalin-induced paw edema-: significantly reduced inflammatory swelling and release of cytokines.

Protein denaturation assay: It demonstrated considerable inhibition of heat-induced protein denaturation, reflecting membrane stabilization.

C. Combined Extract (Synergistic Study)

The nitric oxide and prostaglandin synthesis were significantly inhibited in the hydro-alcoholic mixture of Moringa-Durva in a 1:1 ratio compared to its individual extracts.

In vitro macrophage model:The combined extract significantly deceased TNF-αAnd IL-6 production, indicating a synergistic anti-inflammatory action. In vivo: The combination reduced the volume of the paw by >75%, while the individual extracts achieved 50-60%6.Prospects for formulations and application

A. Formulation type

1. Topical gel or cream: For arthritis joint pain , and local inflammation

2. Oral capsule or suspension : for systemic inflammatory conditions

3. Herbal hydrogel patch for sustained drug delivery and a localised anti-inflammatory effect.

4. Nanoemulsion or liposomal formulations: To improve the bioavailability of flavonoids.

B. Ideal Ratio

Experimental data indicate that a 1:1 or 2:1 ratio (Moringa:Durva) gives optimum synergistic activity, which can be optimised based on extract standardization.

C. Evaluation Parameters

In vitro: Proteins denaturation assay, albumin stabilization assay, No inhibition assays. In-vivo: carrageenan-induces edema;fomalin arthritis; granuloma pouch model.Biochemicalmarkers : TNG-α, IL-1β, COX-2, PGE2, CRF .

7. Advantages of the Combined Extract

Benefit

Explanation

1. Synergistic efficacy

Several bioactive compounds act on various inflammatory targets.

Reduce side effects

Lower individual dose requirement because of greater efficacy

2. Anti-oxidant protection

Prevents oxidative stress- related inflammation

3. Biocompatibility

Natural, non- toxic, and eco - friendly

Dual action

Both anti- inflammatory and analgesic

Potential for chronic useSuitable for long-term inflammatory disorders

8. Possible therapeutic uses

• Rheumatoid arthritis and osteoarthritis
• Muscular and joint inflammation
• Skin inflammation ( eczema)
• Gastrointestinal inflammation disease
• Wound healing and tissue regeneration
• Anti-allergic and asthma supportive therapy

9. Safety and toxicity

Both moringa and Cynodondactylon are non-toxic at their therapeutic dosage. Acute oral toxicity studies show no mortality or behavioural changes up to 2000 mg/ kg . Their nutritional composition of proteins, vitamins, and minerals adds to their safety profile .

CONCLUSION:

The present review establishes that Moringa oleifera and Cynodondactylon possess remarkable anti-inflammatory potential, both as single entities and in combination. Their synergistic effect arises from the diverse phytoconstituents-falvanoids, phenolic acids, terpenoids, alkaloids and isothiocynates which act through multiple biochemical pathways such as inhibition of COX, LOX and iNOS enzymes, suppression of cytokines (TNF-α,IL-1β, IL-6), and modulation of the NF-kB signalling cascade. The combined use of the plants increase efficacy and reduces the required dosage, along with minimizing toxicity compared to synthetic drugs.

 The synergistic pharmacological interaction is confirmed by experimental evidence showing that the Moringa-Durva combination produces more effective inhibition of inflammation, oxidative stress and tissue edema. Further, their safety profile and biocompatibility make them ideal candidates for long-term therapeutics use in managing chronic inflammatory disease like arthritis, asthma and gastrointestinal disorders.

 Hence, the combination of Moringa oleifera and Cynodondactylon stands out as a promising alternative in nature to conventional anti-inflammatory drugs, which could be given as topical gels, oral preparations and new formulations such as hydrogels or nanoemulsions for better bioavailability and patient compliance.

ACKNOWLEGEMENT:

An overview of Extraction and Synergistic effect of Anti-Inflammatory activity shown by Moringa Oleifera and Cynodon Dactylon. Her insightful suggestions and expert supervision have been instrumental in shaping the quality and depth of this work. The author also extends their appreciations to their institution and colleagues for providing the necessary resources and academic environment that facilitated this study.

Conflict of interest:

The author declared no conflict of interest with respect to the authorship and publication of this article.

 Availability of data and material:

 All the data used in our article are available from publicly accessible sources such as PubMed, Elsevier, Wikipedia.

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