Berberine hydrochloride improves cognitive deficiency through hippocampal up-regulation of neurotrophins following inhalant self-administration of methamphetamine

Document Type : Original Article


1 Center for Health Related Social and Behavioral Sciences Research, Shahroud University of Medical Sciences, Shahroud, Iran

2 Department of Neuroscience, School of Advanced Technologies in Medicine, Mazandaran University of Medical Sciences, Sari, Iran

3 Student Research Committee, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran

4 Addiction Research Center, Shahroud University of Medical Sciences, Shahroud, Iran

5 Department of Addiction Studies, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran


Objective(s): Chronic methamphetamine (METH) abuse is recognized as an important risk factor for cognitive impairment. A plant-based isoquinoline alkaloid, Berberine hydrochloride (BER), shows memory and cognition enhancement properties. Due to the aim of the present study which is to investigate the influence of BER administration on METH-induced cognitive deficits, we investigated neurotrophin signaling including brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) as a possible mechanism by which BER exerts its cognitive improvement influences. 
Materials and Methods: In this experimental study, thirty-two male Wistar rats were randomly classified into four groups, including non-treated control, intubated control, METH-inhaled, and METH-inhaled + BER-intubated. Rats in the METH-inhaled group underwent METH inhalation for 14 days, and the BER-inhaled and BER-intubated rats were intubated (100mg/kg) for the following three weeks. A novel object recognition task (NORt) was carried out on days 36 and 37. Rats were sacrificed for histological preparations after the behavioral tests. Neurotrophic factors, including GDNF and BDNF, were evaluated by immunofluorescence staining in the hippocampus. 
Results: This experiment indicated a dramatic improvement in cognitive deficits associated with chronic METH abuse (P<0.001). Furthermore, a significant decrease in the expression of both neurotrophins, GDNF (P<0.001) and BDNF (P<0.001), was observed in the METH-inhaled group compared with the METH-inhaled group treated with BER and non-treated control group. 
Conclusion: Activation of neurotrophic factors after BER administration resulted in improvement of METH-induced cognitive deficits. Therefore, BER may be considered a promising treatment for METH users who experience cognition deficits.


1. Canton H. United Nations Office on Drugs and Crime—UNODC.  The Europa Directory of International Organizations 2021: Routledge; 2021. p. 240-244.
2. Lieb R, Schuetz CG, Pfister H, Von Sydow K, Wittchen H-U. Mental disorders in ecstasy users: a prospective-longitudinal investigation. Drug Alcohol Depend 2002; 68:195-207.
3. L. Simon CD, Jennifer Carnell, Paul Brethen Brethen, Richard Rawson, Walter Ling, Sara. Cognitive impairment in individuals currently using methamphetamine. Am J Addict 2000; 9:222-231.
4. Kalechstein AD, Newton TF, Green M. Methamphetamine dependence is associated with neurocognitive impairment in the initial phases of abstinence. J Neuropsychiatry Clin Neurosci 2003; 15:215-220.
5. Chung A, Lyoo IK, Kim SJ, Hwang J, Bae SC, Sung YH, et al. Decreased frontal white-matter integrity in abstinent methamphetamine abusers. Int J Neuropsychopharmacol 2007; 10:765-775.
6. Ornstein TJ, Iddon JL, Baldacchino AM, Sahakian BJ, London M, Everitt BJ, et al. Profiles of cognitive dysfunction in chronic amphetamine and heroin abusers. Neuropsychopharmacol 2000; 23:113-126.
7. Ersche KD, Clark L, London M, Robbins TW, Sahakian BJ. Profile of executive and memory function associated with amphetamine and opiate dependence. Neuropsychopharmacol 2006; 31:1036-1047.
8. Reske M, Eidt CA, Delis DC, Paulus MP. Nondependent stimulant users of cocaine and prescription amphetamines show verbal learning and memory deficits. Biol Psychiatry 2010; 68:762-769.
9. Osman M, Asiri RA, Almalki SE, Qassadi AM, Alotaibi FS, AlJemaiah A. Screening for Cognitive Dysfunction in Amphetamine Users in Saudi Arabia; a Case-control Investigation Using Propensity Score Matching Analysis. J Psychoactive Drugs 2021:1-8.
10. Bisagno V, Ferguson D, Luine VN. Short toxic methamphetamine schedule impairs object recognition task in male rats. Brain Res 2002; 940:95-101.
11. Schröder N, O’Dell SJ, Marshall JF. Neurotoxic methamphetamine regimen severely impairs recognition memory in rats. Synapse 2003; 49:89-96.
12. Belcher AM, O’Dell SJ, Marshall JF. Impaired Object Recognition Memory Following Methamphetamine, but not p-Chloroamphetamine- or d-Amphetamine-Induced Neurotoxicity. Neuropsychopharmacol 2005; 30:2026-2034.
13. Green AR, Mechan AO, Elliott JM, O’Shea E, Colado MI. The pharmacology and clinical pharmacology of 3, 4-methylenedioxymethamphetamine (MDMA,“ecstasy”). Pharmacol Rev 2003; 55:463-508.
14. Clark RE, Zola SM, Squire LR. Impaired recognition memory in rats after damage to the hippocampus. J Neurosci 2000; 20:8853-8860.
15. de Lima MNM, Presti-Torres J, Vedana G, Alcalde LA, Stertz L, Fries GR, et al. Early life stress decreases hippocampal BDNF content and exacerbates recognition memory deficits induced by repeated D-amphetamine exposure. Behav Brain Res 2011; 224:100-106.
16. Fries GR, Valvassori SS, Bock H, Stertz L, da Silva Magalhaes PV, Mariot E, et al. Memory and brain-derived neurotrophic factor after subchronic or chronic amphetamine treatment in an animal model of mania. J Psychiatr Res 2015; 68:329-336.
17. Grimm JW, Lu L, Hayashi T, Hope BT, Su T-P, Shaham Y. Time-dependent increases in brain-derived neurotrophic factor protein levels within the mesolimbic dopamine system after withdrawal from cocaine: implications for incubation of cocaine craving. J Neurosci 2003; 23:742-747.
18. Angelucci F, Gruber SH, El Khoury A, Tonali PA, Mathé AA. Chronic amphetamine treatment reduces NGF and BDNF in the rat brain. Eur Neuropsychopharmacol 2007; 17:756-762.
19. Ghitza UE, Zhai H, Wu P, Airavaara M, Shaham Y, Lu L. Role of BDNF and GDNF in drug reward and relapse: a review. Neurosci Biobehav Rev 2010; 35:157-171.
20. Chen PH, Huang MC, Lai YC, Chen PY, Liu HC. Serum brain‐derived neurotrophic factor levels were reduced during methamphetamine early withdrawal. Addict Biol 2014; 19:482-485.
21. Ciketic S, Hayatbakhsh MR, Doran CM, Najman JM, McKetin R. A review of psychological and pharmacological treatment options for methamphetamine dependence. J Subst Use 2012; 17:363-383.
22. Karila L, Weinstein A, Aubin HJ, Benyamina A, Reynaud M, Batki SL. Pharmacological approaches to methamphetamine dependence: a focused review. Br J Clin Pharmacol 2010; 69:578-592.
23. Imenshahidi M, Hosseinzadeh H. Berberis vulgaris and berberine: an update review. Phytother Res 2016; 30:1745-1764.
24. Shen J-d, Ma L-g, Hu C-y, Pei Y-y, Jin S-l, Fang X-y, et al. Berberine up-regulates the BDNF expression in hippocampus and attenuates corticosterone-induced depressive-like behavior in mice. Neurosci Lett
 2016; 614:77-82.
25. Yang J, Yan H, Li S, Zhang M. Berberine ameliorates MCAO induced cerebral ischemia/reperfusion injury via activation of the BDNF–TrkB–PI3K/Akt signaling pathway. Neurochem Res 2018; 43:702-710.
26. Rashtbari H, Razi M, Hassani-Bafrani H, Najaran H. Berberine reinforces Sertoli cells niche and accelerates spermatogonial stem cells renewal in experimentally-induced varicocele condition in rats. Phytomedicine 2018; 40:68-78.
27. Rafaiee R, Ahmadiankia N, Mousavi SA, Rezaeian L, Niroumand Sarvandani M, Shekari A, et al. Inhalant self-administration of methamphetamine: the most similar model to human methamphetamine addiction. Ir J Psychiatr Behav Sci 2019; 13.
28. Moghaddam HK, Baluchnejadmojarad T, Roghani M, Goshadrou F, Ronaghi A. Berberine chloride improved synaptic plasticity in STZ induced diabetic rats. Metab Brain Dis 2013; 28:421-428.
29. Antunes M, Biala G. The novel object recognition memory: neurobiology, test procedure, and its modifications. Cogn process 2012; 13:93-110.
30. Mohseni F, Khaksari M, Rafaiee R, Rahimi K, Norouzi P, Garmabi B. Apelin 13 improves anxiety and cognition via hippocampal increases BDNF expression and reduction cell death in neonatal alcohol exposed rats. Int J Pept Res Ther 2021; 27:1351-1362.
31. Mizoguchi H, Yamada K. Methamphetamine use causes cognitive impairment and altered decision-making. Neurochem Int 2019; 124:106-113.
32. Ravandi SG, Shabani M, Bashiri H, Goraghani MS, Khodamoradi M, Nozari M. Ameliorating effects of berberine on MK-801-induced cognitive and motor impairments in a neonatal rat model of schizophrenia. Neurosci Lett 2019; 706:151-157.
33. Singh A, Dhaneshwar S, Mazumder A. Investigating Neuroprotective Potential of Berberine, Levetiracetam and Their Combination in The Management of Alzheimer’s Disease Utilizing Drug Repurposing Strategy. Curr Rev Clin Exp Pharmacol 2021.
34. Yuan N-N, Cai C-Z, Wu M-Y, Su H-X, Li M, Lu J-H. Neuroprotective effects of berberine in animal models of Alzheimer’s disease: A systematic review of pre-clinical studies. BMC Complement Alternat med 2019; 19:1-10.
35. Hasanitabar A, Fatholahi M, Bitaraf A, Mahdi S. Evaluation of Anti-Inflammatory Effect of Combination Therapy of Silymarin Nanomicelles plus Berberine Nanomicelles in LPS-induced Depressive-like Behavior in Mice. Acta Sci Pharm 2021; 2.
36. Fan J, Zhang K, Jin Y, Li B, Gao S, Zhu J, et al. Pharmacological effects of berberine on mood disorders. J Cell Mol Med 2019; 23:21-28.
37. Fang Y, Zhang J, Zhu S, He M, Ma S, Jia Q, et al. Berberine ameliorates ovariectomy-induced anxiety-like behaviors by enrichment in equol generating gut microbiota. Pharmacol Res 2021; 165:105439.
38. Zhu J-r, Lu H-d, Guo C, Fang W-r, Zhao H-d, Zhou J-s, et al. Berberine attenuates ischemia–reperfusion injury through inhibiting HMGB1 release and NF-κB nuclear translocation. Acta Pharmacol Sin 2018; 39:1706-1715.
39. Dadgostar E, Moghanlou M, Parvaresh M, Mohammadi S, Khandan M, Aschner M, et al. Can Berberine Serve as a New Therapy for Parkinson’s Disease? Neurotoxicity Research 2022:1-7.
40. Rezaeian L, Kalalian-Moghaddam H, Mohseni F, Khaksari M, Rafaiee R. Effects of berberine hydrochloride on methamphetamine-induced anxiety behaviors and relapse in rats. Iran J Basic Med Sci 2020; 23:1480.
41. Alavijeh MM, Vaezi G, Khaksari M, Hojati V. Berberine hydrochloride attenuates voluntary methamphetamine consumption and anxiety-like behaviors via modulation of oxytocin receptors in methamphetamine addicted rats. Physiol Behav 2019; 206:157-165.
42. Rezaeian L, Khaksari M, Rafaiee R, Kalalian Moghaddam H. Neuroprotective Effects of Berberine Hydrochloride on Methamphetamine-induced Cognitive Dysfunction: Immunohistochemical and Behavioral Studies in Rats. Basic Clin Neurosci J 2022; 13:443-454.
43. Mohseni F, Behnam SG, Rafaiee R. A Review of the historical evolutionary process of dry and water maze tests in rodents. Basic Clin Neurosci 2020; 11:389-402.
44. Bevins RA, Besheer J. Object recognition in rats and mice: a one-trial non-matching-to-sample learning task to study’recognition memory’. Nat Protoc 2006; 1:1306-1311.
45. Brown MW, Aggleton JP. Recognition memory: what are the roles of the perirhinal cortex and hippocampus? Nat Rev Neurosci 2001; 2:51-61.
46. Hernández‐Hernández EM, Caporal Hernandez K, Vázquez‐Roque RA, Díaz A, de la Cruz F, Florán B, et al. The neuropeptide‐12 improves recognition memory and neuronal plasticity of the limbic system in old rats. Synapse 2018; 72:e22036.
47. Chaves Filho AJM, Cunha NL, de Araújo Rodrigues P, de Souza AG, Soares MV-R, Jucá PM, et al. Doxycycline reverses cognitive impairment, neuroinflammation and oxidative imbalance induced by D-amphetamine mania model in mice: A promising drug repurposing for bipolar disorder treatment? European Neuropsychopharmacol 2021; 42:57-74.
48. Marshall JF, Belcher AM, Feinstein EM, O’Dell SJ. Methamphetamine‐induced neural and cognitive changes in rodents. Addiction 2007; 102:61-69.
49. Belcher AM, O’Dell SJ, Marshall JF. Impaired object recognition memory following methamphetamine, but not p-chloroamphetamine-or d-amphetamine-induced neurotoxicity. Neuropsychopharmacol 2005; 30:2026-2034.
50. Shi J, Xu H, Cavagnaro MJ, Li X, Fang J. Blocking HMGB1/RAGE Signaling by Berberine Alleviates A1 Astrocyte and Attenuates Sepsis-Associated Encephalopathy. Front Pharmacol 2021:777-780.
51. Kahale V, Mhaiskar A, Shelat P, Pooja R, Gaikwad N, Mundhada D. To determine the Effect of Berberine on 6-OHDA induced memory impairment in Parkinson’s disease in rodents. J Pharm Innov 2014; 3:101.
52. Teixeira AL, Barbosa IG, Diniz BS, Kummer A. Circulating levels of brain-derived neurotrophic factor: correlation with mood, cognition and motor function. Biomark Med 2010; 4:871-887.
53. Gunstad J, Benitez A, Smith J, Glickman E, Spitznagel MB, Alexander T, et al. Serum brain-derived neurotrophic factor is associated with cognitive function in healthy older adults. J Geriatr Psychiatry Neurol 2008; 21:166-170.
54. Pelleymounter MA, Cullen MJ, Baker MB, Healy D. Glial cell line-derived neurotrophic factor (GDNF) improves spatial learning in aged Fischer 344 rats. Psychobiol 1999; 27:397-401.
55. Pertusa M, Garcia-Matas S, Mammeri H, Adell A, Rodrigo T, Mallet J, et al. Expression of GDNF transgene in astrocytes improves cognitive deficits in aged rats. Neurobiol Aging 2008; 29:1366-1379.
56. Yamada K, Nabeshima T. Brain-derived neurotrophic factor/TrkB signaling in memory processes. J Pharmacol Sci 2003; 91:267-270.
57. Furini CR, Rossato JI, Bitencourt LL, Medina JH, Izquierdo I, Cammarota M. β‐Adrenergic receptors link NO/sGC/PKG signaling to BDNF expression during the consolidation of object recognition long‐term memory. Hippocampus 2010; 20:672-683.
58.Brasnjevic I, Steinbusch HW, Schmitz C, Martinez-Martinez P, Initiative ENR. Delivery of peptide and protein drugs over the blood–brain barrier. Prog Neurobiol 2009; 87:212-251.
59. Kastin AJ, Akerstrom V, Pan W. Glial cell line-derived neurotrophic factor does not enter normal mouse brain. Neurosci Lett 2003; 340:239-241.
60. Pardridge WM. Delivery of biologics across the blood–brain barrier with molecular Trojan horse technology. BioDrugs 2017; 31:503-519.
61. Singh S, Ahmad R, Mathur D, Sagar RK, Krishana B, Arora R, et al. Neuroprotective effect of BDNF in young and aged 6-OHDA treated rat model of Parkinson disease.  2006; 44:699-704.
62. Wang L, Sheng W, Tan Z, Ren Q, Wang R, Stoika R, et al. Treatment of Parkinson’s disease in Zebrafish model with a berberine derivative capable of crossing blood brain barrier, targeting mitochondria, and convenient for bioimaging experiments. Comparative Biochemistry and Physiology Part C: Toxicol  Pharmacol 2021; 249:109151.
63. Chen S-Y, Gao Y, Sun J-Y, Meng X-L, Yang D, Fan L-H, et al. Traditional Chinese medicine: role in reducing β-amyloid, apoptosis, autophagy, neuroinflammation, oxidative stress, and mitochondrial dysfunction of Alzheimer’s disease. Front pharmacol 2020; 11:497.
64. Shakouri E, Azarbayjani MA, Jameie SB, Peeri M, Farhadi M. Berberine supplement and resistance training may ameliorate diazinon induced neural toxicity in rat hippocampus via the activation of the TrkB and ERK signaling pathway. Int Clin Neurosci J 2021; 8:14-21.
65. Liu P, Li Y, Qi X, Xu J, Liu D, Ji X, et al. Protein kinase C is involved in the neuroprotective effect of berberine against intrastriatal injection of quinolinic acid‐induced biochemical alteration in mice. J Cell Mol Med 2019; 23:6343-6354.
66.Oliveira JSd. Efeito neuroprotetor da Berberina na neurotoxicidade induzida por estreptozotocina e lipolissacarídeo em ratos [Doctoral thesis]: Universidade Federal de Santa Maria; 2019.