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Methanolic extract of Ephedra ciliata promotes wound healing and arrests inflammatory cascade in vivo through downregulation of TNF-α

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Abstract

Chronic wounds may lead to the development of various pathological conditions such as diabetic foot ulcers and pressure sores. The current study evaluated wound healing and anti-inflammatory potentials of methanolic extract of Ephedra ciliata using series of in vivo models. Methanolic extract of Ephedra ciliata was prepared by maceration (Ec.Me). Qualitative and quantitative (HPLC) phytochemical and metal analyses were conducted to explore the chemical and metal profiles of Ec.Me. Safety profile (behavioural) and, antimicrobial, antioxidant, wound healing, anti-inflammatory and anti-angiogenic potentials of Ec.Me were evaluated using well-established in vitro and in vivo models. ELISA assay was performed to estimate the effects of Ec.Me treatment on serum levels of TNF-α. HPLC analysis identified quercetin as one of the major compounds in Ec.Me. Safety study data showed that Ec.Me was safe up to the dose of 2000 mg/kg. Antimicrobial assay data showed that Ec.Me was active against bacterial (Staphylococcus aureus) as well as fungal (Candida albicans and Aspergillus niger) strains. Ec.Me showed modertate antioxidant potential in in vitro and in vivo models. Data of excision and burn wound healing models showed that Ec.Me, promoted wound closure in a dose and time-dependent manner. Treatment with 20% Ec.Me cream and heparin showed almost the same effects with no statistical differences (p > 0.05). Ec.Me also showed time-dependent anti-inflammatory activities in both acute and chronic models. In carrageenan model, treatment with 200 mg/kg of Ec.Me showed comparable anti-inflammatory effects (p > 0.05) with quercetin and indomethacin throughout the study. In cotton pellet granuloma model treatment with 200 mg/kg of Ec.Me and indomethacin inhibited granuloma formation significantly better (p < 0.05) as compared with the rest of the treatment groups. Histopathological examination of skin samples showed marked improvement in architecture with minimal infiltration of inflammatory cells. Data of in vivo angiogenesis assay showed marked improvement in vessels length, density, branching points, total segments and total nets after treatment with Ec.Me, indicating no toxic effects towards vasculature development. Significant (p < 0.05) downregulation of TNF-α was observed in serum samples of animals treated with Ec.Me. Based on data of the current study, it is concluded that quercetin-rich extract of Ephedra ciliata has wound healing and anti-inflammatory potentials via downregulation of TNF-α. Moreover, it is suggested that the antimicrobial activity of Ec.Me prevented microbial invasion, thus promoted natural wound healing mechanisms as well.

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Acknowledgements

We would like to acknowledge the Faculty of Pharmaceutical Sciences, Government College University Faisalabad, for providing the facilities to conduct this study. This project was financially supported by the Higher Education Commission of Pakistan Research Grant (No: 21-8128/SRGP/R&D/HEC/2018; Muhammad Asif [PI]). We are grateful to Dr. Imran, Associate Professor, Department of Pharmacy, Bahauddin Zakariya University (BZU), Multan and Dr. Liaqat Hussain, GC University Faisalabad for generous donation of quercetin.

Funding

This project was supported by Higher Education Commission of Pakistan Research Grant (No: 21-8128/SRGP/R&D/HEC/2018).

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    Authors and Affiliations

    1. Faculty of Pharmaceutical Sciences, Government College University of Faisalabad, Faisalabad, Pakistan

      Hafiza Sidra Yaseen, Muhammad Asif, Malik Saadullah,  Mahrukh, Sajid Asghar, Rizwan Rashid Bazmi & Hafiz Muhammad Yousaf

    2. Department of Pharmacy, The Islamia University of Bahawalpur, Bahawalpur, Pakistan

      Muhammad Asif

    3. FMH College of Medicine and Dentistry, Lahore, Pakistan

      Muhammad Usman Shams

    4. Department of Pharmacology, Punjab University College of Pharmacy, University of the Punjab, Lahore, Pakistan

      Mohammad Saleem

    5. Sheikh Zayed Hospital, Lahore, Pakistan

      Maria Yaseen

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    1. Hafiza Sidra Yaseen
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    Authors declare no conflict of interest in the current work. Present work was conducted as part of M.Phil thesis by Hafiza Sidra Yaseen under the supervision of Muhammad Asif (PI) and Malik Saadullah (Co-PI). Hafiza Sidra Yaseen, Muhammad Asif, Malik Saadullah, Mohammad Saleem and Mahrukh contributed equally in this study.

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    Yaseen, H.S., Asif, M., Saadullah, M. et al. Methanolic extract of Ephedra ciliata promotes wound healing and arrests inflammatory cascade in vivo through downregulation of TNF-α. Inflammopharmacol 28, 1691–1704 (2020). https://doi.org/10.1007/s10787-020-00713-7

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