Flash chromatographic separation and diabetic wound healing study of Hemigraphis alternata leaves extract

Year 2025, Volume: 29 Issue: 2, 605 - 615

Asha Thomas , Namita Surve Pranali Jadhav , Vrushali Balchim Lata Kothapalli , Sohan Chitlange

https://doi.org/10.12991/jrespharm.1662192

Abstract

Diabetes, a multifaceted disorder associated with delayed wound healing due to persistent hyperglycaemia and related comorbidities, which increases morbidity, mortality and worsens quality of life. Hemigraphis alternata possess wound healing, antioxidant, anti-inflammatory and anti-diabetic properties. This study was conducted to assess the efficacy of H. alternata leaf extract in the treatment of diabetic wounds. Hydroalcoholic extract from H. alternata leaves was cold-extracted and fractionated via flash chromatography. Collected fractions underwent phytochemical screening for active constituents. An emulgel with the extract was developed and tested for wound healing in STZ + Nicotinamide induced diabetic rats.Flash chromatographic separation of extract yielded 12 fractions and its phytochemical screening showed presence of various phytoconstituents such as alkaloids, phenols, flavonoids, terpenoids, etc which might responsible for wound healing. Wound closure rate in rats treated with formulated emulgel was significantly faster than the untreated diabetic rats. The emulgel group showed low level pro-inflammatory cytokines (IL-6 and TNF-α) and high concentration of VEGF. This study has shown that H. alternata emulgel has a significant potential in diabetic wound management by enhancing the release of VEGF and reducing the production of pro-inflammatory cytokines which induces cell proliferation, migration and wound contraction.

Keywords

Diabetic wound healing, Flash chromatography, Emulgel, Hemigraphis alternata, Pro-inflammatory cytokines, Diabetes

References

• [1] Cheng KY, Lin ZH, Cheng YP, Chiu HY, Yeh NL, Wu TK, Wu JS. Wound healing in streptozotocin-ınduced diabetic rats using atmospheric-pressure argon plasma jet. Sci Rep. 2018; 8(1): 12214. https://doi.org/10.1038/s41598-018-30597-1
• [2] Jeffcoate WJ, Price P, Harding KG. Wound healing and treatments for people with diabetic foot ulcers. Diabetes Metab Res Rev. 2004; 20(l1): S78-89. https://doi.org/10.1002/dmrr.476
• [3] Gao D, Zhang Y, Bowers DT, Liu W, Ma M. Functional hydrogels for diabetic wound management. APL Bioeng. 2021; 5(3): 031503. https://doi.org/10.1063/5.0046682
• [4] Patel S, Srivastava S, Singh MR, Singh D. Mechanistic insight into diabetic wounds: Pathogenesis, molecular targets and treatment strategies to pace wound healing. Biomed Pharmacother. 2019; 112: 108615. https://doi.org/10.1016/j.biopha.2019.108615
• [5] Rekha PD, Rao SS, Sahana TG, Prabhu A. Diabetic wound management. Br J Community Nurs. 2018; 23(9): S16-S22. https://doi.org/10.12968/bjcn.2018.23.sup9.s16
• [6] Xu F, Zhang C, Graves DT. Abnormal cell responses and role of TNF-α in impaired diabetic wound healing. Biomed Res Int. 2013; 2013: 754802. https://doi.org/10.1155/2013/754802
• [7] Dixon D, Edmonds M. Managing diabetic foot ulcers: Pharmacotherapy for wound healing. Drugs. 2021; 81(1): 29-56. https://doi.org/10.1007/s40265-020-01415-8
• [8] Shah SA, Sohail M, Khan S, Minhas MU, de Matas M, Sikstone V, Hussain Z, Abbasi M, Kousar M. Biopolymer-based biomaterials for accelerated diabetic wound healing: A critical review. Int J BiolMacromol. 2019; 139: 975-993. https://doi.org/10.1016/j.ijbiomac.2019.08.007
• [9] Kashyap AK, Reddy NP, Chaitanya RK, Karnati R. Ethyl acetate extract of Hemigraphis colorata leaves shows anti-inflammatory and wound healing properties and inhibits 5-lipoxygenase and cyclooxygenase-1 and 2 enzymes. J Med Plant Res. 2013; 7(37): 2783-2791. https://doi.org/10.5897/JMPR2013.5184
• [10] Priya MD. Review on pharmacological activity of Hemigraphis colorata (Blume). Int J Herb Med. 2013; 1(3): 120-121.
• [11] Edwin BT, Nair PD. In vitro evaluation of wound healing property of Hemigraphis alternata (Burm. F) T. Anders using fibroblast and endothelial cells. Biosci Biotechnol Res Asis. 2011; 8(1): 185-193. https://www.biotech-asia.org/?p=9316
• [12] Akhil TT, Rangasamy KS, Nadu T. Evaluation of anti-oxidant, anti-inflammatory and cytotoxicity potential of Hemigraphs colorata. Int J Pharm Sci. 2013; 4(9): 3477-3483. http://dx.doi.org/10.13040/IJPSR.0975-8232.4(9).3477-83
• [13] Roy S, Zomuanpuia JH, Thangliani L, Malsawmdawngkimi RL, Hulianthanga, Lalhriatzuali J, Hauhnar L, Lalchhandama K, Lalhminghlui K, Lalthanpuii PB. Screening of Callicarpa arborea and Hemigraphis alternata for antibacterial activity. Sci Vis. 2020; 20(2): 72-77. http://dx.doi.org/10.33493/scivis.20.02.02
• [14] Comino-Sanz IM, López-Franco MD, Castro B, Pancorbo-Hidalgo PL. The role of antioxidants on wound healing: A review of the current evidence. J Clin Med. 2021; 10(16): 3558. https://doi.org/10.3390/jcm10163558
• [15] Baba SA, Malik SA. Determination of total phenolic and flavonoid content, antimicrobial and antioxidant activity of a root extract of Arisaema jacquemontii Blume. Integr Med Res. 2015; 9(4): 449-454. http://dx.doi.org/10.1016/j.jtusci.2014.11.001
• [16] Bhaigyaba T, Bag GC, Grihanjali DP. Assessment of total flavonoid content and antioxidant activity of methanolic rhizome extract of three Hedychium species of Manipur valley. Int J Pharm Sci Rev Res. 2015; 30(1): 154-159.
• [17] Rahman MM, Islam MB, Biswas M, Khurshid Alam AH. In vitro antioxidant and free radical scavenging activity of different parts of Tabebuia pallida growing in Bangladesh. BMC Res Notes. 2015; 8: 621. https://doi.org/10.1186/s13104-015-1618-6
• [18] Chromatography HT. Procedure for Identification of articles of botanical origin preparation of test solution. 2015: 1-2.
• [19] Khullar R, Kumar D, Seth N, Saini S. Formulation and evaluation of mefenamic acid emulgel for topical delivery. Saudi Pharm J. 2012; 20(1): 63-67. https://doi.org/10.1016/j.jsps.2011.08.001
• [20] Arshad W, MuhammadH, Khan S, Akhtar N. Polymeric emulgel carrying Cinnamomum tamala extract: Promising delivery system for potential topical applications. Braz J Pharm Sci. 2011: 1-11. https://doi.org/10.1590/s2175-97902019000418318
• [21] Kumar K, Kumar N, Singh A, Gupta A. Development and evaluation of curcumin emulgel with cumin oil for topical application. Clin Pharmacol Biopharm. 2021; 10: 233. https://doi.org/10.4172/2167-065X.1000233
• [22] Burki IK, Khan MK, Khan BA, Uzair B, Braga VA, Jamil QA. Formulation development, characterization, and evaluation of a novel dexibuprofen-capsaicin skin emulgel with improved in vivo anti-inflammatory and analgesic effects. AAPS Pharm Sci Tech. 2020; 21(6): 211. https://doi.org/10.1208/s12249-020-01760-7
• [23] OECD 404. Guidelines for the testing of chemicals. Acute Dermal Irritation/Corrosion. OECD Guidel Test Chem Sect 4 Heal Eff. 2015: 1-8. https://www.oecd.org/env/test-no-404-acute-dermal-irritation-corrosion-9789264242678-en.htm (Accessed on 30 March, 2024)
• [24] Nahid MHE, ShadiaAO, Hala AMI. Histopathological alteration in stz-nicotinamide diabetic rats, a complication of diabetes or a toxicity of ATZ? Int J Diabetes Clin Res. 2018: 5(3). https://doi.org/10.23937/2377-3634/1410091
• [25] Hashemnia M, Nikousefat Z, Mohammadalipour A, Zangeneh MM, Zangeneh A. Wound healing activity of Pimpinella anisum methanolic extract in streptozotocin-induced diabetic rats. J Wound Care. 2019; 28(10): S26-S36. https://doi.org/10.12968/jowc.2019.28.sup10.s26
• [26] Grada A, Mervis J, Falanga V. Research techniques made simple: Animal models of wound healing. J Invest Dermatol. 2018; 138(10): 2095-2105. https://doi.org/10.1016/j.jid.2018.08.005
• [27] Alizargar J, Kuchaki E, Shaabani A, Namazi M. Properties of wound healing activities of rosemary extract. J Biol Act Prod Nat. 2012; 2(4): 218-224. http://dx.doi.org/10.1080/22311866.2012.10719128
• [28] Sanapalli BKR, Yele V, Singh MK, Thaggikuppe Krishnamurthy P, Karri VVSR. Preclinical models of diabetic wound healing: A critical review. Biomed Pharmacother. 2021; 142: 111946. https://doi.org/10.1016/j.biopha.2021.111946
• [29] Prasad SK, Kumar R, Patel DK, Hemalatha S. Wound healing activity of Withania coagulans in streptozotocin-induced diabetic rats. Pharm Biol. 2010; 48(12): 1397-1404. https://doi.org/10.3109/13880209.2010.486837