Antioxidant, Antimicrobial, Antidiabetic, Antiglycation, and Biocompatibility Potential of Aqueous Zingiber officinale Rhizome (AZOME) Extract

Mohd Hasan Mujahid ¹, Tarun Kumar Upadhyay ^1*, Vijay Jagdish Upadhye ^2*, Prasanna Sriram Mathad ³

This study determined the therapeutic potential of Aqueous Zingiber officinale rhizome extract (AZOME) in combating oxidative stress, microbial infections, diabetes, and glycation-related complications.

Abstract

Background: The rhizome of Zingiber officinale or ginger shows its potential against oxidative stress, microbial infections, and managing diabetes mellitus. Method: Chemical reagents and plant materials of analytical grade were procured. Aqueous extract of Zingiber officinale rhizome was prepared through maceration. FT-IR, heavy metal detection, UV-VIS spectroscopy, and carbohydrate estimation were performed. Antimicrobial activity against bacterial and fungal strains was evaluated using agar diffusion method. Total polyphenol, flavonoid, and flavonol contents were quantified. Antioxidant activity was assessed using DPPH, reducing power, and FRAP assays. Antidiabetic activity was determined by α-amylase and α-glucosidase inhibition. Cytotoxicity, glucose uptake, and antiglycation assays were conducted on L6 cells. Blood compatibility was tested on human RBCs. Results: The analysis showed total phenolic content (TPC) at 27.9±0.27 mg/g GAE, total flavonoid content (TFC) at 18.4 mg/g QE, and total flavonol content (TFolC) at 41.1±4 mg/g QE. The extract demonstrated potent antioxidant activity with IC50 values of 353 µg/mL (DPPH), 600 µg/mL (H2O2 scavenging), and displayed antidiabetic effects inhibiting α-amylase (IC50=1564.43 µg/mL) and α-glucosidase (IC50=581.4 µg/mL). Cytotoxicity assays yielded an IC50 of 533.3 µg/mL (NRU assay) and highest glucose uptake at 254.74±62.79 µg/mL (L6 cells). The extract showed minimal hemolytic activity (-0.305±0.031%) and high cell viability (99%). In silico docking revealed strong interactions with targeted proteins. Conclusion: In conclusion, the aqueous extract of Zingiber officinale rhizome exhibits diverse pharmacological activities including antioxidant, antimicrobial, antidiabetic, and antiglycation properties, along with notable bio-safety and cytotoxicity profiles.

Keywords: Zingiber officinale, Ginger, Antioxidant, Antimicrobial, Antidiabetic, Antiglycation, Biocompatibility, In silico

References

Abdullah, Hussain T, Faisal S, Rizwan M, Almostafa MM, Younis NS, Yahya G (2023) Zingiber officinale rhizome extracts mediated ni nanoparticles and its promising biomedical and environmental applications. BMC Complementary Medicine and Therapies 23 (1):349. https://doi.org/10.1186/s12906-023-04182-7

Adeyeoluwa TE, Balogun FO, Ashafa AOT (2020) In vitro comparative assessment of the inhibitory effects of single and combined spices against glucose-synthesizing enzymes. Tropical Journal of Pharmaceutical Research 19 (6):1209-1214. https://doi.org/10.4314/tjpr.v19i6.14

Akbari S, Abdurahman NH, Yunus RM, Alara OR, Abayomi OO (2019) Extraction, characterization and antioxidant activity of fenugreek (Trigonella-Foenum Graecum) seed oil. Materials Science for Energy Technologies 2 (2):349-355. https://doi.org/10.1016/j.mset.2018.12.001

Al-Mustafa A, Al-Tawarah M, Al-Sheraideh MS, Al-Zahrany FA (2021) Phytochemical analysis, antioxidant and in vitro β-galactosidase inhibition activities of Juniperus phoenicea and Calicotome villosa methanolic extracts. BMC chemistry 15:1-13. https://doi.org/10.1186/s13065-021-00781-y

Ali A, Gilani AH (2007) Medicinal value of ginger with focus on its use in nausea and vomiting of pregnancy. International Journal of Food Properties 10 (2):269-278. https://doi.org/10.1080/10942910601045297

Ali BH, Blunden G, Tanira MO, Nemmar A (2008) Some phytochemical, pharmacological and toxicological properties of ginger (Zingiber officinale Roscoe): a review of recent research. Food and chemical Toxicology 46 (2):409-420.https://doi.org/10.1016/j.fct.2007.09.085

Alsahli MA, Almatroodi SA, Almatroudi A, Khan AA, Anwar S, Almutary AG, Alrumaihi F, Rahmani AH (2021) 6-Gingerol, a major ingredient of ginger attenuates diethylnitrosamine-induced liver injury in rats through the modulation of oxidative stress and anti-inflammatory activity. Mediators of inflammation 2021:1-17. https://doi.org/10.1155/2021/6661937 33

Alvi SS, Nabi R, Khan M, Akhter F, Ahmad S, Khan MS (2021) Glycyrrhizic acid scavenges reactive carbonyl species and attenuates glycation-induced multiple protein modification: an in vitro and in silico study. Oxidative Medicine and Cellular Longevity 2021. https://doi.org/10.1155/2021/7086951

Ashraf JM, Rabbani G, Ahmad S, Hasan Q, Khan RH, Alam K, Choi I (2015) Glycation of H1 histone by 3-deoxyglucosone: effects on protein structure and generation of different advanced glycation end products. PLoS One 10 (6):e0130630. https://doi.org/10.1371/journal.pone.0130630

Bahuguna A, Khan I, Bajpai VK, Kang SC (2017) MTT assay to evaluate the cytotoxic potential of a drug. ||| Bangladesh Journal of Pharmacology 12 (2):115-118. https://doi.org/10.3329/bjp.v12i2.30892

Banday MZ, Sameer AS, Nissar S (2020) Pathophysiology of diabetes: An overview. Avicenna journal of medicine 10 (04):174-188. https://doi.org/10.4103/ajm.ajm_53_20

Beristain-Bauza SDC, Hernández-Carranza P, Cid-Pérez TS, Ávila-Sosa R, Ruiz-López II, Ochoa-Velasco CE (2019) Antimicrobial activity of ginger (Zingiber officinale) and its application in food products. Food Reviews International 35 (5):407-426. https://doi.org/10.1080/87559129.2019.1573829

Eid BG, Mosli H, Hasan A, El-Bassossy HM (2017) Ginger ingredients alleviate diabetic prostatic complications: Effect on oxidative stress and fibrosis. Evidence-based complementary and alternative medicine 2017. https://doi.org/10.1155/2017/6090269

Fernandes RdPP, Trindade MA, Tonin FG, Lima CGd, Pugine SMP, Munekata PES, Lorenzo J, De Melo M (2016) Evaluation of antioxidant capacity of 13 plant extracts by three different methods: cluster analyses applied for selection of the natural extracts with higher antioxidant capacity to replace synthetic antioxidant in lamb burgers. Journal of food science and technology 53:451-460. https://doi.org/10.1007/s13197-015-1994-x

Gohel A, Upadhye V, K Upadhyay T, Rami E, Panchal R, Jadhav S, Dhakane R, Kele V (2021) Study on phytochemical screening and antimicrobial activity of Adhatoda vasica. Canadian Journal of Medicine 3 (3):105-113. https://doi.org/10.33844/cjm.2021.60509

Hafeez J, Naeem M, Ali T, Sultan B, Hussain F, Ur Rashid H, Nadeem M, Shirzad I (2023) Comparative Study of Antioxidant, Antidiabetic, Cytotoxic Potentials, and Phytochemicals of Fenugreek (Trigonella foenum-graecum) and Ginger (Zingiber officinale). Journal of Chemistry 2023. https://doi.org/10.1155/2023/3469727

Hasan Mujahid M, Upadhyay TK, Upadhye V, Sharangi AB, Saeed M (2023) Phytocompound identification of aqueous Zingiber officinale rhizome (ZOME) extract reveals antiproliferative and reactive oxygen species mediated apoptotic induction within cervical cancer cells: an in vitro and in silico approach. Journal of Biomolecular Structure and Dynamics:1-28. https://doi.org/10.1080/07391102.2023.2247089

Hosseini A, Alavi MS, Toos MGN, Jamialahmadi T, Sahebkar A (2024) 6-Gingerol, an ingredient of Zingiber officinale, abrogates lipopolysaccharide-induced cardiomyocyte injury by reducing oxidative stress and inflammation. Journal of Agriculture and Food Research 15:101034. https://doi.org/10.1016/j.jafr.2024.101034

Husen A (2023) Antidiabetic medicinal plants and herbal treatments. CRC Press. https://doi.org/10.1201/b23347

Huyut Z, Beydemir S, Gülçin I (2017) Antioxidant and antiradical properties of selected flavonoids and phenolic compounds. Biochemistry research international 2017. https://doi.org/10.1155/2017/7616791

Jiang C, Wang L, Shao J, Jing H, Ye X, Jiang C, Wang H, Ma C (2020) Screening and identifying of α-amylase inhibitors from medicine food homology plants: Insights from computational analysis and experimental studies. Journal of Food Biochemistry 44 (12):e13536. https://doi.org/10.1111/jfbc.13536

Jung H, Kim YS, Jung D-M, Lee K-S, Lee J-M, Kim KK (2022) Melittin-derived peptides exhibit variations in cytotoxicity and antioxidant, anti-inflammatory and allergenic activities. Animal Cells and Systems 26 (4):158-165. https://doi.org/10.1080/19768354.2022.2099971

Khan F, Pandey P, Singh A, Upadhyay TK, AboElnaga SMH, Al-Najjar MA, Saeed M, Kahrizi D (2022) Unveiling Antioxidant and Antiproliferative Effects of Prosopis juliflora Leaves against Human Prostate Cancer LNCaP Cells. Cellular and Molecular Biology 68 (11):20-27. https://doi.org/10.14715/cmb/2022.68.11.4

Kumar A, Gangwar R, Ahmad Zargar A, Kumar R, Sharma A (2024) Prevalence of diabetes in India: A review of IDF diabetes atlas 10th edition. Current diabetes reviews 20 (1):105-114. https://doi.org/10.2174/1573399819666230413094200

Kurhekar JV (2021) Ancient and modern practices in phytomedicine. In:  Preparation of Phytopharmaceuticals for the Management of Disorders. Elsevier, pp 55-75. https://doi.org/10.1016/b978-0-12-820284-5.00019-8

Kusse Gudishe Goroya KGG, Zewde Mitiku ZM, Yoseph Alresawum Asresahegn YAA (2019) Determination of concentration of heavy metals in ginger using flame atomic absorption spectroscopy.https://doi.org/10.5897/ajps2019.1787

Liu Y, Yang X, Gan J, Chen S, Xiao Z-X, Cao Y (2022) CB-Dock2: Improved protein–ligand blind docking by integrating cavity detection, docking and homologous template fitting. Nucleic acids research 50 (W1):W159-W164. https://doi.org/10.1093/nar/gkac394

Ma R-H, Ni Z-J, Zhu Y-Y, Thakur K, Zhang F, Zhang Y-Y, Hu F, Zhang J-G, Wei Z-J (2021) A recent update on the multifaceted health benefits associated with ginger and its bioactive components. Food & Function 12 (2):519-542. https://doi.org/10.1039/d0fo02834g

Mandal SP, Garg A, Prabitha P, Wadhwani AD, Adhikary L, Kumar BP (2018) Novel glitazones as PPARγ agonists: molecular design, synthesis, glucose uptake activity and 3D QSAR studies. Chemistry Central Journal 12:1-21. https://doi.org/10.1186/s13065-018-0508-0

Mohammadinejad R, Shavandi A, Raie DS, Sangeetha J, Soleimani M, Hajibehzad SS, Thangadurai D, Hospet R, Popoola JO, Arzani A (2019) Plant molecular farming: production of metallic nanoparticles and therapeutic proteins using green factories. Green chemistry 21 (8):1845-1865. https://doi.org/10.1039/c9gc00335e

Mohd Dom NS, Yahaya N, Adam Z, Hamid M (2020) Antiglycation and antioxidant properties of Ficus deltoidea varieties. Evidence-Based Complementary and Alternative Medicine 2020. https://doi.org/10.1155/2020/6374632

Nabi R, Alvi SS, Khan RH, Ahmad S, Ahmad S, Khan MS (2018) Antiglycation study of HMG-R inhibitors and tocotrienol against glycated BSA and LDL: A comparative study. International journal of biological macromolecules 116:983-992. https://doi.org/10.1016/j.ijbiomac.2018.05.115

Nabi R, Alvi SS, Saeed M, Ahmad S, Khan MS (2019) Glycation and HMG-CoA reductase inhibitors: implication in diabetes and associated complications. Current diabetes reviews 15 (3):213-223. https://doi.org/10.2174/1573399814666180924113442

Nabi R, Alvi SS, Shah MS, Ahmad S, Faisal M, Alatar AA, Khan MS (2020) A biochemical & biophysical study on in-vitro anti-glycating potential of iridin against D-Ribose modified BSA. Archives of biochemistry and biophysics 686:108373. https://doi.org/10.1016/j.abb.2020.108373

Nariya PB, Bhalodia NR, Shukla V, Acharya R (2011) Antimicrobial and antifungal activities of Cordia dichotoma (Forster F.) bark extracts. AYU (An international quarterly journal of research in Ayurveda) 32 (4):585-589. https://doi.org/10.4103/0974-8520.96138

Nguyen ST, Vo PH, Nguyen TD, Do NM, Le BH, Dinh DT, Truong KD, Van Pham P (2019) Ethanol extract of Ginger Zingiber officinale Roscoe by Soxhlet method induces apoptosis in human hepatocellular carcinoma cell line. Biomedical Research and Therapy 6 (11):3433-3442. https://doi.org/10.15419/bmrat.v6i11.572

Noipha K, Ninla-Aesong P (2018) Antidiabetic activity of Zingiber officinale Roscoe rhizome extract: an in vitro study. HAYATI Journal of Biosciences 25 (4):160-160. https://doi.org/10.4308/hjb.25.4.160

Offei-Oknye R, Patterson J, Walker L, Verghese M (2015) Processing effects on phytochemical content and antioxidative potential of ginger Zingiber officale. Food and Nutrition Sciences 6 (5):445-451. https://doi.org/10.4236/fns.2015.65046

Osabor V, Bassey F, Umoh U (2015) Phytochemical screening and quantitative evaluation of nutritional values of Zingiber officinale (Ginger). American Chemical Science Journal 8 (4):1-6. https://doi.org/10.9734/acsj/2015/16915

Oyedemi S, Bradley G, Afolayan A (2012) In vitro and in vivo antioxidant properties of aqueous extract from Strychnos henningsii Gilg stem bark. Free Radical Biology and Medicine (53):S113. https://doi.org/10.1016/j.freeradbiomed.2012.08.237

Oyedemi SO, Oyedemi BO, Ijeh II, Ohanyerem PE, Coopoosamy RM, Aiyegoro OA (2017) Alpha-amylase inhibition and antioxidative capacity of some antidiabetic plants used by the traditional healers in Southeastern Nigeria. The Scientific World Journal 2017. https://doi.org/10.1155/2017/3592491

Papoutsis K, Zhang J, Bowyer MC, Brunton N, Gibney ER, Lyng J (2021) Fruit, vegetables, and mushrooms for the preparation of extracts with α-amylase and α-glucosidase inhibition properties: A review. Food Chemistry 338:128119. https://doi.org/10.1016/j.foodchem.2020.128119

Rhee SY, Kim YS (2018) The role of advanced glycation end products in diabetic vascular complications. Diabetes & metabolism journal 42 (3):188. https://doi.org/10.4093/dmj.2017.0105

Richards C, O'Connor N, Jose D, Barrett A, Regan F (2020) Selection and optimization of protein and carbohydrate assays for the characterization of marine biofouling. Analytical methods 12 (17):2228-2236. https://doi.org/10.1039/d0ay00272k

Sahoo MR, Umashankara MS (2023) FTIR Based metabolomics profiling and fingerprinting of some medicinal plants: an attempt to develop an approach for quality control and standardization of herbal materials. Pharmacognosy Research 15 (1). https://doi.org/10.5530/097484900288

Salehi B, Martorell M, Arbiser JL, Sureda A, Martins N, Maurya PK, Sharifi-Rad M, Kumar P, Sharifi-Rad J (2018) Antioxidants: positive or negative actors? Biomolecules 8 (4):124. https://doi.org/10.3390/biom8040124

Saraf S (2012) Development of Fingerprinting Methods of Balacaturbhadrika Churna: An Ayurvedic Formulation. Pharmacognosy Journal 4 (27):20-24. https://doi.org/10.5530/pj.2012.27.3

Shah A, Niaz A, Ullah N, Rehman A, Akhlaq M, Zakir M, Suleman Khan M (2013) Comparative study of heavy metals in soil and selected medicinal plants. Journal of Chemistry 2013. https://doi.org/10.1155/2013/621265

Sivasothy Y, Chong WK, Hamid A, Eldeen IM, Sulaiman SF, Awang K (2011) Essential oils of Zingiber officinale var. rubrum Theilade and their antibacterial activities. Food chemistry 124 (2):514-517. https://doi.org/10.1016/j.foodchem.2010.06.062

Slowing II, Wu CW, Vivero-Escoto JL, Lin VSY (2009) Mesoporous silica nanoparticles for reducing hemolytic activity towards mammalian red blood cells. Small 5 (1):57-62. https://doi.org/10.1002/smll.200800926

Srinivasan K (2017) Ginger rhizomes (Zingiber officinale): A spice with multiple health beneficial potentials. PharmaNutrition 5 (1):18-28. https://doi.org/10.1016/j.phanu.2017.01.001

Tareen AK, Panezai MA, Sajjad A, Achakzai JK, Kakar AM, Khan NY (2021) Comparative analysis of antioxidant activity, toxicity, and mineral composition of kernel and pomace of apricot (Prunus armeniaca L.) grown in Balochistan, Pakistan. Saudi Journal of Biological Sciences 28 (5):2830-2839. https://doi.org/10.1016/j.sjbs.2021.02.015

Telagari M, Hullatti K (2015) In-vitro α-amylase and α-glucosidase inhibitory activity of Adiantum caudatum Linn. and Celosia argentea Linn. extracts and fractions. Indian journal of pharmacology 47 (4):425-429. https://doi.org/10.4103/0253-7613.161270

Trott O, Olson AJ (2010) AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. Journal of computational chemistry 31 (2):455-461. https://doi.org/10.1002/jcc.21334

Upadhyay TK, Das S, Mathur M, Alam M, Bhardwaj R, Joshi N, Sharangi AB (2024) Medicinal plants and their bioactive components with antidiabetic potentials. Antidiabetic Medicinal Plants:327-364. https://doi.org/10.1016/b978-0-323-95719-9.00017-3

Vajrabhaya L-O, Korsuwannawong S (2016) Cytotoxicity evaluation of Clinacanthus nutans through dimethylthiazol diphenyltetrazolium bromide and neutral red uptake assays. European journal of dentistry 10 (01):134-138. https://doi.org/10.4103/1305-7456.175701

Valdés-Tresanco MS, Valdés-Tresanco ME, Valiente PA, Moreno E (2020) AMDock: a versatile graphical tool for assisting molecular docking with Autodock Vina and Autodock4. Biology direct 15:1-12. https://doi.org/10.1186/s13062-020-00267-2

Vishwanath D, Srinivasan H, Patil MS, Seetarama S, Agrawal SK, Dixit M, Dhar K (2013) Novel method to differentiate 3T3 L1 cells in vitro to produce highly sensitive adipocytes for a GLUT4 mediated glucose uptake using fluorescent glucose analog. Journal of cell communication and signaling 7 (2):129-140. https://doi.org/10.1007/s12079-012-0188-9

Visvanathan R, Jayathilake C, Liyanage R (2016) A simple microplate-based method for the determination of α-amylase activity using the glucose assay kit (GOD method). Food chemistry 211:853-859. https://doi.org/10.1016/j.foodchem.2016.05.090

Warren FJ, Zhang B, Waltzer G, Gidley MJ, Dhital S (2015) The interplay of α-amylase and amyloglucosidase activities on the digestion of starch in in vitro enzymic systems. Carbohydrate polymers 117:192-200. https://doi.org/10.1016/j.carbpol.2014.09.043

Yousfi F, Abrigach F, Petrovic J, Sokovic M, Ramdani M (2021) Phytochemical screening and evaluation of the antioxidant and antibacterial potential of Zingiber officinale extracts. South African Journal of Botany 142:433-440. https://doi.org/10.1016/j.sajb.2021.07.010

Zheng Q, Cheng Z, Duan Y, Hu K, Cai M, Zhang H (2024) Effect of subcritical water temperature on the chain conformation and immune activity of ginger polysaccharides. International Journal of Biological Macromolecules 261:129833. https://doi.org/10.1016/j.ijbiomac.2024.129833