EVALUATING THE THERAPEUTIC EFFICACY OF METFORMIN-BERBERINE LOADED NANOPARTICLES FOR THE TREATMENT OF TYPE 2 DIABETES INDUCED ALZHEIMER’S DISEASE

RAVINA YADAV; RUCHI JAKHMOLA MANI; ARUN K. SHARMA; ASHISH KUMAR; DEEPSHIKHA PANDE KATARE

doi:10.22159/ijap.2026v18i1.56212

Authors

RAVINA YADAV Proteomics and Translational Research Lab, Centre for Medical Biotechnology, Amity Institute of Biotechnology, Amity University, Noida-201301, India https://orcid.org/0000-0001-6841-6166
RUCHI JAKHMOLA MANI Proteomics and Translational Research Lab, Centre for Medical Biotechnology, Amity Institute of Biotechnology, Amity University, Noida-201301, India
ARUN K. SHARMA Department of Pharmacology, Amity Institute of Pharmacy, Amity University, Gurugram-122413, Haryana, India https://orcid.org/0000-0002-5692-6502
ASHISH KUMAR Department of Pharmaceutics, Amity Institute of Pharmacy, Amity University, Gurugram-122413, Haryana, India
DEEPSHIKHA PANDE KATARE Department of Pharmaceutics, Amity Institute of Pharmacy, Amity University, Gurugram-122413, Haryana, India

DOI:

https://doi.org/10.22159/ijap.2026v18i1.56212

Keywords:

Type 2 diabetes mellitus, Alzheimer’s disease, Liver-brain axis, Berberine hydrochloride, Solid lipid nanoparticles, Targeted drug delivery

Abstract

Objective: To develop and evaluate a novel drug delivery system (NDDS) of Metformin and Berberine- loaded solid lipid nanoparticles (MBSLNPs) for neuroprotection in type 2 diabetes mellitus (T2DM) induced Alzheimer’s disease (AD) model. Methods: MBSLNPs were prepared by hot homogenization and ultrasonication, characterized for particle size, zeta potential, drug loading, and FTIR (fourier transform infrared spectroscopy) compatibility. In vitro release kinetics were modeled, and in vivo efficacy was assessed in a combined streptozotocin (STZ) and amyloid- β (Aβ) rat model by performing behavioral, biochemical, and histopathological analyses. Safety of blank SLNs was evaluated in healthy rats. Results: MBSLNPs exhibited a mean particle size of 102±5nm, zeta potential of –29±3mV, and high drug loading, with no FTIR incompatibilities. Drug release followed first-order kinetics (R²=0.9955) with Fickian diffusion (n=0.43). In Vivo, MBSLNPs significantly improved memory performance in Morris Water Maze and Y-Maze compared with free drug (p<0.01) and disease control (p<0.001). Oxidative stress markers like malondialdehyde (MDA), thiobarbituric acid reactive substances (TBARS) were reduced, while antioxidant enzymes like superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione (GSH) increased significantly (p<0.01 vs. disease control). Pro-inflammatory cytokines (TNF-α, IL-6, NF-κB) decreased markedly (p<0.001), and acetylcholinesterase (AChE) activity was normalized (p<0.01 vs. free drug) in addition to lower cortical and hippocampal Aβ-40/42 levels in MBSLNP-treated rats (p<0.001). Semi-quantitative histopathology showed significantly reduced neuronal loss and gliosis compared with both disease and free drug groups. Safety evaluation of blank SLNs revealed no significant changes in Serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), creatinine, or blood nitrogen (BUN). Conclusion: MBSLNPs demonstrate superior neuroprotective and hepatoprotective efficacy over free drugs, suggesting their translational potential as a combined therapeutic strategy for AD with concurrent liver dysfunction. Thiobarbituric acid reactive substances (TBARS), glutathione peroxidase (GPx), acetylcholinesterase (AChE), serum glutamate oxaloacetate transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT) [26], blood glucose levels, and histological analysis of brain tissue.

References

1. Bacsu JD, Mah JC, Jamali AA, Conanan C, Lautrup S, Berger C, Fiske D, Fraser S, Virani A, Fehr F, Chasteen AL. Stigma of Dementia on Social Media During World Alzheimer’s Awareness Month: Thematic Analysis of Posts. JMIR Formative Research. 2025 Jun 2;9(1):e72775.

2. Kciuk M, Kruczkowska W, Gałęziewska J, Wanke K, Kałuzińska-Kołat Ż, Aleksandrowicz M, Kontek R. Alzheimer’s disease as type 3 diabetes: Understanding the link and implications. International Journal of Molecular Sciences. 2024 Nov 7;25(22):11955.

3. Meng X, Zhang H, Zhao Z, Li S, Zhang X, Guo R, Liu H, Yuan Y, Li W, Song Q, Liu

J. Type 3 diabetes and metabolic reprogramming of brain neurons: causes and therapeutic strategies. Molecular Medicine. 2025 Feb 18;31(1):61.

4. Li X, Song D, Leng SX. Link between type 2 diabetes and Alzheimer’s disease: from epidemiology to mechanism and treatment. Clinical interventions in aging. 2015 Mar 10:549-60.

5. Cheon SY, Song J. Novel insights into non-alcoholic fatty liver disease and dementia: insulin resistance, hyperammonemia, gut dysbiosis, vascular impairment, and inflammation. Cell & Bioscience. 2022 Jun 28;12(1):99.

6. Bosoi CR, Vandal M, Tournissac M, Leclerc M, Fanet H, Mitchell PL, Verreault M, Trottier J, Virgili J, Tremblay C, Lippman HR. High‐Fat Diet Modulates Hepatic Amyloid β and Cerebrosterol Metabolism in the Triple Transgenic Mouse Model of Alzheimer’s Disease. Hepatology Communications. 2021 Mar;5(3):446-60.

7. Rani A, Tabassum F, Sharma AK. Molecular interplay between phytoconstituents of Ficus Racemosa and neurodegenerative diseases. European Journal of Neuroscience. 2024 Apr;59(7):1833-47.

8. Nishida Y. Labile Iron Controllng Therapy To Prevent Cancers and Alzheimer’s Disease. Int J Chem. 2025 Aug 7;17(2):77.

9. Onyango IG, Jauregui GV, Čarná M, Bennett Jr JP, Stokin GB. Neuroinflammation in Alzheimer’s disease. Biomedicines. 2021 May 7;9(5):524.

10. Adamu A, Li S, Gao F, Xue G. The role of neuroinflammation in neurodegenerative diseases: current understanding and future therapeutic targets. Frontiers in aging neuroscience. 2024 Apr 12;16:1347987.

11. Sharma AK, Taneja G, Khanna D, Rajput SK. Reactive oxygen species: friend or foe?. RSC Advances. 2015;5(71):57267-76.

12. Grossberg GT. Cholinesterase inhibitors for the treatment of Alzheimer's disease:: getting on and staying on. Current Therapeutic Research. 2003 Apr 1;64(4):216-35.

13. Aderinto N, Olatunji G, Abdulbasit M, Ashinze P, Faturoti O, Ajagbe A, Ukoaka B, Aboderin G. The impact of diabetes in cognitive impairment: A review of current evidence and prospects for future investigations. Medicine. 2023 Oct 27;102(43):e35557.

14. Bian X, Guo Q, Yau LF, Yang L, Wang X, Zhao S, Wu S, Qin X, Jiang ZH, Li C. Berberine-inspired ionizable lipid for self-structure stabilization and brain targeting

delivery of nucleic acid therapeutics. Nature Communications. 2025 Mar 10;16(1):2368.

15. Li Z, Geng YN, Jiang JD, Kong WJ. Antioxidant and anti‐inflammatory activities of berberine in the treatment of diabetes mellitus. Evidence‐Based Complementary and Alternative Medicine. 2014;2014(1):289264.

16. Li N, Zhou T, Fei E. Actions of metformin in the brain: a new perspective of metformin treatments in related neurological disorders. International Journal of Molecular Sciences. 2022 Jul 27;23(15):8281.

17. Satapathy MK, Yen TL, Jan JS, Tang RD, Wang JY, Taliyan R, Yang CH. Solid lipid nanoparticles (SLNs): an advanced drug delivery system targeting brain through BBB. Pharmaceutics. 2021 Jul 31;13(8):1183.

18. Fakhri S, Abdian S, Zarneshan SN, Moradi SZ, Farzaei MH, Abdollahi M. Nanoparticles in combating neuronal dysregulated signaling pathways: recent approaches to the nanoformulations of phytochemicals and synthetic drugs against neurodegenerative diseases. International journal of nanomedicine. 2022 Jan 1:299- 331.

19. Jakhmola Mani R, Dogra N, Katare DP. The connection between chronic liver damage and sporadic Alzheimer’s disease: evidence and insights from a rat model. Brain Sciences. 2023 Sep 29;13(10):1391.

20. Sharma AK, Kumar A, Kumar S, Mukherjee S, Nagpal D, Nagaich U, Rajput SK. Preparation and therapeutic evolution of Ficus benjamina solid lipid nanoparticles against alcohol abuse/antabuse induced hepatotoxicity and cardio-renal injury. RSC advances. 2017;7(57):35938-49.

21. Magdum SV, Shirote PJ. Formulation and Evaluation of Candesartan Cilexetil Nanosuspension for Oral Drug Delivery System. Int. J. Pharm. Sci. Drug Res., March

- April 2025, Vol 17, Issue 2, 159-168.

22. Bhatt F, Patel D. Fabrication and screening of solid lipid nanoparticles-loaded microneedle patch for polycystic ovary syndrome treatment. Asian Journal of Pharmaceutical and Clinical Research .2025.18(7):109–114.

23. Deep A, Verma M, Marwaha RK, Sharma AK, Kumari B. Development, characterization and anticancer evaluation of silver nanoparticles from Dalbergia sissoo leaf extracts. Current Cancer Therapy Reviews. 2020 Jun 1;16(2):145-51.

24. Shallie OF, Mabandla MV. Amyloid-beta (1-42) lesion of CA1 rat dorsal hippocampus reduces contextual fear memory and increases expression of microglial genes regulating neuroinflammation. Behavioural Brain Research. 2020 Sep 1;393:112795.

25. Xuan WT, Wang H, Zhou P, Ye T, Gao HW, Ye S, Wang JH, Chen ML, Song H, Wang Y, Cai B. Berberine ameliorates rats model of combined Alzheimer’s disease and type 2 diabetes mellitus via the suppression of endoplasmic reticulum stress. 3 Biotech. 2020 Aug;10(8):359.

26. Kokani H, Sawant P, Sonawane P, et al. Herbal therapies for liver diseases: An overview. International Journal of Pharmacy and Pharmaceutical Science . 2025 [cited 2025 Aug 16;7(1):140–144.

27. Wang L, Liu W, Fan Y, Liu T, Yu C. Effect of rosiglitazone on amyloid precursor protein processing and Aβ clearance in streptozotocin-induced rat model of Alzheimer’s disease. Iranian journal of basic medical sciences. 2017 May;20(5):474.

28. Hoffman JL, Faccidomo S, Kim M, Taylor SM, Agoglia AE, May AM, Smith EN, Wong LC, Hodge CW. Alcohol drinking exacerbates neural and behavioral pathology in the 3xTg-AD mouse model of Alzheimer's disease. International review of neurobiology. 2019 Jan 1;148:169-230.

29. Çınar E, Mutluay SU, Baysal İ, Gültekinoğlu M, Ulubayram K, Çiftçi SY, Tel BC, Uçar G. Donepezil-loaded PLGA-b-PEG nanoparticles enhance the learning and memory function of beta-amyloid rat model of Alzheimer’s disease. Archives of Neuropsychiatry. 2022 Nov 19;59(4):281.

30. Kumari N, Mittal A, Rana A, Sharma AK. Callistemon viminalis extracts: dual action on anxiety and neuroprotection via neuroinflammatory and serotonergic pathways. Neuroscience and Behavioral Physiology. 2025 Jan;55(1):61-73.

31. Sharma V, Singh J, Kumar Y, Kumar A, Venkatesan K, Mukherjee M, Sharma AK. Integrated insights into gene expression dynamics and transcription factor roles in diabetic and diabetic-infectious wound healing using rat model. Life Sciences. 2025 May 1;368:123508.

32. VV L. Formulation and evaluation of polyherbal cream. International Journal of Pharmacy and Pharmaceutical Science . 2025 Aug 16;7(2):36–42.

33. Corti G, Capasso G, Maestrelli F, Cirri M, Mura P. Physical–chemical characterization of binary systems of metformin hydrochloride with triacetyl-β- cyclodextrin. Journal of pharmaceutical and biomedical analysis. 2007 Nov 5;45(3):480-6.

34. Battu SK, Repka MA, Maddineni S, Chittiboyina AG, Avery MA, Majumdar S. Physicochemical characterization of berberine chloride: a perspective in the development of a solution dosage form for oral delivery. Aaps Pharmscitech. 2010 Sep;11(3):1466-75.

35. Cheng CL, Lawrence XY, Lee HL, Yang CY, Lue CS, Chou CH. Biowaiver extension potential to BCS Class III high solubility-low permeability drugs: bridging evidence for metformin immediate-release tablet. European Journal of Pharmaceutical Sciences. 2004 Jul 1;22(4):297-304.

36. Solnier J, Zhang Y, Kuo YC, Du M, Roh K, Gahler R, Wood S, Chang C. Characterization and pharmacokinetic assessment of a new berberine formulation with enhanced absorption in vitro and in human volunteers. Pharmaceutics. 2023 Nov 1;15(11):2567.

37. Metry M, Shu Y, Abrahamsson B, Cristofoletti R, Dressman JB, Groot DW, Parr A, Langguth P, Shah VP, Tajiri T, Mehta MU. Biowaiver monographs for immediate release solid oral dosage forms: metformin hydrochloride. Journal of Pharmaceutical Sciences. 2021 Apr 1;110(4):1513-26.

38. Narade S, Pore Y. Optimization of ex vivo permeability characteristics of berberine in presence of quercetin using 32 full factorial design. Journal of Applied Pharmaceutical Science. 2019 Feb 4;9(1):073-82.

39. Kurul F, Turkmen H, Cetin AE, Topkaya SN. Nanomedicine: How nanomaterials are transforming drug delivery, bio-imaging, and diagnosis. Next Nanotechnology. 2025 Jan 1;7:100129.

40. Rojek B, Wesolowski M. Fourier transform infrared spectroscopy supported by multivariate statistics in compatibility study of atenolol with excipients. Vibrational Spectroscopy. 2016 Sep 1;86:190-7.

41. Thuan ND, Cuong HM, Nam NH, Huong NT, Hong HS. Morphological analysis of Pd/C nanoparticles using SEM imaging and advanced deep learning. RSC advances. 2024;14(47):35172-83.

42. George IE, Cherian T, Ragavendran C, Mohanraju R, Dailah HG, Hassani R, Alhazmi HA, Khalid A, Mohan S. One-pot green synthesis of silver nanoparticles using brittle star Ophiocoma scolopendrina: Assessing biological potentialities of antibacterial, antioxidant, anti-diabetic and catalytic degradation of organic dyes. Heliyon. 2023 Mar 1;9(3).

43. de la Monte SM, Longato L, Tong M, Wands JR. Insulin resistance and neurodegeneration: roles of obesity, type 2 diabetes mellitus and non-alcoholic steatohepatitis. Current opinion in investigational drugs (London, England: 2000). 2009 Oct;10(10):1049.

44. Gomes BA, Santos SM, Gato LD, Espíndola KM, Silva RK, Davis K, Navegantes- Lima KC, Burbano RM, Romao PR, Coleman MD, Monteiro MC. Alpha-lipoic acid reduces neuroinflammation and oxidative stress induced by dapsone in an animal model. Nutrients. 2025 Feb 25;17(5):791.

45. Chen ZR, Huang JB, Yang SL, Hong FF. Role of cholinergic signaling in Alzheimer’s disease. Molecules. 2022 Mar 10;27(6):1816.

46. Abd Elmonem HA, Morsi RM, Mansour DS, El-Sayed ES. Myco-fabricated ZnO nanoparticles ameliorate neurotoxicity in mice model of Alzheimer’s disease via acetylcholinesterase inhibition and oxidative stress reduction. Biometals. 2023 Dec;36(6):1391-404.

47. Singhal A, Morris VB, Labhasetwar V, Ghorpade A. Nanoparticle-mediated catalase delivery protects human neurons from oxidative stress. Cell death & disease. 2013 Nov;4(11):e903-.

48. Kodi T, Praveen S, Paka SK, Sankhe R, Gopinathan A, Krishnadas N, Kishore A. Neuroprotective Effects of Metformin and Berberine in Lipopolysaccharide‐Induced Sickness‐Like Behaviour in Mice. Advances in Pharmacological and Pharmaceutical Sciences. 2024;2024(1):8599268.