The essential oil from Citrus limetta Risso peels alleviates skin inflammation: In-vitro and in-vivo study (original) (raw)
Abstract
Ethnopharmacological relevance
Citrus fruit peels are traditionally used in folk medicine for the treatment of skin disorders but it lacks proper pharmacological intervention. Citrus limetta Risso (Rutaceae) is an important commercial fruit crops used by juice processing industries in all continents. Ethnopharmacological validation of an essential oil isolated from its peels may play a key role in converting the fruit waste materials into therapeutic value added products.
Aim of the study
To evaluate the chemical and pharmacological (in-vitro and in-vivo) profile of essential oil isolated from Citrus limetta peels (Clp-EO) against skin inflammation for its ethnopharmacological validation.
Materials and methods
Hydro-distilled essential oil extracted from Citrus limetta peels (Clp-EO) was subjected to gas chromatography (GC) analysis for identification of essential oil constituents and its anti-inflammatory evaluation through in vitro and in vivo models.
Results
Chemical fingerprint of Clp-EO revealed the presence of monoterpene hydrocarbon and limonene is the major component. Pre-treatment of Clp-EO to the macrophages was able to inhibit the production of pro-inflammatory cytokines (TNF-α, IL-6, IL-1β) in LPS-induced inflammation as well as the production of reactive oxygen species (ROS) in H2O2-induced oxidative stress. In in-vivo study, topical application of Clp-EO was also able to reduce the 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced ear thickness, ear weight, lipid peroxidation, pro-inflammatory cytokines production and ameliorate the histological damage in the ear tissue. In-vitro and in-vivo toxicity study indicate that it is safe for topical application on skin.
Conclusion
These findings suggested the preventive potential of Clp-EO for the treatment of inflammation linked skin diseases.
Introduction
Citrus are among the most popular fruit crops in the world. Citrus fruits are an important component of the daily diet in many parts of the world (Liu et al., 2012). Citrus fruits and fruit products are globally important from nutritional and economic perspectives (Rawat et al., 2015). Citrus fruits are traditionally in use for the management of skin ailments (Noh et al., 2015; Apraj and Pandita, 2016) and it was scientifically validated against the skin inflammation (Herath et al., 2016). The compounds (5-Hydroxy-3,6,7,8,3',4'-hexamethoxyflavone, nobiletin) isolated from citrus fruit peels exhibited the anti-inflammatory effect against 12-O-tetradecanoylphorbol-13-acetate (TPA) -induced skin inflammation through the down-regulation of iNOS, COX-2 and NF-kβ (Lai et al., 2007, Choi et al., 2007) in LPS-induced inflammation. Citrus limetta Risso (Rutaceae), commonly known as sweet lime is one of the important commercial Citrus fruit crops used by juice processing industries in the all continents (Anwar et al., 2008). C. limetta comprised of 8–10% peel, which is perishable waste material creates a big challenge in processing industries as well as pollution monitoring agencies (Manthey et al., 2001). Interestingly, C. limetta fruit peels possess various pharmacological activities (Bharti et al., 2014) due to the presence of flavonoids in major quantity (Mohanty et al., 2015). One of the important products of citrus fruit peels is the essential oil, which is pleasant sensory characteristics (Mondello et al., 2005) and it is broadly used in cosmetic and pharmaceutical products (Kondo et al., 2000, Misharina and Samusenki, 2008).
The skin is the largest organ in the body and plays a central role in host immunological defenses. The regulation of skin defense mechanisms is crucial; an inappropriate or misdirected immune activity is implicated in the pathogenesis of variety of acquired inflammatory skin disorders (Kupper and Fuhlbrigge, 2004). Skin diseases linked with inflammation are the most common problem in dermatology which has significant impact on the quality of life of human being and represent an enormous financial trouble (Fuchs et al., 2001). They are in many forms of skin disorders includes occasional rashes accompanied by skin itching, redness and chronic conditions such as dermatitis, rosacea, seborrheic dermatitis, and psoriasis (Maurya et al., 2014, Javed et al., 2013). The secretions of cytokines by keratinocytes in response to injury, particularly tumor necrosis factor alpha (TNF-α) and interleukin (IL-1β), are key mediators of the cutaneous inflammatory response (De Vry et al., 2005). Currently used conventional non-steroidal anti-inflammatory drugs (NSAID) and steroidal anti-inflammatory drugs (SAID) treat acute inflammatory disorders have not been successful to cure chronic inflammatory disorders. Since the critical etiology and exacerbating mechanisms are not completely understood, it is difficult to develop a magic bullet for chronic inflammatory disorders (Kim et al., 2004). Experimental evidences has shown that exposure of skin to 12-O-tetradecanoylphorbol-13-acetate (TPA) induces a pleiotropic tissue response encompassing a strong inflammatory reaction similar to that observed in skin diseases (Kondo et al., 2000). Therefore, this study intended to investigate the chemical composition and in-vitro, in-vivo anti-inflammatory potential of essential oil extracted from the peel of Citrus limetta (Clp-EO) to support its ethnopharmacological claims.
Section snippets
Collection of plant materials and extraction of essential oil
The fruits of Citrus limetta were purchased from the local market of Lucknow India and identified by Dr. Narendra Kumar (Department of Botany and Pharmacognosy, CSIR-CIMAP Lucknow, India). A voucher specimen was deposited in the herbarium of the institute under repository number C052. Fresh fruits were washed, peeled off and 1000
g fresh peels subjected to hydro-distillation in a Clevenger's apparatus for 3
h for extraction of essential oil. The essential oil was measured directly in the
Composition of essential oil isolated from Citrus limetta (Clp-EO)
The extractive yield of isolated Clp-EO was 0.8% (v/w). The GC-MS chromatogram tracing and quantitative analysis of active constituents is shown as Fig. 1. GC-MS analysis demonstrated the presence of 30 compounds. Out of 30, 13 compounds were contributed the 96.8% of total essential oil composition. Monoterpene hydrocarbon contributes a major proportion in Clp-EO and limonene (91.8%) was found as a major component of the oil. The compounds content more than 0.1% in Clp-EO is depicted in Table 2.
Discussion
An exploration of possible pharmacological effects along with characterization of the bioactive compounds present in fruit waste play a key role in converting the fruit waste materials into therapeutic value added products. Citrus fruit peels are waste material which creates a big challenge in fruit processing industries (Manthey and Grohmann, 2001). Citrus-processing industries generate huge amounts of wastes every year, and citrus peel waste alone accounts for almost 50% of the wet fruit
Conclusions
Results of the study demonstrate that topical application of Clp-EO is an effective anti-inflammatory treatment in the TPA-induced skin inflammation. This study suggests the suitability of Clp-EO to be incorporated in skin care formulations for pharmaceutical purposes with special reference to skin inflammation.
Acknowledgments
We acknowledge the Council of Scientific and Industrial Research (CSIR), New Delhi, India for financial support through networking project BSC-0203.
Conflict of interest
The authors declare that there are no conflicts of interest.
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Citrus peel (CP) accounts for about half (40–50%) of the fruit weight. During citrus processing (juicing and canning), thousands of tons of CP solid waste is generated, which is an important source of bioactive components such as phenolic compounds, essential oil (EO), carotenoids and ascorbic acids (Bustamante et al., 2016; Maurya, Mohanty, Pal, Chanotiya, & Bawankule, 2018; Raspo, Vignola, Andreatta, & Juliani, 2020; Singh et al., 2020). Treatment of CP is a major problem in the citrus by-product processing industries, as only a small quantity of it is utilized, while a large amount is buried and burned that can pollute the environment and waste resources.
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The peel, as a considerable part of sweet lemon (8–10%), is generated in the juice processing factories, disposed in the landfills as agricultural waste and created serious environmental problems [15]. While, it is an interesting source of valuable ingredients such as phenolics, flavonoids (in major quantity), essential oils and pectin [16]. Thus, the usage of sweet lemon peels (SLP) in the recovery of value-added products, could decrease environmental pollutions [17].
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