Adjunctive Nano-Formulated Kratom (Mytragina Speciosa) with Paracetamol for Neuropathic Pain in Rats: Mechanical Allodynia and Serum Interleukin-6 Outcomes
-
Rahmat Kukuh Noviandomo
,
Anna Surgean Veterini
,
Kohar Hari Santosa
,
Prananda Surya Airlangga
,
Christrijogo Sumartono Waloejo
Abstract
Background: Neuropathic pain is a refractory chronic pain condition driven in part by neuroinflammation. Interleukin-6 (IL-6) is a key pro-inflammatory mediator, and paracetamol monotherapy is often insufficient. Alkaloids of Mitragyna speciosa (kratom) have μ-opioid receptor agonist and anti-inflammatory activity, while nanoformulation may improve oral bioavailability. The objective is to evaluate the analgesic and anti-inflammatory effects of oral nano-kratom as an adjuvant to paracetamol on mechanical withdrawal threshold and serum IL-6 in a rat chronic constriction injury (CCI) model.
Methods: A post-test-only randomized controlled animal study was conducted in 21 male Wistar rats assigned to vehicle control (K−), paracetamol 10 mg/kg (K1), or nano-kratom 30 mg/kg plus paracetamol 5 mg/kg (K2) (n = 7/group). After acclimatization, unilateral CCI of the left sciatic nerve was induced. Treatments were given orally once daily from day 7 to day 13, and endpoints were assessed on day 14. Mechanical withdrawal threshold was measured using von Frey filaments, and serum IL-6 was quantified by ELISA. One K− rat died before endpoint assessment; the final analyzed n = 6, 7, and 7.
Results: Baseline von Frey thresholds were comparable across groups. CCI significantly reduced withdrawal thresholds in all groups. On day 14, thresholds differed significantly, with K2 showing the highest values, followed by K1 and K− (all pairwise p < 0.001). Serum IL-6 was lowest in K2; the difference was significant versus K− (p = 0.037) and approached significance versus K1 (p = 0.053).
Conclusion: Nano-kratom combined with half-dose paracetamol produced greater antinociception than paracetamol alone and was associated with the lowest serum IL-6 levels. These findings support further evaluation of nano-formulated kratom as a multimodal adjuvant for neuropathic pain.
[1] Clark AK, Old EA, Malcangio M. Neuropathic pain and cytokines: current perspectives. J Pain Res. 2013; 6:803-14.
[2] Colloca L, Ludman T, Bouhassira D, Baron R, Dickenson AH, Yarnitsky D, et al. Neuropathic pain. Nat Rev Dis Primers. 2017; 3:17002.
[3] Zhou YQ, Liu Z, Liu ZH, Chen SP, Li M, Shahveranov A, et al. Interleukin-6: an emerging regulator of pathological pain. J Neuroinflammation. 2016; 13(1):141.
[4] Lee KM, Jeon SM, Cho HJ. Interleukin-6 induces microglial CX3CR1 expression in the spinal cord after peripheral nerve injury through the activation of p38 MAPK. Eur J Pain. 2010; 14(7):682.e1-12.
[5] Scheller J, Garbers C, Rose-John S. Interleukin-6: from basic biology to selective blockade of pro-inflammatory activities. Semin Immunol. 2014; 26(1):2-12.
[6] Karunakaran T, Ngew KZ, Zailan AAD, Mian Jong VY, Abu Bakar MH. The chemical and pharmacological properties of mitragynine and its diastereomers: an insight review. Front Pharmacol. 2022; 13:805986.
[7] Kruegel AC, Gassaway MM, Kapoor A, Váradi A, Majumdar S, Filizola M, et al. Synthetic and receptor signaling explorations of the Mitragyna alkaloids: mitragynine as an atypical molecular framework for opioid receptor modulators. J Am Chem Soc. 2016; 138(21):6754-64.
[8] Prozialeck WC, Jivan JK, Andurkar SV. Pharmacology of kratom: an emerging botanical agent with stimulant, analgesic and opioid-like effects. J Am Osteopath Assoc. 2012; 112(12):792-9.
[9] Azhar S, Ashari S. Strategies for the development of Mitragyna speciosa (kratom) leaves extract loaded with solid lipid nanoparticles. JMEST. 2019; 6(12):100-4.
[10] Azhar SN, Ashari SE, Tan JK, Kassim NK, Hassan M, Zainuddin N, et al. Screening and selection of formulation components of nanostructured lipid carriers system for Mitragyna speciosa (Korth). Havil drug delivery. Ind Crops Prod. 2023; 198:116668.
[11] Peng Y, Chen L, Ye S, Kang Y, Liu J, Zeng S, et al. Research and development of drug delivery systems based on drug transporter and nano-formulation. Asian J Pharm Sci. 2020; 15(2):220-36.
[12] Dubin AE, Patapoutian A. Nociceptors: the sensors of the pain pathway. J Clin Invest. 2010; 120(11):3760-72.
[13] Cavalli E, Mammana S, Nicoletti F, Bramanti P, Mazzon E. The neuropathic pain: an overview of the current treatment and future therapeutic approaches. Int J Immunopathol Pharmacol. 2019; 33:2058738419838383.
[14] Bouhassira D, Lantéri-Minet M, Attal N, Laurent B, Touboul C. Prevalence of chronic pain with neuropathic characteristics in the general population. Pain. 2008; 136(3):380-7.
[15] van Hecke O, Austin SK, Khan RA, Smith BH, Torrance N. Neuropathic pain in the general population: a systematic review of epidemiological studies. Pain. 2014; 155(9):1907.
[16] Torrance N, Smith BH, Bennett MI, Lee AJ. The epidemiology of chronic pain of predominantly neuropathic origin. Results from a general population survey. J Pain. 2006; 7(4):281-9.
[17] Zimmer Z, Fraser K, Grol-Prokopczyk H, Zajacova A. A global study of pain prevalence across 52 countries: examining the role of country-level contextual factors. Pain. 2022; 163(9):1740-50.
[18] Purwata TE, Sadeli HA, Amir D, Asnawi C, Partoatmodjo L, Aulina S, et al. Characteristics of neuropathic pain in Indonesia: a hospital based national clinical survey. Neurol Asia. 2015; 20(4).
[19] Dureja GP, Iyer RN, Das G, Ahdal J, Narang P. Evidence and consensus recommendations for the pharmacological management of pain in India. J Pain Res. 2017; 10:709-36.
[20] Gilron I, Baron R, Jensen T. Neuropathic pain: principles of diagnosis and treatment. Mayo Clin Proc. 2015; 90(4):532-45.
[21] Wright ME, Rizzolo D. An update on the pharmacologic management and treatment of neuropathic pain. JAAPA. 2017; 30(3):13-7.
[22] Vargas-Schaffer G. Is the WHO analgesic ladder still valid? Twenty-four years of experience. Can Fam Physician. 2010; 56(6):514-7.
[23] Dani M, Guindon J, Lambert C, Beaulieu P. The local antinociceptive effects of paracetamol in neuropathic pain are mediated by cannabinoid receptors. Eur J Pharmacol. 2007; 573(1-3):214-5.
[24] Gulcu N, Karaaslan K, Ozturan KE, Kocoglu H. Intravenous paracetamol for pain relief in a patient with neuropathic pain. Acta Anaesthesiol Scand. 2007; 51(7):956-7.
[25] Mazaleuskaya LL, Sangkuhl K, Thorn CF, FitzGerald GA, Altman RB, Klein TE. PharmGKB summary: pathways of acetaminophen metabolism at the therapeutic versus toxic doses. Pharmacogenet Genomics. 2015; 25(8):416-26.
[26] Ohashi N, Kohno T. Analgesic effect of acetaminophen: a review of known and novel mechanisms of action. Front Pharmacol. 2020; 11:580289.
[27] Im KS, Jung HJ, Kim JB, Lee JM, Park HJ, Joo CH, et al. The antinociceptive effect of acetaminophen in a rat model of neuropathic pain. Kaohsiung J Med Sci. 2012; 28(5):251-8.
[28] Beiranvand S, Sorori MM. Pain management using nanotechnology approaches. Artif Cells Nanomed Biotechnol. 2019; 47(1):462-8.
[29] Gunn J, Hill MM, Cotten BM, Deer TR. An analysis of biomarkers in patients with chronic pain. Pain Physician. 2020; 23(2):235.
[30] Persson AK, Gebauer M, Jordan S, Metz-Weidmann C, Schulte AM, Schneider HC, et al. Correlational analysis for identifying genes whose regulation contributes to chronic neuropathic pain. Mol Pain. 2009; 5:7.
[31] Assanangkornchai S, Muekthong A, Sam-Angsri N, Pattanasattayawong U. The use of Mitragyna speciosa ("krathom"), an addictive plant, in Thailand. Subst Use Misuse. 2007; 42(14):2145-57.
[32] Wahyono S, Widowati L, Handayani L, Sampurno OD, Haryanti S, Fauzi F, et al. Kratom, prospek kesehatan dan sosial ekonomi. Jakarta: Lembaga Penerbit Badan Penelitian dan Pengembangan Kesehatan; 2019.
[33] Stolt AC, Schröder H, Neurath H, Grecksch G, Höllt V, Meyer MR, et al. Behavioral and neurochemical characterization of kratom (Mitragyna speciosa) extract. Psychopharmacology (Berl). 2014; 231(1):13-25.
[34] Anantaworasakul P, Preedalikit W, Anantaworasakul P, Singh S, Intharuksa A, Arunotayanun W, et al. Phytochemical characterization, bioactivities, and nanoparticle-based topical gel formulation development from four Mitragyna speciosa varieties. Gels. 2025; 11(7):494.
[35] Haspi NFZ, Mat So'ad SZ, Arzmi MH, Khatib A. Sequential extraction and phytochemical study of the extracts of Mitragyna speciosa. Malays J Chem. 2025; 27(1):84-91.
[36] Kafo AS, Elsalami RM, Hassan M. Mitragyna speciosa Korth. downregulates macrophage inflammatory responses by inhibiting TLR-4 and increasing IL-10 production. İstanbul J Pharm. 2024; 54(3):350-8.
[37] Berthold EC, Kamble SH, Raju KS, King TI, Popa R, Sharma A, et al. Preclinical pharmacokinetic study of speciociliatine, a kratom alkaloid, in rats using an UPLC-MS/MS method. J Pharm Biomed Anal. 2021; 194:113778.
[38] Ramanathan S, Parthasarathy S, Murugaiyah V, Magosso E, Tan SC, Mansor SM. Understanding the physicochemical properties of mitragynine, a principal alkaloid of Mitragyna speciosa, for preclinical evaluation. Molecules. 2015; 20(3):4915-27.
[39] Decosterd I, Woolf CJ. Spared nerve injury: an animal model of persistent peripheral neuropathic pain. Pain. 2000; 87(2):149-58.
[40] Savastano LE, Laurito SR, Fitt MR, Rasmussen JA, Gonzalez Polo V, Patterson SI. Sciatic nerve injury: a simple and subtle model for investigating many aspects of nervous system damage and recovery. J Neurosci Methods. 2014; 227:166-80.
[41] Elsa L, Yuwono M, Prawita A. Pengembangan metode isolasi dan identifikasi mitragynine dalam daun kratom (Mitragyna speciosa). J Biosains Pascasarjana. 2016; 18(3):191.
[42] Indriani DW, Rahmawati SI, Bayu A, Ahmadi P, Sari AN, Zuraida Z, et al. Serotonin release mediates analgesia via opioidergic system and withdrawal symptoms in chronic kratom extract-treated mice. BMC Complement Med Ther. 2025; 25(1):205.
[43] Palit P, Mandal SC, Bhunia B. Total steroid and terpenoid enriched fraction from Euphorbia neriifolia Linn offers protection against nociceptive-pain, inflammation, and in vitro arthritis model: an insight of mechanistic study. Int Immunopharmacol. 2016; 41:106-15.
[44] McNamara CR, Mandel-Brehm J, Bautista DM, Siemens J, Deranian KL, Zhao M, et al. TRPA1 mediates formalin-induced pain. Proc Natl Acad Sci U S A. 2007; 104(33):13525-30.
[45] Razieh Amini Zadeh, Mojtaba Emam Bakhsh, Fariba Sharififar, Rehman NU, Arif M, Esmaeilpour K. Anti-inflammatory and analgesic activity of Hertia intermedia leaf extract in rats. Int J Pharm Integr Health Sci. 2025; 6(1).
[2] Colloca L, Ludman T, Bouhassira D, Baron R, Dickenson AH, Yarnitsky D, et al. Neuropathic pain. Nat Rev Dis Primers. 2017; 3:17002.
[3] Zhou YQ, Liu Z, Liu ZH, Chen SP, Li M, Shahveranov A, et al. Interleukin-6: an emerging regulator of pathological pain. J Neuroinflammation. 2016; 13(1):141.
[4] Lee KM, Jeon SM, Cho HJ. Interleukin-6 induces microglial CX3CR1 expression in the spinal cord after peripheral nerve injury through the activation of p38 MAPK. Eur J Pain. 2010; 14(7):682.e1-12.
[5] Scheller J, Garbers C, Rose-John S. Interleukin-6: from basic biology to selective blockade of pro-inflammatory activities. Semin Immunol. 2014; 26(1):2-12.
[6] Karunakaran T, Ngew KZ, Zailan AAD, Mian Jong VY, Abu Bakar MH. The chemical and pharmacological properties of mitragynine and its diastereomers: an insight review. Front Pharmacol. 2022; 13:805986.
[7] Kruegel AC, Gassaway MM, Kapoor A, Váradi A, Majumdar S, Filizola M, et al. Synthetic and receptor signaling explorations of the Mitragyna alkaloids: mitragynine as an atypical molecular framework for opioid receptor modulators. J Am Chem Soc. 2016; 138(21):6754-64.
[8] Prozialeck WC, Jivan JK, Andurkar SV. Pharmacology of kratom: an emerging botanical agent with stimulant, analgesic and opioid-like effects. J Am Osteopath Assoc. 2012; 112(12):792-9.
[9] Azhar S, Ashari S. Strategies for the development of Mitragyna speciosa (kratom) leaves extract loaded with solid lipid nanoparticles. JMEST. 2019; 6(12):100-4.
[10] Azhar SN, Ashari SE, Tan JK, Kassim NK, Hassan M, Zainuddin N, et al. Screening and selection of formulation components of nanostructured lipid carriers system for Mitragyna speciosa (Korth). Havil drug delivery. Ind Crops Prod. 2023; 198:116668.
[11] Peng Y, Chen L, Ye S, Kang Y, Liu J, Zeng S, et al. Research and development of drug delivery systems based on drug transporter and nano-formulation. Asian J Pharm Sci. 2020; 15(2):220-36.
[12] Dubin AE, Patapoutian A. Nociceptors: the sensors of the pain pathway. J Clin Invest. 2010; 120(11):3760-72.
[13] Cavalli E, Mammana S, Nicoletti F, Bramanti P, Mazzon E. The neuropathic pain: an overview of the current treatment and future therapeutic approaches. Int J Immunopathol Pharmacol. 2019; 33:2058738419838383.
[14] Bouhassira D, Lantéri-Minet M, Attal N, Laurent B, Touboul C. Prevalence of chronic pain with neuropathic characteristics in the general population. Pain. 2008; 136(3):380-7.
[15] van Hecke O, Austin SK, Khan RA, Smith BH, Torrance N. Neuropathic pain in the general population: a systematic review of epidemiological studies. Pain. 2014; 155(9):1907.
[16] Torrance N, Smith BH, Bennett MI, Lee AJ. The epidemiology of chronic pain of predominantly neuropathic origin. Results from a general population survey. J Pain. 2006; 7(4):281-9.
[17] Zimmer Z, Fraser K, Grol-Prokopczyk H, Zajacova A. A global study of pain prevalence across 52 countries: examining the role of country-level contextual factors. Pain. 2022; 163(9):1740-50.
[18] Purwata TE, Sadeli HA, Amir D, Asnawi C, Partoatmodjo L, Aulina S, et al. Characteristics of neuropathic pain in Indonesia: a hospital based national clinical survey. Neurol Asia. 2015; 20(4).
[19] Dureja GP, Iyer RN, Das G, Ahdal J, Narang P. Evidence and consensus recommendations for the pharmacological management of pain in India. J Pain Res. 2017; 10:709-36.
[20] Gilron I, Baron R, Jensen T. Neuropathic pain: principles of diagnosis and treatment. Mayo Clin Proc. 2015; 90(4):532-45.
[21] Wright ME, Rizzolo D. An update on the pharmacologic management and treatment of neuropathic pain. JAAPA. 2017; 30(3):13-7.
[22] Vargas-Schaffer G. Is the WHO analgesic ladder still valid? Twenty-four years of experience. Can Fam Physician. 2010; 56(6):514-7.
[23] Dani M, Guindon J, Lambert C, Beaulieu P. The local antinociceptive effects of paracetamol in neuropathic pain are mediated by cannabinoid receptors. Eur J Pharmacol. 2007; 573(1-3):214-5.
[24] Gulcu N, Karaaslan K, Ozturan KE, Kocoglu H. Intravenous paracetamol for pain relief in a patient with neuropathic pain. Acta Anaesthesiol Scand. 2007; 51(7):956-7.
[25] Mazaleuskaya LL, Sangkuhl K, Thorn CF, FitzGerald GA, Altman RB, Klein TE. PharmGKB summary: pathways of acetaminophen metabolism at the therapeutic versus toxic doses. Pharmacogenet Genomics. 2015; 25(8):416-26.
[26] Ohashi N, Kohno T. Analgesic effect of acetaminophen: a review of known and novel mechanisms of action. Front Pharmacol. 2020; 11:580289.
[27] Im KS, Jung HJ, Kim JB, Lee JM, Park HJ, Joo CH, et al. The antinociceptive effect of acetaminophen in a rat model of neuropathic pain. Kaohsiung J Med Sci. 2012; 28(5):251-8.
[28] Beiranvand S, Sorori MM. Pain management using nanotechnology approaches. Artif Cells Nanomed Biotechnol. 2019; 47(1):462-8.
[29] Gunn J, Hill MM, Cotten BM, Deer TR. An analysis of biomarkers in patients with chronic pain. Pain Physician. 2020; 23(2):235.
[30] Persson AK, Gebauer M, Jordan S, Metz-Weidmann C, Schulte AM, Schneider HC, et al. Correlational analysis for identifying genes whose regulation contributes to chronic neuropathic pain. Mol Pain. 2009; 5:7.
[31] Assanangkornchai S, Muekthong A, Sam-Angsri N, Pattanasattayawong U. The use of Mitragyna speciosa ("krathom"), an addictive plant, in Thailand. Subst Use Misuse. 2007; 42(14):2145-57.
[32] Wahyono S, Widowati L, Handayani L, Sampurno OD, Haryanti S, Fauzi F, et al. Kratom, prospek kesehatan dan sosial ekonomi. Jakarta: Lembaga Penerbit Badan Penelitian dan Pengembangan Kesehatan; 2019.
[33] Stolt AC, Schröder H, Neurath H, Grecksch G, Höllt V, Meyer MR, et al. Behavioral and neurochemical characterization of kratom (Mitragyna speciosa) extract. Psychopharmacology (Berl). 2014; 231(1):13-25.
[34] Anantaworasakul P, Preedalikit W, Anantaworasakul P, Singh S, Intharuksa A, Arunotayanun W, et al. Phytochemical characterization, bioactivities, and nanoparticle-based topical gel formulation development from four Mitragyna speciosa varieties. Gels. 2025; 11(7):494.
[35] Haspi NFZ, Mat So'ad SZ, Arzmi MH, Khatib A. Sequential extraction and phytochemical study of the extracts of Mitragyna speciosa. Malays J Chem. 2025; 27(1):84-91.
[36] Kafo AS, Elsalami RM, Hassan M. Mitragyna speciosa Korth. downregulates macrophage inflammatory responses by inhibiting TLR-4 and increasing IL-10 production. İstanbul J Pharm. 2024; 54(3):350-8.
[37] Berthold EC, Kamble SH, Raju KS, King TI, Popa R, Sharma A, et al. Preclinical pharmacokinetic study of speciociliatine, a kratom alkaloid, in rats using an UPLC-MS/MS method. J Pharm Biomed Anal. 2021; 194:113778.
[38] Ramanathan S, Parthasarathy S, Murugaiyah V, Magosso E, Tan SC, Mansor SM. Understanding the physicochemical properties of mitragynine, a principal alkaloid of Mitragyna speciosa, for preclinical evaluation. Molecules. 2015; 20(3):4915-27.
[39] Decosterd I, Woolf CJ. Spared nerve injury: an animal model of persistent peripheral neuropathic pain. Pain. 2000; 87(2):149-58.
[40] Savastano LE, Laurito SR, Fitt MR, Rasmussen JA, Gonzalez Polo V, Patterson SI. Sciatic nerve injury: a simple and subtle model for investigating many aspects of nervous system damage and recovery. J Neurosci Methods. 2014; 227:166-80.
[41] Elsa L, Yuwono M, Prawita A. Pengembangan metode isolasi dan identifikasi mitragynine dalam daun kratom (Mitragyna speciosa). J Biosains Pascasarjana. 2016; 18(3):191.
[42] Indriani DW, Rahmawati SI, Bayu A, Ahmadi P, Sari AN, Zuraida Z, et al. Serotonin release mediates analgesia via opioidergic system and withdrawal symptoms in chronic kratom extract-treated mice. BMC Complement Med Ther. 2025; 25(1):205.
[43] Palit P, Mandal SC, Bhunia B. Total steroid and terpenoid enriched fraction from Euphorbia neriifolia Linn offers protection against nociceptive-pain, inflammation, and in vitro arthritis model: an insight of mechanistic study. Int Immunopharmacol. 2016; 41:106-15.
[44] McNamara CR, Mandel-Brehm J, Bautista DM, Siemens J, Deranian KL, Zhao M, et al. TRPA1 mediates formalin-induced pain. Proc Natl Acad Sci U S A. 2007; 104(33):13525-30.
[45] Razieh Amini Zadeh, Mojtaba Emam Bakhsh, Fariba Sharififar, Rehman NU, Arif M, Esmaeilpour K. Anti-inflammatory and analgesic activity of Hertia intermedia leaf extract in rats. Int J Pharm Integr Health Sci. 2025; 6(1).
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How to Cite
1.
Noviandomo R, Veterini A, Santosa K, Airlangga P, Waloejo C. Adjunctive Nano-Formulated Kratom (Mytragina Speciosa) with Paracetamol for Neuropathic Pain in Rats: Mechanical Allodynia and Serum Interleukin-6 Outcomes. Arch Anesth & Crit Care. 2026;.
