Skip to main content
SpringerLink
Log in

Therapeutic potential of Lactobacillus casei and Chlorella vulgaris in high-fat diet-induced non-alcoholic fatty liver disease (NAFLD)-associated kidney damages: a stereological study

  • Original Article
  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

Background

The non-alcoholic fatty liver disease (NAFLD) is prevalent in as many as 25% of adults who are afflicted with metabolic syndrome. Oxidative stress plays a significant role in the pathophysiology of hepatic and renal injury associated with NAFLD. Therefore, probiotics such as Lactobacillus casei (LBC) and the microalga Chlorella vulgaris (CV) may be beneficial in alleviating kidney injury related to NAFLD.

Materials and methods

This animal study utilized 30 C57BL/6 mice, which were evenly distributed into five groups: the control group, the NAFLD group, the NAFLD + CV group, the NAFLD + LBC group, and the NAFLD + CV + LBC group. A high-fat diet (HFD) was administered to induce NAFLD for six weeks. The treatments with CV and LBC were continued for an additional 35 days. Biochemical parameters, total antioxidant capacity (TAC), and the expression of kidney damage marker genes (KIM 1 and NGAL) in serum and kidney tissue were determined, respectively. A stereological analysis was conducted to observe the structural changes in kidney tissues.

Results

A liver histopathological examination confirmed the successful induction of NAFLD. Biochemical investigations revealed that the NAFLD group exhibited increased ALT and AST levels, significantly reduced in the therapy groups (p < 0.001). The gene expression levels of KIM-1 and NGAL were elevated in NAFLD but were significantly reduced by CV and LBC therapies (p < 0.001). Stereological examinations revealed reduced kidney size, volume, and tissue composition in the NAFLD group, with significant improvements observed in the treated groups (p < 0.001).

Conclusion

This study highlights the potential therapeutic efficacy of C. vulgaris and L. casei in mitigating kidney damage caused by NAFLD. These findings provide valuable insights for developing novel treatment approaches for managing NAFLD and its associated complications.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Data availability

This article contains all data created and examined throughout this investigation. The corresponding author will provide datasets used or analyzed during the current work upon reasonable request.

Abbreviations

ALT:

Alanine Aminotransferase

AST:

Aspartate Aminotransferase

BUN:

Blood Urea Nitrogen

Chol:

Cholesterol

CKD:

Chronic Kidney Disease

CV:

Chlorella vulgaris

CVD:

Cardio vascular Diseases

DCT:

Distal convoluted tubule

FOXO1:

Forkhead box protein O1

FRAP:

Ferric reducing Antioxidant Power

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

H&E:

Hematoxylin and Eosin

HFD:

High-fat diet

KIM-1:

Kidney Injury Molecule-1

LBC:

Lactobacillus casei

MAFLD:

Metabolic-Associated Fatty Liver disease

NAFLD:

Non-alcoholic fatty liver disease

NASH:

Non-alcoholic steatohepatitis

NGAL:

Neutrophil gelatinase-associated lipocalin

OS:

Oxidative stress

p-AMPK:

Phosphorylated AMP-activated protein kinase

PCT:

Proximal convoluted tubule

ROS:

Reactive Oxygen Species

RT-PCR:

Real-Time Polymerase Chain Reaction

T2D:

Type 2 diabetes

TAC:

Total Antioxidant Capacity

TG:

Triglyceride

TPTZ:

2,4,6 tripyridyl-s-triazine

References

  1. Choi J, Joe H, Oh J-E, Cho Y-J, Shin H-S, Heo NH (2023) The correlation between NAFLD and serum uric acid to serum creatinine ratio. PLoS ONE 18:e0288666

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Pouwels S, Sakran N, Graham Y et al (2022) Non-alcoholic fatty liver disease (NAFLD): a review of pathophysiology, clinical management and effects of weight loss. BMC Endocr Disorders 22:1–9

    Article  Google Scholar 

  3. Pai RK (2019) NAFLD histology: a critical review and comparison of scoring systems. Curr Hepatol Rep 18:473–481

    Article  Google Scholar 

  4. Zheng X, Cao C, He Y, Wang X, Wu J, Hu H (2021) Association between nonalcoholic fatty liver disease and incident diabetes mellitus among Japanese: a retrospective cohort study using propensity score matching. Lipids Health Dis 20:59

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Zhang M, Lin S, Wang M-f et al (2020) Association between NAFLD and risk of prevalent chronic kidney disease: why there is a difference between east and west? BMC Gastroenterol 20:1–7

    Article  Google Scholar 

  6. Mantovani A, Lombardi R, Cattazzo F, Zusi C, Cappelli D, Dalbeni A (2022) MAFLD and CKD: an updated narrative review. Int J Mol Sci 23:7007–7007

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Lonardo A, Mantovani A, Targher G, Baffy G (2022) Nonalcoholic fatty liver disease and chronic kidney disease: epidemiology, pathogenesis, and clinical and research implications. Int J Mol Sci 23:13320–13320

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Hydes T, Buchanan R, Kennedy OJ, Fraser S, Parkes J, Roderick P (2020) Systematic review of the impact of non-alcoholic fatty liver disease on mortality and adverse clinical outcomes for individuals with chronic kidney disease. BMJ Open 10:e040970–e040970

    Article  PubMed  PubMed Central  Google Scholar 

  9. Gonzalez A, Huerta-Salgado C, Orozco-Aguilar J et al (2020) Role of oxidative stress in hepatic and extrahepatic dysfunctions during nonalcoholic fatty liver Disease (NAFLD). Oxidative Med Cell Longev 2020:1617805–1617805

    Article  Google Scholar 

  10. Delli Bovi AP, Marciano F, Mandato C, Siano MA, Savoia M, Vajro P (2021) Oxidative stress in non-alcoholic fatty liver disease. An updated mini review. Front Med 8:165–165

    Article  Google Scholar 

  11. Nielsen S, Schjoedt K, Astrup A et al (2009) Kidney injury molecule 1 (KIM1) and neutrophil gelatinase-associated lipocalin (NGAL) as markers of tubular kidney injury in type 1 diabetic patients

  12. Abourady AA, Amin SM, Kasem SM, Abdelnaby HH (2020) Measurement of urinary kidney injury molecule-1 level as an early biomarker of renal impairment in overweight/obese children and adolescents with nonalcoholic fatty liver disease. J Am Sci 16

  13. Chen L, Lv X, Kan M, Wang R, Wang H, Zang H (2022) Critical overview of hepatic factors that link non-alcoholic fatty liver disease and acute kidney Injury: physiology and therapeutic implications. Int J Mol Sci 23:12464

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Francoz C, Nadim MK, Durand F (2016) Kidney biomarkers in cirrhosis. J Hepatol 65:809–824

    Article  CAS  PubMed  Google Scholar 

  15. David D, Eapen CE (2021) What are the current pharmacological therapies for nonalcoholic fatty liver disease? J Clin Experimental Hepatol 11:232–238

    Article  CAS  Google Scholar 

  16. Shao Y, Chen S, Han L, Liu J (2023) Pharmacotherapies of NAFLD: updated opportunities based on metabolic intervention. Nutr Metabolism 20:30

    Article  CAS  Google Scholar 

  17. Abdella A, Elbadawy MF, Irmak S, Alamri ES (2022) Hypolipidemic, antioxidant and Immunomodulatory effects of Lactobacillus casei ATCC 7469-Fermented wheat Bran and Spirulina maxima in rats Fed a High-Fat Diet. Fermentation 8:610–610

    Article  CAS  Google Scholar 

  18. Lee NY, Shin MJ, Youn GS et al (2021) Lactobacillus attenuates progression of nonalcoholic fatty liver disease by lowering cholesterol and steatosis. Clin Mol Hepatol 27:110–110

    Article  PubMed  Google Scholar 

  19. Machu L, Misurcova L, Vavra Ambrozova J et al (2015) Phenolic content and antioxidant capacity in algal food products. Molecules 20:1118–1133

    Article  PubMed  PubMed Central  Google Scholar 

  20. Wan M, Li Q, Lei Q, Zhou D, Wang S (2022) Polyphenols and polysaccharides from morus alba l. fruit attenuate high-fat diet-induced metabolic syndrome modifying the gut microbiota and metabolite profile. Foods 11:1818–1818

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Panahi Y, Ghamarchehreh ME, Beiraghdar F, Zare R, Jalalian HR, Sahebkar A (2012) Investigation of the effects of Chlorella vulgaris supplementation in patients with non-alcoholic fatty liver disease: a randomized clinical trial. Hepatogastroenterology 59:2099–2103

    PubMed  Google Scholar 

  22. Mohseni R, Alavian SM, Sadeghabadi ZA, Heiat M (2021) Therapeutic effects of Chlorella vulgaris on carbon tetrachloride induced liver fibrosis by targeting Hippo signaling pathway and AMPK/FOXO1 axis. Mol Biol Rep 48:117–126

    Article  CAS  PubMed  Google Scholar 

  23. An H-J, Choi H-M, Park H-S et al (2006) Oral administration of hot water extracts of Chlorella vulgaris increases physical stamina in mice. Annals Nutr Metabolism 50:380–386

    Article  CAS  Google Scholar 

  24. Zhang Z, Zhou H, Guan M et al (2021) Lactobacillus casei YRL577 combined with plant extracts reduce markers of non-alcoholic fatty liver disease in mice. Br J Nutr 125:1081–1091

    Article  CAS  PubMed  Google Scholar 

  25. Karp NA, Meehan TF, Morgan H et al (2015) Applying the ARRIVE guidelines to an in vivo database. PLoS Biol 13, e1002151

  26. Cho J, Lee I, Kim D et al (2014) Effect of aerobic exercise training on non-alcoholic fatty liver disease induced by a high fat diet in C57BL/6 mice. J Exerc Nutr Biochem 18:339

    Article  Google Scholar 

  27. Zhang Z, Zhou H, Zhou X et al (2021) Lactobacillus casei YRL577 ameliorates markers of non-alcoholic fatty liver and alters expression of genes within the intestinal bile acid pathway. Br J Nutr 125:521–529

    Article  CAS  PubMed  Google Scholar 

  28. Lim CSH, Lim SL (2013) Ferric reducing capacity versus ferric reducing antioxidant power for measuring total antioxidant capacity. Lab Med 44:51–55

    Article  Google Scholar 

  29. Scherle W (1970) A simple method for volumetry of organs in quantitative stereology. Mikroskopie 26:57–60

    CAS  PubMed  Google Scholar 

  30. Maleki MH, Nadimi E, Vakili O et al (2023) Bilirubin improves renal function by reversing the endoplasmic reticulum stress and inflammation in the kidneys of type 2 diabetic rats fed high-fat diet. Chemico-Biol Interact 378:110490

    Article  CAS  Google Scholar 

  31. Roderburg C, Krieg S, Krieg A et al (2023) Non-alcoholic fatty liver disease (NAFLD) is associated with an increased incidence of chronic kidney disease (CKD). Eur J Med Res 28:153–153

    Article  PubMed  PubMed Central  Google Scholar 

  32. Ebrahimi-Mameghani M, Aliashrafi S, Javadzadeh Y, AsghariJafarabadi M (2014) The effect of Chlorella vulgaris supplementation on liver enzymes, serum glucose and lipid profile in patients with non-alcoholic fatty liver disease. Health Promotion Perspect 4:107

    Google Scholar 

  33. Khani S, Hosseini M, Taheri H, Nourani MR M and, Imani Fooladi A (2012) A Probiotics as an alternative strategy for prevention and treatment of human diseases: a review. Inflammation & Allergy-Drug Targets (Formerly Current Drug Targets-Inflammation & Allergy)(Discontinued) 11, 79–89

  34. Fagundes RAB, Soder TF, Grokoski KC, Benetti F, Mendes RH (2018) Probiotics in the treatment of chronic kidney disease: a systematic review. Brazilian J Nephrol 40:278–286

    Article  Google Scholar 

  35. Elmeleh MI, Attia T, Elgendy H (2023) Protective effect of Chlorella vulgaris and Spirulina platensis against ThioacetamideInduced Hepatorenal toxicity in male rats. J Curr Veterinary Res 5:79–92

    Article  Google Scholar 

  36. Regueiras A, Huguet Á, Conde T et al (2021) Potential anti-obesity, anti-steatosis, and anti-inflammatory properties of extracts from the microalgae Chlorella vulgaris and Chlorococcum amblystomatis under different growth conditions. Mar Drugs 20:9

    Article  PubMed  PubMed Central  Google Scholar 

  37. Andy SY, Keeffe EB (2003) Elevated AST or ALT to nonalcoholic fatty liver disease. accurate predictor of disease prevalence?

  38. Imamura Y, Mawatari S, Oda K et al (2021) Changes in body composition and low blood urea nitrogen level related to an increase in the prevalence of fatty liver over 20 years: a cross-sectional study. Hepatol Res 51:570–579

    Article  CAS  PubMed  Google Scholar 

  39. Gonzalez A, Huerta-Salgado C, Orozco-Aguilar J et al (2020) Role of oxidative stress in hepatic and extrahepatic dysfunctions during nonalcoholic fatty liver disease (NAFLD). Oxidative Medicine and Cellular Longevity 2020

  40. Aron-Wisnewsky J, Warmbrunn MV, Nieuwdorp M, Clement K (2020) Nonalcoholic fatty liver disease: modulating gut microbiota to improve severity? Gastroenterology 158:1881–1898

    Article  CAS  PubMed  Google Scholar 

  41. Karlsson C, Lindell K, Ottosson M, Sjöström L, Carlsson Br, Carlsson LMS (1998) Human adipose tissue expresses angiotensinogen and enzymes required for its Conversion to Angiotensin II1. J Clin Endocrinol Metabolism 83:3925–3929

    CAS  Google Scholar 

  42. Yang M, Geng C-A, Liu X, Guan M (2021) Lipid disorders in NAFLD and chronic kidney disease. Biomedicines 9:1405–1405

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Yang M, Geng C-A, Liu X, Guan M (2021) Lipid disorders in NAFLD and chronic kidney disease. Biomedicines 9:1405

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Zhou D, Pan Q, Shen F et al (2017) Total fecal microbiota transplantation alleviates high-fat diet-induced steatohepatitis in mice via beneficial regulation of gut microbiota. Sci Rep 7:1529

    Article  PubMed  PubMed Central  Google Scholar 

  45. Wagnerberger S, Spruss A, Kanuri G et al (2013) Lactobacillus casei Shirota protects from fructose-induced liver steatosis: a mouse model. J Nutr Biochem 24:531–538

    Article  CAS  PubMed  Google Scholar 

  46. Song H, Han W, Yan F, Xu D, Chu Q, Zheng X (2016) Dietary Phaseolus vulgaris extract alleviated diet-induced obesity, insulin resistance and hepatic steatosis and alters gut microbiota composition in mice. J Funct Foods 20:236–244

    Article  CAS  Google Scholar 

  47. Wan X, Li T, Liu D et al (2018) Effect of marine microalga Chlorella pyrenoidosa ethanol extract on lipid metabolism and gut microbiota composition in high-fat diet-fed rats. Mar Drugs 16:498

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Lin L, Tan W, Pan X, Tian E, Wu Z, Yang J (2022) Metabolic syndrome-related kidney injury: a review and update. Front Endocrinol 13:904001–904001

    Article  Google Scholar 

  49. Wanchai K, Yasom S, Tunapong W et al (2018) Probiotic Lactobacillus paracasei HII01 protects rats against obese-insulin resistance-induced kidney injury and impaired renal organic anion transporter 3 function. Clin Sci 132:1545–1563

    Article  CAS  Google Scholar 

  50. Zhang W, Miikeda A, Zuckerman J et al (2021) Inhibition of microbiota-dependent TMAO production attenuates chronic kidney disease in mice. Sci Rep 11:518–518

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  51. Zhu H, Cao C, Wu Z et al (2021) The probiotic L. Casei Zhang slows the progression of acute and chronic kidney disease. Cell Metabol 33:1926–1942

    Article  CAS  Google Scholar 

  52. Lee HS, Park HJ, Kim MK (2008) Effect of Chlorella vulgaris on lipid metabolism in Wistar rats fed high fat diet. Nutr Res Pract 2:204–210

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  53. Mounir M, Ibijbijen A, Farih K, Rabetafika HN, Razafindralambo HL (2022) Synbiotics and their antioxidant properties, mechanisms, and benefits on human and animal health: a narrative review. Biomolecules 12:1443

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  54. Li Q, Zheng T, Ding H et al (2023) Exploring the benefits of Probiotics in Gut inflammation and Diarrhea—from an antioxidant perspective. Antioxidants 12:1342

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Kleniewska P, Hoffmann A, Pniewska E, Pawliczak R (2016) The influence of probiotic Lactobacillus casei in combination with prebiotic inulin on the antioxidant capacity of human plasma. Oxidative medicine and cellular longevity 2016

  56. Keshavarz Azizi Raftar S, Ashrafian F, Yadegar A et al (2021) The protective effects of live and pasteurized Akkermansia muciniphila and its extracellular vesicles against HFD/CCl4-induced liver injury. Microbiol Spectr 9:e00484–e00421

    Article  PubMed  PubMed Central  Google Scholar 

  57. Altunkaynak ME, Özbek E, Altunkaynak BZ, Can İ, Unal D, Unal B (2008) The effects of high-fat diet on the renal structure and morphometric parametric of kidneys in rats. J Anat 212:845–852

    Article  PubMed  PubMed Central  Google Scholar 

  58. Armitage JA, Lakasing L, Taylor PD et al (2005) Developmental programming of aortic and renal structure in offspring of rats fed fat-rich diets in pregnancy. J Physiol 565:171–184

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  59. Schulz C-A, Engström G, Nilsson J et al (2020) Plasma kidney injury molecule-1 (p-KIM-1) levels and deterioration of kidney function over 16 years. Nephrol Dialysis Transplantation 35:265–273

    Article  CAS  Google Scholar 

  60. Allegretti AS, Parada XV, Endres P et al (2021) Urinary NGAL as a diagnostic and prognostic marker for acute kidney injury in cirrhosis: a prospective study. Clinical and translational gastroenterology 12

Download references

Acknowledgements

The present article was extracted from the thesis written by Haniyeh Keyghobadi has been submitted in Department of Biology, Zarghan Branch, Islamic Azad University, Zarghan, Iran.

Funding

Public, private nor nonprofit funding organizations did not provide any specific grants for this research.

Author information

Authors and Affiliations

  1. Department of Biology, Zarghan Branch, Islamic Azad University, Zarghan, Iran

    Haniyeh Keyghobadi & Hadis bozorgpoursavadjani

  2. Endocrinology and Metabolism Research Center, Shiraz University of Medical Sciences, Shiraz, Iran

    Farhad Koohpeyma, Marzieh Nemati & Sanaz Dastghaib

  3. Student Research Committee, Shiraz University of Medical Sciences, Shiraz, Iran

    Farhad Koohpeyma & Farshad Dehghani

  4. Nutrition and Integrative Physiology, Florida State University, Tallahassee, FL, USA

    Nazanin Mohammadipoor

  5. Molecular Dermatology Research Center, Shiraz University of Medical Sciences, Shiraz, Iran

    Iman Jamhiri

  6. Deparment of Agriculture Management, Shiraz Branch, Islamic Azad University, Shiraz, Iran

    Gholamhossein Keighobadi

  7. Autophagy Research Center, Shiraz University of Medical Sciences, Shiraz, Iran

    Sanaz Dastghaib

Authors
  1. Haniyeh Keyghobadi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hadis bozorgpoursavadjani
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Farhad Koohpeyma
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Nazanin Mohammadipoor
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Marzieh Nemati
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Farshad Dehghani
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Iman Jamhiri
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Gholamhossein Keighobadi
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Sanaz Dastghaib
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

All experiments, statistical analysis, and figure preparation were conducted by H.K (Haniyeh Keyghobadi), H.B (Hadis bozorgpoursavadjani), N.M (Nazanin Mohammadipoor) and F.K (Farhad Koohpeyma). The initial draft of manuscript was written by M.N (Marzieh Nemati), F.D (Farshad Dehghani), and GH.K (Gholamhossein Keighobadi). All tests were set up and second draft of manuscript was written by F.k, and SD (Sanaz Dastghaib). A final manuscript proof was completed by SD who also contributed more funds to the project.

Corresponding author

Correspondence to Sanaz Dastghaib.

Ethics declarations

Ethics approval and consent to participate

The Research Ethics Committee of the Islamic Azad University-Arsanjan Branch (Fars Province) has ethically confirmed the study (Ethics Code: IR.IAU.A.REC.1402.065).

Consent for publication

Not applicable.

Conflict of interest

Authors affirm that they don’t have competitive interests.

Additional information

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Material 1

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Keyghobadi, H., bozorgpoursavadjani, H., Koohpeyma, F. et al. Therapeutic potential of Lactobacillus casei and Chlorella vulgaris in high-fat diet-induced non-alcoholic fatty liver disease (NAFLD)-associated kidney damages: a stereological study. Mol Biol Rep 51, 613 (2024). https://doi.org/10.1007/s11033-024-09542-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11033-024-09542-1

Keywords

Search

Navigation