Combination of Insulin and UC-MSCs Secretome Therapy Demonstrates A Trend of Liver Injury Improvement in Diabetic Metabolic Dysfunction-associated Steatotic Liver Disease (MASLD) Rat Model
Abstract
BACKGROUND: Metabolic dysfunction-associated steatotic liver disease (MASLD) coexists with diabetes mellitus and presents a significant therapeutic challenge due to its complex metabolic and hepatic pathology. Insulin therapy remains the cornerstone for glycemic management, but its effects on hepatic steatosis is inconsistent. Complementary therapies are needed to address this limitation. Umbilical cord-derived mesenchymal stem cells (UC-MSCs) secretome has emerged as therapeutic approach for metabolic disease due to its anti-inflammatory, immunomodulatory, and antioxidant properties. This study was conducted to evaluate whether adjunctive administration of UC-MSCs secretome could enhance therapeutic outcomes, particularly in alleviating hepatic steatosis in a diabetic MASLD rat model.
METHODS: Male rats were grouped into normal and diabetic MASLD rat model, and treated with either growth medium, insulin, UC-MSCs secretome, or combination of insulin and UC-MSCs secretome for 4 weeks. Blood samples were collected and liver enzymatic and lipid profile were analyzed using chemistry analyzer. Liver tissue was also obtained for assessment of liver fibrosis using Hematoxylin-Eosin staining, interleukin (IL)-6 measurement using enzyme-linked immunosorbent assay (ELISA), and superoxide dismutase (SOD) activity measurement using colorimetry.
RESULTS: Combined insulin and UC-MSCs secretome therapy showed a trend toward better blood glucose management and lipid parameters, although not statistically significant. Administration of insulin, secretome, and their combination was associated with higher hepatic SOD levels and lower IL-6 protein levels, although not statistically significant. Combined therapy also reduced hepatomegaly compared to untreated diabetic MASLD rat model, and improved hepatic steatosis while significantly alleviating fibrosis severity.
CONCLUSION: Combination of insulin and UC-MSCs secretome therapy in diabetic MASLD was able to improve several parameters related to hepatic steatosis compared with the untreated diabetic MASLD rat model. These findings suggest that UC-MSCs secretome might serve as a potential adjunct to insulin therapy.
KEYWORDS: metabolic dysfunction-associated steatotic liver disease, type 2 diabetes, cell free therapy, umbilical cord mesenchymal stem cells, secretome
References
International Diabetes Federation (IDF). IDF Diabetes Atlas 11th Edition. 11th ed. Brussels: International Diabetes Federation; 2025.
Wiafe YA, Afihene MY, Anto EO, Nmai RA, Amoah-Kumi L, Frimpong J, et al. Non-alcoholic fatty liver disease and liver fibrosis in persons with type 2 diabetes mellitus in Ghana: A study of prevalence, severity, and contributing factors using transient elastography. J Clin Med. 2023; 12(11): 3741, CrossRef.
Ajmera V, Cepin S, Tesfai K, Hofflich H, Cadman K, Lopez S, et al. A prospective study on the prevalence of NAFLD, advanced fibrosis, cirrhosis and hepatocellular carcinoma in people with type 2 diabetes. J Hepatol. 2023; 78(3): 471-8, CrossRef.
Younossi ZM, Golabi P, de Avila L, Paik JM, Srishord M, Fukui N, et al. The global epidemiology of NAFLD and NASH in patients with type 2 diabetes: A systematic review and meta-analysis. J Hepatol. 2019; 71(4): 793-801, CrossRef.
Sulaeman A, Amiruddin AR, Lawrence GS. Metabolic syndrome (MetS) and nonalcoholic steatohepatitis (NASH): Study of biochemical markers free fatty acid (FFA), total antioxidant status (TAOS), adiponectin, transforming growth factor (TGF-beta1), in occurence of NASH. Indones Biomed J. 2009; 1(1): 40-4, CrossRef.
Hou Y, Ding W, Wu P, Liu C, Ding L, Liu J, et al. Adipose-derived stem cells alleviate liver injury induced by type 1 diabetes mellitus by inhibiting mitochondrial stress and attenuating inflammation. Stem Cell Res Ther. 2022; 13(1): 132, CrossRef.
Gunes A, Schmitt C, Bilodeau L, Huet C, Belblidia A, Baldwin C, et al. IL-6 trans-signaling is increased in diabetes, impacted by glucolipotoxicity, and associated with liver stiffness and fibrosis in fatty liver disease. Diabetes. 2023; 72(12): 1820-34, CrossRef.
Ng CH, Chan KE, Chin YH, Zeng RW, Tsai PC, Lim WH, et al. The effect of diabetes and prediabetes on the prevalence, complications and mortality in nonalcoholic fatty liver disease. Clin Mol Hepatol. 2022; 28(3): 565-74, CrossRef.
Li B, Cheng Y, Yu S, Zang L, Yin Y, Liu J, et al. Human umbilical cord-derived mesenchymal stem cell therapy ameliorates nonalcoholic fatty liver disease in obese type 2 diabetic mice. Stem Cells Int. 2019; 2019: 8628027, CrossRef.
Ferguson D, Finck BN. Emerging therapeutic approaches for the treatment of NAFLD and type 2 diabetes mellitus. Nat Rev Endocrinol. 2021; 17(8): 484-95, CrossRef.
Wan Y, Garner J, Wu N, Phillip L, Han Y, McDaniel K, et al. Role of stem cells during diabetic liver injury. J Cell Mol Med. 2016; 20(2): 195-203, CrossRef.
Jiang R, Han Z, Zhuo G, Qu X, Li X, Wang X, et al. Transplantation of placenta-derived mesenchymal stem cells in type 2 diabetes: A pilot study. Front Med. 2011; 5(1): 94-100, CrossRef.
Skyler JS, Fonseca VA, Segal KR, Rosenstock J. Allogeneic mesenchymal precursor cells in type 2 diabetes: A randomized, placebo-controlled, dose-escalation safety and tolerability pilot study. Diabetes Care. 2015; 38(9): 1742-9, CrossRef.
Kong D, Zhuang X, Wang D, Qu H, Jiang Y, Li X, et al. Umbilical cord mesenchymal stem cell transfusion ameliorated hyperglycemia in patients with type 2 diabetes mellitus. Clin Lab. 2014; 60(12): 1969-76, CrossRef.
Liu P, Mao Y, Xie Y, Wei J, Yao J. Stem cells for treatment of liver fibrosis/cirrhosis: clinical progress and therapeutic potential. Stem Cell Res Ther. 2022; 13(1): 356, CrossRef.
Pawitan JA. Prospect of stem cell conditioned medium in regenerative medicine. Biomed Res Int. 2014: 2014: 965849, CrossRef.
Galipeau J. The mesenchymal stromal cells dilemma-does a negative phase III trial of random donor mesenchymal stromal cells in steroid-resistant graft-versus-host disease represent a death knell or a bump in the road? Cytotherapy. 2013; 15(1): 2-8, CrossRef.
Hendrawan S, Lheman J, Marcelina O, Tantoso L, Baer HU, Gunawan S. Regenerative therapeutic effects of conditioned medium from human umbilical cord‑derived mesenchymal stem cells as an adjuvant to insulin therapy in a rat model of type 2 diabetes. World Acad Sci J. 2025; 7(116): 1-13, CrossRef.
Hendrawan S, Lheman J, Dewi FNA, Nuraeni, Permanawati, Baer HU. Effect of mesenchymal stem cell conditioned medium on hepatocyte matrix implant to alleviate liver cirrhosis in rats. Regen Med. 2025; 20(10): 475-87, CrossRef.
Hendrawan S, Marcelina O, Tan ST, Baer HU. Immobilization of hUC-MSCs conditioned medium on 3D PLLA collagen-coated matrix enhances diabetic wound healing progression. Eng Regen. 2024; 5(3): 421-31, CrossRef.
Charan J, Kantharia ND. How to calculate sample size in animal studies? J Pharmacol Pharmacother. 2013; 4(4): 303-6, CrossRef.
Arifin WN, Zahiruddin WM. Sample size calculation in animal studies using resource equation approach. Malaysian J Med Sci. 2017; 24(5): 101-5, CrossRef.
Song D, Yin L, Wang C, Wen X. Adenovirus-mediated expression of SIK1 improves hepatic glucose and lipid metabolism in type 2 diabetes mellitus rats. PLoS One. 2019; 14(6): e0210930, CrossRef.
Song D, Yin L, Wang C, Wen X. Zhenqing recipe attenuates non-alcoholic fatty liver disease by regulating the SIK1/CRTC2 signaling in experimental diabetic rats. BMC Complement Med Ther. 2020; 20(1): 27, CrossRef.
Srinivasan K, Viswanad B, Asrat L, Kaul CL, Ramarao P. Combination of high-fat diet-fed and low-dose streptozotocin-treated rat: A model for type 2 diabetes and pharmacological screening. Pharmacol Res. 2005; 52(4): 313-20, CrossRef.
Gunawan S, Soetikno V, Purwaningsih E, Ferdinal F, Wuyung P, Ramadhani D. 6-Gingerol, a bioactive compound of Zingiber officinale, ameliorates high-fat high-fructose diet-induced non-alcoholic related fatty liver disease in rats. J Exp Pharmacol. 2024; 16: 455-66, CrossRef.
Liang W, Menke AL, Driessen A, Koek GH, Lindeman JH, Stoop R, et al. Establishment of a general NAFLD scoring system for rodent models and comparison to human liver pathology. Sookoian SC, editor. PLoS One. 2014; 9(12): e115922, CrossRef.
Standish RA. An appraisal of the histopathological assessment of liver fibrosis. Gut. 2006; 55(4): 569-78, CrossRef.
Oladipupo SO, Ezenabor EH, Ojo AB, Ogunlakin AD, Ojo OA. Interplay of the pathophysiological mechanisms of non-alcoholic fatty liver disease, diabetes mellitus, and inflammation: A growing threat to public health. Obes Med. 2025; 55: 100613, CrossRef.
Ahirwar B, Verma V, Godghate D, Parmar H. Prevalence and risk factors of non-alcoholic fatty liver disease (NAFLD) in type 2 diabetic patients. Eur J Cardiovasc Med. 2025; 15(6): 415-9, CrossRef.
Ginès P, Serra-Burriel M. Glucagon-like peptide-1 receptor agonists for treatment of steatotic liver disease in patients with type 2 diabetes mellitus: Growing evidence but not yet there. Gastroenterology. 2024; 167(4): 653-5, CrossRef.
ElSayed NA, Aleppo G, Aroda VR, Bannuru RR, Brown FM, Bruemmer D, et al. Comprehensive medical evaluation and assessment of comorbidities: Standards of care in diabetes-2023. Diabetes Care. 2022; 46 (Supplement 1): s49-s67.2, CrossRef.
Lu W, Qu J, Yan L, Tang X, Wang X, Ye A, et al. Efficacy and safety of mesenchymal stem cell therapy in liver cirrhosis: a systematic review and meta-analysis. Stem Cell Res Ther. 2023; 14(1): 301, CrossRef.
Sun Y, Shi H, Yin S, Ji C, Zhang X, Zhang B, et al. Human mesenchymal stem cell derived exosomes alleviate type 2 diabetes mellitus by reversing peripheral insulin resistance and relieving β-cell destruction. ACS Nano. 2018; 12(8): 7613-28, CrossRef.
Schaschkow A, Mura C, Dal S, Langlois A, Seyfritz E, Sookhareea C, et al. Impact of the Type of Continuous Insulin Administration on Metabolism in a Diabetic Rat Model. J Diabetes Res. 2016; 2016: 8310516, CrossRef.
Fennel ZJ, Kurian AS, Bourrant P, Skiles CM, Castro RJ, Yee EM, et al. Stem cell secretome treatment reduces adiposity and improves glucose handling during obesity and weight loss in mice. Obesity. 2026; 34(2): 382-93, CrossRef.
Mahmoudi SK, Abdolahi S, Saniee P, Zali MR, Hatami B, Baghaei K. Limosilactobacillus fermentum role in combination with human mesenchymal stem cell-derived secretome: A novel approach to alleviate inflammation in NASH pathogenesis. Int Immunopharmacol. 2025; 156: 114686, CrossRef.
Maimunah U, Kholili U, Putra RR, Brimantyo D, Wirantara H. Increased levels of TNF-α, IL-6, and IL-10 are associated with the degree of liver fibrosis in chronic hepatitis B patients with NUC therapy. Indones Biomed J. 2024; 16(1): 72-8, CrossRef.
Cuadra B, Silva V, Huang YL, Diaz Y, Rivas C, Molina C, et al. The immunoregulatory and regenerative potential of activated human stem cell secretome mitigates acute-on-chronic liver failure in a rat Model. Int J Mol Sci. 2024; 25(4): 2073, CrossRef.
Jing Y, Sun Q, Xiong X, Meng R, Tang S, Cao S, et al. Hepatocyte growth factor alleviates hepatic insulin resistance and lipid accumulation in high‐fat diet‐fed mice. J Diabetes Investig. 2019; 10(2): 251-60, CrossRef.
DOI: https://doi.org/10.18585/inabj.v18i4.4287
Copyright (c) 2026 The Prodia Education and Research Institute
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Indexed by:
The Prodia Education and Research Institute
