Abstract

This study aimed at investigating the anti-inflammatory potential of essential oil from rhizome and leaf of Alpinia calcarata Rosc. (ACEO) with the focus of its topical anti-inflammatory activity along with its dominant compounds 1,8-cineole and α-terpineol using mouse ear edema model. ACEOs were analyzed by GC-MS. The anti-inflammatory activity was determined by studying the inhibition of overproduction of proinflammatory mediators—nitric oxide, reactive oxygen species, prostaglandins, cyclooxygenases, and cytokines induced by lipopolysaccharides in murine macrophages. Topical anti-inflammatory and antinociceptive activity was studied by 12-O-tetradecanoylphorbol-13-acetate (TPA) induced skin inflammation and formalin-induced pain model in mice, respectively. Rhizome oil has 1,8-cineole (31.08%), α-terpineol (10.31%), and fenchyl acetate (10.73%) as major compounds whereas the ACEO from leaves has 1,8-cineole (38.45%), a-terpineol (11.62%), and camphor (10%). ACEOs reduced the production of inflammatory mediators in vitro in a concentration-dependent manner. Further, ACEO and its major compounds reduced ear thickness, weight, myeloperoxidase, and cytokines significantly (p < 0.01) in mouse ear. Dose-dependent reduction in flinching and licking in both the phases of pain sensation concludes the topical analgesic effect. Our findings suggest the potency of topical use of ACEOs for inflammatory disease conditions.

Highlights

  • Inflammation is a defending process exhibited by organisms against noxious stimuli, marked by an intensified blood influx to the affected tissue resulting in pain, heat, swelling, redness, and loss of function of the affected part [1, 2]

  • Evidences suggest that the longterm use of nonsteroidal anti-inflammatory drugs (NSAIDs) may induce gastrointestinal ulcers, bleeding, and renal disorders leading to the exploration of less or noninvasive therapeutics which could aid in the treatment strategy of the acute and chronic inflammatory diseases [14]

  • Superoxide anion scavenging activity was measured in sodium phosphate buffer (100 mM, pH 7.4) containing NBT solution (150 μM), NADH solution (468 μM), and different concentrations (1.56–50 μg/mL) of Alpinia calcarata essential oils RO (ACEOs). e reaction was started with the addition of PMS (60 μM) to the mixture followed by incubation at 25°C for 5 min and measurement of the absorbance at 560 nm

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Summary

Introduction

Inflammation is a defending process exhibited by organisms against noxious stimuli, marked by an intensified blood influx to the affected tissue resulting in pain, heat, swelling, redness, and loss of function of the affected part [1, 2]. The main constituents 1,8-cineole (CIN) and α-terpineol (TPN) have been known to act as anti-inflammatory agents in vivo [27, 28]; its topical anti-inflammatory effect and mechanism of action for skin diseases such as atopic dermatitis were never reported Taking this into account, we postulated that ACEO which is rich in monoterpenes like 1,8-cineole and α-terpineol could be effective in preventing TPA-induced acute skin inflammation in mice and inhibit inflammatory mediators in vitro. With the existing knowledge from literatures, this is the first study that reports the detailed profile of volatile constituents, topical anti-inflammatory activity, and in vitro mechanism of action of AC To test this possibility, we have studied the effects of ACEO and main constituents on the TPA-induced cutaneous inflammation. The cytotoxicity of the biologically active ACEOs was evaluated on macrophages, intestinal epithelial cells, human hepatocytes, and keratinocytes in order to assess the effect if targeted for oral or topical application and further gauge the therapeutic edge over the existing drugs

Materials and Methods
In Vitro Anti-Inflammatory Assays
Topical Anti-Inflammatory Activity
Results and Discussion

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