Annals of Microbiology

, Volume 69, Issue 5, pp 483–494 | Cite as

Milk fermented with probiotic strains Lactobacillus rhamnosus MTCC: 5957 and Lactobacillus rhamnosus MTCC: 5897 ameliorates the diet-induced hypercholesterolemia in rats

  • Radha YadavEmail author
  • Rishika Vij
  • Suman Kapila
  • Suhail H. Khan
  • Naveen Kumar
  • Sunita Meena
  • Rajeev Kapila
Original Article


The current study was intended to investigate the cholesterol-lowering potential of the two Lactobacillus rhamnosus probiotic strains, LR 5957 and LR 5897, isolated from ‘dahi’. Cholesterol-lowering ability of both strains was determined in in vitro conditions. For in vivo investigations, the Wistar rats were randomly assigned into five groups and treated with different diets: standard diet (SD), high-cholesterol diet (HCD), HCD with Milk, HCD with LR 5957–fermented milk, and HCD with LR 5897–fermented milk. After 3 months of feeding, different parameters of hypercholesterolemia were measured in blood, feces, liver, and kidney. Both the strains, LR 5957 and LR 5897, showed the ability to grow in the presence of cholesterol and eliminate the cholesterol under in vitro conditions. In vivo results indicate that consumption of probiotic-fermented milk has significantly reduced the HCD-induced body weight, hyperlipidemia, and hepatic lipids (total cholesterol and triacylglycerol). Further, increased cholesterol excretion in feces was also observed in probiotic-fed groups. The studied fermented milk also helped to maintain healthy liver and kidney by increasing the antioxidant activities and decreasing the lipid peroxidation. Consumption of probiotic-fermented milk also found to decrease the mRNA expression of the inflammatory markers TNF-α and IL-6 in the liver. Overall, our results indicate that the L. rhamnosus strains, LR 5957 and LR 5897, are two potential probiotic strains that can ameliorate the diet-induced hypercholesterolemia.


Probiotics Hypercholesterolemia Probiotic-fermented milk Lactobacillus rhamnosus 



The authors are grateful to ICAR-National Dairy Research Institute, Karnal, for providing funding and laboratory facilities to carry out this work.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical statement

The study was approved by the Institute Animal Ethical Committee (IAEC) (IAEC No. 101/16 dated 21.04.2016) of Indian Council of Agriculture Research-National Dairy Research Institute.

Informed consent was obtained from all individual participants included in the study.


  1. Aebi H (1984) [13] Catalase in vitro. Methods Enzymol 105:121–126CrossRefGoogle Scholar
  2. Al-Sheraji SH, Ismail A, Manap MY, Mustafa S, Yusof RM, Hassan FA (2012) Hypocholesterolaemic effect of yoghurt containing Bifidobacterium pseudocatenulatum G4 or Bifidobacterium longum BB536. Food Chem 135(2):356–361CrossRefGoogle Scholar
  3. AOAC (1990) Official methods of analysis, 16th edn. Association of Official Agricultural Chemists, Washington, DCGoogle Scholar
  4. Boers M, Nurmohamed MT, Doelman CJ, Lard LR, Verhoeven AC, Voskuyl AE, Huizinga TW, Van de Stadt RJ, Dijkmans BA, van der Linden S (2003) Influence of glucocorticoids and disease activity on total and high density lipoprotein cholesterol in patients with rheumatoid arthritis. Ann Rheum Dis 62:842–845CrossRefGoogle Scholar
  5. Brown MS, Goldstein JL (1986) A receptor-mediated pathway for cholesterol homeostasis. Science 232(4746):34–47CrossRefGoogle Scholar
  6. Choi EA, Chang HC (2015) Cholesterol-lowering effects of a putative probiotic strain Lactobacillus plantarum EM isolated from kimchi. LWT-Food Sci Technol 62(1):210–217CrossRefGoogle Scholar
  7. Coppack SW (2001) Pro-inflammatory cytokines and adipose tissue. Proc Nutr 60(3):349–356CrossRefGoogle Scholar
  8. Dai C, Zhao DH, Jiang M (2012) VSL# 3 probiotics regulate the intestinal epithelial barrier in vivo and in vitro via the p38 and ERK signaling pathways. Int J Mol Med 29(2):202–208Google Scholar
  9. Ewaschuk J, Endersby R, Thiel D, Diaz H, Backer J, Ma M, Churchill T, Madsen K (2007) Probiotic bacteria prevent hepatic damage and maintain colonic barrier function in a mouse model of sepsis. Hepatology 46(3):841–850CrossRefGoogle Scholar
  10. Folch J, Lees M, Sloane-Stanley GH (1957) A simple method for the isolation and purification of total lipids from animal tissues. J Biol Chem 226(1):497–509Google Scholar
  11. Friedewald WT, Levy RI, Fredrickson DS (1972) Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge. Clin Chem 18(6):499–502Google Scholar
  12. Gielen S, Landmesser U (2014) The year in cardiology 2013: cardiovascular disease prevention. Eur Heart J 35(5):307–312CrossRefGoogle Scholar
  13. Goldstein JL, Brown MS (1990) Regulation of the mevalonate pathway. Nature 343(6257):425CrossRefGoogle Scholar
  14. Huang Y, Wang X, Wang J, Wu F, Sui Y, Yang L, Wang Z (2013) Lactobacillus plantarum strains as potential probiotic cultures with cholesterol-lowering activity. J Dairy Sci 96(5):2746–2753CrossRefGoogle Scholar
  15. Hung SC, Tseng WT, Pan TM (2016) Lactobacillus paracasei subsp. paracasei NTU 101 ameliorates impaired glucose tolerance induced by a high-fat, high-fructose diet in Sprague-Dawley rats. J Funct Foods 24:472–481CrossRefGoogle Scholar
  16. Kalyan S, Meena S, Kapila S, Sowmya K, Kumar R (2018) Evaluation of goat milk fat and goat milk casein fraction for anti-hypercholesterolaemic and antioxidative properties in hypercholesterolaemic rats. Int Dairy J 84:23–27CrossRefGoogle Scholar
  17. Kaushal D, Kansal VK (2012) Probiotic dahi containing Lactobacillus acidophilus and Bifidobacteriumbifidum alleviates age-inflicted oxidative stress and improves expression of biomarkers of ageing in mice. Mol Biol Rep 39(2):1791–1799CrossRefGoogle Scholar
  18. Kemgang TS, Kapila S, Shanmugam VP, Reddi S, Kapila R (2016) Fermented milk with probiotic Lactobacillus rhamnosus S1K3 (MTCC5957) protects mice from salmonella by enhancing immune and nonimmune protection mechanisms at intestinal mucosal level. J Nutr Biochem 30:62–73CrossRefGoogle Scholar
  19. Kimoto H, Ohmomo S, Okamoto T (2002) Cholesterol removal from media by lactococci. J Dairy Sci 85(12):3182–3188CrossRefGoogle Scholar
  20. Kumar M, Shruti R, Ravinder N, Hemalatha RA, Sudarshan V, Ramagoni R (2013) Probiotic Lactobacillus rhamnosus GG and aloe vera gel improve lipid profiles in hypercholesterolemic rats. Nutrition 29(3):574–579CrossRefGoogle Scholar
  21. Kumar N, Tomar SK, Thakur K, Singh AK (2017) The ameliorative effects of probiotic Lactobacillus fermentum strain RS-2 on alloxan induced diabetic rats. J Funct Foods 28:275–284CrossRefGoogle Scholar
  22. Levine GN, Keaney JF, Vita JA (1995) Cholesterol reduction in cardiovascular disease—clinical benefits and possible mechanisms. N Engl J Med 332(8):512–521CrossRefGoogle Scholar
  23. Liong MT, Shah NP (2005) Acid and bile tolerance and cholesterol removal ability of lactobacilli strains. J Dairy Sci 88(1):55–66CrossRefGoogle Scholar
  24. Liu CS, Lin CC, Li TC (1999) The relation of white blood cell counts and athero- genic index ratio of LDL-cholesterol to HDL-cholesterol in Taiwan school children. Acta Paediatr Taiwanica 40:319–324Google Scholar
  25. Liu Y, Zhao F, Liu J, Wang H, Han X, Zhang Y, Yang Z (2017) Selection of cholesterol-lowering lactic acid bacteria and its effects on rats fed with high-cholesterol diet. Curr Microbiol 74(5):623–631CrossRefGoogle Scholar
  26. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193(1):265–275Google Scholar
  27. Marklund S, Marklund G (1974) Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. FEBS J 47(3):469–474Google Scholar
  28. Menard S, Candalh C, Bambou JC, Terpend K, Cerf-Bensussan N, Heyman M (2004) Lactic acid bacteria secrete metabolites retaining anti-inflammatory properties after intestinal transport. Gut 53(6):821–828CrossRefGoogle Scholar
  29. Michael DR, Davies TS, Moss JW, Calvente DL, Ramji DP, Marchesi JR, Pechlivanis A, Plummer SF, Hughes TR (2017) The anti-cholesterolaemic effect of a consortium of probiotics: an acute study in C57BL/6J mice. Sci Rep 7(1):2883CrossRefGoogle Scholar
  30. Osterberg KL, Boutagy NE, McMillan RP, Stevens JR, Frisard MI, Kavanaugh JW, Hulver MW (2015) Probiotic supplementation attenuates increases in body mass and fat mass during high-fat diet in healthy young adults. Obesity 23(12):2364–2370CrossRefGoogle Scholar
  31. Paglia DE, Valentine WN (1967) Studies on the quantitative and qualitative characterization of erythrocyte glutathione peroxidase. J Lab Clin Med 70(1):158–169Google Scholar
  32. Salaj R, Štofilová J, Šoltesová A, Hertelyová Z, Hijová E, Bertková I, Strojný L, Kružliak P, Bomba A (2013) The effects of two Lactobacillus plantarum strains on rat lipid metabolism receiving a high fat diet. Sci World J 2013:135142Google Scholar
  33. Saliganti V, Kapila R, Sharma R, Kapila S (2015) Feeding probiotic Lactobacillus rhamnosus (MTCC 5897) fermented milk to suckling mothers alleviates ovalbumin-induced allergic sensitisation in mice offspring. Br J Nutr 114(8):1168–1179CrossRefGoogle Scholar
  34. Sanchez M, Darimont C, Drapeau V, Emady-Azar S, Lepage M, Rezzonico E, Ngom-Bru C, Berger B, Philippe L, Ammon-Zuffrey C, Leone P (2014) Effect of Lactobacillus rhamnosus CGMCC1. 3724 supplementation on weight loss and maintenance in obese men and women. Br J Nutr 111(8):1507–1519CrossRefGoogle Scholar
  35. Sharma R, Kapila R, Dass G, Kapila S (2014) Improvement in Th1/Th2 immune homeostasis, antioxidative status and resistance to pathogenic E coli on consumption of probiotic Lactobacillus rhamnosus fermented milk in aging mice. Age 36(4):9686CrossRefGoogle Scholar
  36. Xiao JZ, Kondo S, Takahashi N, Miyaji K, Oshida K, Hiramatsu A, Iwatsuki K, Kokubo S, Hosono A (2003) Effects of milk products fermented by Bifidobacteriumlongum on blood lipids in rats and healthy adult male volunteers. J Dairy Sci 86(7):2452–2461CrossRefGoogle Scholar
  37. Yan F, Cao H, Cover TL, Whitehead R, Washington MK, Polk DB (2007) Soluble proteins produced by probiotic bacteria regulate intestinal epithelial cell survival and growth. Gastroenterology 132(2):562–575CrossRefGoogle Scholar
  38. Yonejima Y, Ushida K, Mori Y (2013) Lactobacillus gasseri NT decreased visceral fat through enhancement of lipid excretion in feces of KK-A y mice. Biosci Biotechnol Biochem 77(11):2312–2315CrossRefGoogle Scholar
  39. Yoo SR, Kim YJ, Park DY, Jung UJ, Jeon SM, Ahn YT, Choi MS (2013) Probiotics L. plantarum and L. curvatus in combination alter hepatic lipid metabolism and suppress diet-induced obesity. Obesity 21(12):2571–2578CrossRefGoogle Scholar
  40. Zhang XL, Wu YF, Wang YS, Wang XZ, Piao CH, Liu JM, Wang YH (2017) The protective effects of probiotic-fermented soymilk on high-fat diet-induced hyperlipidemia and liver injury. J Funct Foods 30:220–227CrossRefGoogle Scholar

Copyright information

© Università degli studi di Milano 2019

Authors and Affiliations

  • Radha Yadav
    • 1
    Email author
  • Rishika Vij
    • 1
  • Suman Kapila
    • 1
  • Suhail H. Khan
    • 1
  • Naveen Kumar
    • 1
  • Sunita Meena
    • 1
  • Rajeev Kapila
    • 1
  1. 1.Animal Biochemistry DivisionNational Dairy Research InstituteKarnalIndia

Personalised recommendations