Beneficial Effect of Lactiplantibacillus pentosus on Oxidative Stress and Histopathology in High-Fat-Induced Mice

 

Susmiati Susmiati1*, Sri Melia2, Ilfa Khairina1, Alimuddin Tofrizal3

1Department of Basic and Fundamental Nursing, Faculty of Nursing, Universitas Andalas,

Jl. Kampus Limau Manis Padang 25163, West Sumatra, Indonesia.

2Department of Animal Product Technology, Faculty of Animal Science, Universitas Andalas,

Jl. Kampus Limau Manis Padang 25163, Indonesia.

3Department of Anatomical Pathology, Faculty of Medicine, Universitas Andalas,

Jl. Kampus Limau Manis Padang 25163, Indonesia.

*Corresponding Author E-mail: susmiati@nrs.unand.ac.id

 

ABSTRACT:

Probiotics have been widely used to treat lipid metabolism disorders associated with obesity by modifying the microbiota balance. Probiotics play a role in health by maintaining intestinal homeostasis, alienating pathogens, increasing nutrient bioavailability, and stimulating and modulating the immune system. Various local probiotics have also been developed as a source of probiotics to assist with health issues. One is the probiotic Lactoplantibacilus pentosus, which comes from dadih (traditional fermented buffalo milk from West Sumatera ). This research aimed to assess the impact of Lactoplantibacilus pentosus on oxidative stress, adipose tissue, and liver histopathology in mice fed a high-fat diet (HFD). Methods: The experimental setup involved dividing mice into four groups, each consisting of seven animals. These groups were randomly assigned to one of four dietary regimens: ND (Normal Diet), HFD (High Fat Diet), HFDL (High Fat Diet supplemented with L. pentosus at 109 CFU/ml), and HDLFL (High Fat Diet supplemented with fermented milk containing L. pentosus at 109 CFU/ml). In the HFDL and HDLFL groups, L. pentosus was delivered orally once a day for six weeks at a 1 x 109 CFU/mL dosage. Result: Following a 6-week high-fat diet, the HFD group had a 24.30% higher body weight than ND group. The HDLFL group exhibited a lower body weight (23.78±0.84g) compared to the HFD group (26.59 ± 1.17g), with a statistically significant difference (p<0.05). Animals given probiotics had higher antioxidant SOD levels and lower MDA levels than the HFD group. Adipocyte hypertrophy, observed in animals on a high-fat diet, was mitigated by the administration of fermented milk and Lactobacillus pentosus. The HFD group exhibited higher levels of steatosis, inflammation, fibrosis, and necrosis in the liver compared to the normal diet group. Steatosis, inflammation, necrosis, and fibrotic scores decreased in the HFD group given Lactiplantibacillus pentosus fermented milk. Conclusion: Lactobacillus pentosus derived from dadih has been shown to reduce obesity, hepatic steatosis, and oxidative stress in people who regularly consume high-fat diets.

 

KEYWORDS: Hepatic steatosis, Lactobacillus pentosus, Obesity, Oxidative stress.

 

 


INTRODUCTION: 

Obesity and related metabolic diseases are caused by a variety of variables, including intrauterine and postnatal development, genetic susceptibility, poor food, socioeconomic level, physical activity, and hormone imbalances1.

 

A high-fat diet and sedentary lifestyle most commonly induce obesity. High-fat diets have been linked to an elevated risk of disorders such as hypertension, dislipidemia, and atherosclerosis, according to epidemiological studies2. Excessive consumption of fats can increase intestinal permeability, leading to varying levels of inflammation. In 2021, more than 1 billion people were classified as obese. WHO research predicts that this number will rise by 167 million by 2025. Obesity is the leading cause of T2D and prediabetes in children and adolescents, as well as elevated glycometabolic indicator levels3. Identifying these relationships may provide better guidance for managing obesity and related disorders. The prevention and management of metabolic illnesses associated with obesity must be thoroughly investigated. Effective obesity prevention is crucial for reducing the prevalence of cardiovascular and metabolic diseases4.

 

The primary methods for treating obesity include dietary control, exercise, surgical treatment, and pharmacological management. Among these, surgical and pharmacological treatments have been linked to severe adverse effects5. Numerous reviews have explored the development of anti-obesity medications6. Dietary modifications, such as calorie restriction and nutraceutical supplementation, are crucial in treating obesity. Caloric restriction (CR) is a dietary strategy that reduces caloric consumption and is an efficient way of weight loss. It influences adipose tissue remodelling and alters the endocrinological function of adiposity tissue and musculoskeletal7. Excess fat accumulation induces fat tissue proliferation, altering the secretory system and metabolites released, thereby affecting the surrounding microenvironment6.

 

Numerous studies have indicated that intestinal bacteria play a crucial role in adiposity and metabolic disorders, and it appears that obesity is directly related to changing gut microbiota8. Evidence indicates that the gut microbiota may be a therapeutic target for metabolic diseases9,10. Probiotics have recently emerged as a viable treatment for lipid metabolism disorders associated with obesity. Probiotics' health-promoting properties include gut homeostasis maintenance, pathogen alienation, nutrient bioavailability enhancement, and stimulation and modulation of the host immune system11. Probiotics are currently the primary focus of investigation as potential biotherapeutics for treating numerous gastrointestinal diseases, liver damage, malignancies, and inflammatory and metabolic disorders12,13.

 

A high-calorie diet that causes gut dysbiosis substantially reduces the diversity of bacteria, enhances calorie absorption, disrupts intestinal barrier integrity, and causes chronic inflammation that leads to metabolic disorders: endotoxemia, production of reactive oxygen species, and reduced regulation of genes involved in metabolic processes. All of these factors contribute significantly to the development of metabolic complications14. A new bio intervention utilizing probiotics, prebiotics, and synbiotics that may restore intestinal homeostasis must be created to prevent metabolic diseases.

 

Various probiotics help treat this health condition, raising interest in developing probiotic treatments. Probiotic therapy may be an additional option for improving gut microbiota composition15. Obesity and obesity-related disorders are impacted by gut microbiota composition as well as metabolite changes16–18. Commercial strains are more likely to contain probiotic strains from the Lactobacillus and Bifidobacterium genera19. An experimental study on L. fermentum CECT5716 demonstrated its ability to modify the microbiome to improve obesity and understanding the pathophysiology of obesity20. Several beneficial microbes, including Bifidobacterium and Lactobacillus spp., have been shown to decrease obesity, fatty liver, and chronic inflammatory disorders21.

 

The primary lactic acid bacterium detected in dadih produced by water buffalo in the Tanjung Bonai area of Tanah Datar, West Sumatra, is Lactoplantibacilus pentosus. The greatest results were obtained with fermented milk products containing 20% sweet orange juice and 6% starter L. pentosus. The total LAB colonies ranged from 4.67×10^9 to 9.0×10^9 CFU/mL22, with antioxidant activity varying between 25.04% and 37.71% and total phenol content between 38.32 and 67.20 mg GAE/gr23. Due to strain variations, it is unknown whether L. pentosus can effectively prevent obesity or reduce inflammation; additionally, its molecular mechanisms must be investigated. L. pentosus will be explored for its preventative effects on lipid synthesis and anti-inflammatory properties, making it a viable nutraceutical or nutritional supplement.

 

MATERIALS AND METHODS:

Materials:

L. pentosus was found in Dadih, a traditional fermented dairy product from Lintau Buo Utara, Tanah Datar, West Sumatera (Indonesia), which contains Lactobacillus strains with possible probiotic properties. It was grown at 37°C in MRS broth, incubated overnight, and harvested by centrifugation at 3000g and four °C for 15 minutes. The probiotic strain was suspended in sterile saline and fed to mice through oral ingestion. Preparation of the milk product fermented by Lactobacillus pentosus was prepared as described previously (23). Every week, fermented milk is made and given to mice 1 mL daily (Lactobacillus pentosus count: 109CFU/mL).

 

Animal Groups and Feeding:

The research animals were authorized by the Ethics Committee of the FK Unand (Approval No. 346/UN.16.2/KEP_FK/2023). To calculate the necessary number of samples, Federer's formula was applied, requiring more than six subjects. We used twenty-eight male Wistar rats weighing between 150-180 grams from the Faculty of Pharmacy at Andalas University, divided into four groups with seven rats in each. The conditions for raising the rats included a constant temperature of 22 ±2 degrees Celsius, 55±10% humidity, and alternating 12-hour light/dark cycles. Initially, all rats were fed a standard diet for a week to acclimatize. After this period, they were split into four different dietary groups (each containing seven rats): a normal diet (ND), a high-fat diet (HFD), a high-fat diet supplemented with Lactobacillus pentosus 109 CFU/m (HFDL) and HFDSF (High Fat Diet supplemented with fermented milk containing L. pentosus at 109 CFU/ml). The normal diet comprised 4.0kcal/g, with 20% of calories derived from protein, 12% from fat, and 68% from carbohydrates. Meanwhile, the high-fat diet provided 4.7kcal/g, with 16% of the calories coming from protein, 42% from fat, and 43% from carbohydrates.

 

Table 1. Animal Group and Feeding

Group

Treatment

ND ( Normal diet/control)

-

HFD (High fat diet)

-

HFDL (high fat diet_L.pentosus)

L.pentosus 109 CFU/mL per day

HFDFL (high-fat diet fermented milk L.pentosus)

1 ml Fermented milk and L.pentosus 109 CFU/mL/day

 

The body weight was recorded weekly using an appropriate scale (ACIS Digital Compact Balance BC Series). After seven weeks, the rats were euthanized. Mice were starved for 8 hours and given anaesthesia with diethyl ether before sacrifice. After an eight-hour fast, the mice's blood was collected. The mice's liver, adipose tissues, and mesenteric tissues were obtained shortly after euthanising. The tissues were washed with phosphate-buffered saline.

 

Measurement of superoxide dismutase (SOD):

After collecting blood, it was centrifuged and then transferred to an EDTA tube. To measure the levels of Superoxide Dismutase (SOD) in the plasma, we utilized the SOD Activity Kit (232-943-0, Merck, Sigma-Aldrich, Darmstadt, Germany) following the ELISA method. We added xanthine oxidase reagent to the plasma, then placed the mixture in a 1 mL glass cuvette and let it rest at room temperature for 30 minutes. During this time, the xanthine oxidase generated superoxide, which is detectable at a wavelength of 560 nm.

 

Measurement of malondialdehyde (MDA):

The protein-bound malondialdehyde (MDA) was measured using the MDA Assay Kit (competitive ELISA) (ab238537). Each serum tube received 2.5ml of 5% trichloroacetic acid (TCA), and the mixture was homogenized with a vortex mixer before being centrifuged at 10,000rpm for 15 minutes. Subsequently, 1mL of thiobarbituric acid (TBA) reagent was added to each tube, which were then incubated in a water bath at 100 degrees Celsius for 30 minutes. After cooling, the results were measured with a spectrophotometer at 530 nm.

                          Sample Absorbance

MDA levelsl = ---------------------------- X C. Standard

                           Standard absorbent

 

Liver histological analysis:

The liver specimens were preserved in 10% formaldehyde for a day, embedded in paraffin, and then sectioned into 3-micrometer slices using a microtome. These sections were subsequently deparaffinized and stained with Hematoxylin and Eosin (H&E). Lipid accumulation and hepatocyte inflammation were investigated using an optical microscope (Olympus CX33, Tokyo, Japan) at 100 and 400 magnifications24

 

An optical microscope at 100x (objective 10x) and 400x (objective 40x) magnification (Olympus CX33, Tokyo, Japan) and adipocyte cell diameter were measured using a Sony Exmor Beta camera 3.14MP. In addition, the beta view program was used at 400x magnification to measure a minimum of 50 adipocyte cells, which were then displayed as a mean value (µm). (24)

 

The scoring system criteria for liver histology assessment are based on Veteläinen RL25

 

The scores for hepatic steatosis are as follows:
0 = The proportion of fat in all hepatocytes is less than 10%
1 = Fat is present in 10–30% of all hepatocytes.
2 = 31-60% of the hepatocytes in total are adipose

3 = Over 60% of all hepatocytes are composed of fat.

 

The scores for lobular inflammation are as follows:

1 = None

2 = For every 200x field, there are fewer than two sites of inflammation.

3 = two to four sites of inflammation per 200x field

4 = more than four foci of inflammation per 200x field.

 

Hepatic balloning is assessed as:

0 = None

1 = indicates that there is minimal ballooning.

2 = represents prominent balloons.

 

Statistical Methods:

For analysis, the SPSS statistical program (version 23.0) was utilized. The data are reported as mean and standard deviation. We applied a one-way ANOVA with a posthoc LSD test to evaluate differences in body weight, serum SOD, and MDA levels among groups. A p-value of less than 0.05 was used to classify differences as statistically significant.

 

RESULT:

Effect of L. pentosus on Body Weight:

The initial body weights of the mice were consistent across all four groups, ranging from 230 to 243 grams, as illustrated in Figure 1. Throughout the rearing period, there was a general increase in the body weights of all groups. The normal diet/control (ND) group had the lowest body weight, while the High Fat Diet (HFD) group had the highest. Following a 6-week high-fat diet, the HFD group gained 24.30% more weight. In comparison, the body weight of the mice in the High Fat Diet with Lactobacillus pentosus (HFDL) group (23.78 ±0.84g) was significantly lighter than that of the HFD group (26.59±1.17 g) with a p-value of less than 0.05.

 

Figure 1. Changes in body weight during seven weeks

 

Levels of superoxide dismutase (SOD):

The analysis of Figure 2 revealed a significant difference in Superoxide Dismutase (SOD) levels between the control group and the High Fat Diet (HFD) group, as determined by ANOVA with Tukey's posthoc test (p<0.05). Furthermore, the administration of L. pentosus in the High Fat Diet (HFDL) group resulted in statistically significant reductions in SOD levels compared to the control group (p<0.05). Our findings indicate that animals treated with a High Fat Diet (HFD) supplemented with Lactoplantibacilus pentosus and fermented milk containing Lactoplantibacilus pentosus experienced a significant decrease in Superoxide Dismutase (SOD) levels compared to the control group (p < 0.05). Conversely, SOD levels in the HFD group that received Lactoplantibacilus pentosus alone and in combination with fermented milk were significantly higher than those in the HFD group, as depicted in Figure 2 (p< 0.05).

 

Figure 2. Comparison of SOD levels between groups

 

Malondialdehyde (MDA) levels:

There were significant changes in malondialdehyde (MDA) levels between the control, High Fat Diet (HFD), High Fat Diet with Lactoplantibacilus pentosus (HFDL), and High Fat Diet with fermented milk and Lactoplantibacilus pentosus (HFDFL) groups (p<0.05), as shown in Figure 3. MDA levels were highest in the HFD group, with rats on this diet showing a significant increase in serum MDA levels compared to the control group (p<0.05). In contrast, MDA levels were significantly reduced in the HFDL and HFDFL groups compared to the HFD group, with values of 1.32±0.04, 1.38±0.03, and 1.52±0.04 nmol/mg protein, respectively (p=0.002).

 

Figure 3. Comparison of MDA levels between groups

 

Adipocyte Histological Image of Mice:

Figure 4 shows that the animals with high-fat diets showed hypertrophy in adipocyte cells, which was more significant than in control cells. The mean adipocyte diameter is 67,72 and 44, 5µm, respectively (Table 2). Adipocyte hypertrophy symptoms were reduced after treatment with fermented milk and Lactobacillus pentosus isolate; the mean adipocyte diameters were 50,38 and 54,81µm


Negetive Control

High fat diet

Orange fermented milk

Lactobacillus pentosus Isolate

Figure 4: Adipocyte cells (A) and a small amount of thin connective tissue containing a vascular (V) are visible in the adipose tissue histology from experimental animals. Negative control group (a, e), high-fat diet (b, f), orange fermented milk (c, g), and lactobacillus isolate treatment (d, h): Hematoxylin and eosin stains, with the top panel at 10x magnification and the bottom panel at 40x.

 


Table 2. Histopathological mean of adipocyte diameter in all groups

No

Parameter histologies

Mean of adipocyte diameter (µm)

Sample

Mean

Mean group

1

Negative Control

Fat K (-) neg 2

43,55

44,55

 

 

Fat K (-) neg 4

45,54

2

High-fat diet

Fat K (+) HFD 5

79,06

67,72

 

 

Fat K (+) HFD 6

56,38

3

Fermented milk

Fat P 1 Fermented milk 5

53,71

50,38

 

 

Fat P 1 Fermented milk 7

47,05

4

Lactobacillus isolate

Fat P Lactobacillus isolate 1

54,86

54,81

 

 

Fat P Lactobacillus isolate 7

54,76

 

 

Liver Histological Image of Mice:

Figure 5 shows hepatocytes grouped in order and establishing an average histologic impression in control animals. Animals with a high-fat diet induce specific hepatocytes with intracytoplasmic clear vacuoles (↓), inflammatory cells dispersed throughout the body (), and areas of moderate fibrosis, particularly periportal (↓↓). The hepatic steatosis score in the HFD group was 2.1 (30%-60% steatosis), the inflammation score was 2.1 (less than two inflammatory foci per 200 x feld0), and the necrotic and fibrotic score was 1 (10-30%). Meanwhile, the liver histology score in the normal group was the lowest (Table 3). There was a decrease in steatosis, inflammation, necrotic, and fibrotic scores in the HFD group given fermented milk with Lactobacillus pentosus. Lipomatosis, inflammatory cell infiltration, and fibrosis were reduced after fermented milk and Lactobacillus pentosus isolate treatment.


 

Negetive Control

High fat diet

Orange fermented milk

Lactobacillus pentosus Isolate

Figure 5. shows the histology of experimental animals' liver tissue, which includes the hepatic parenchyma, central vein (V), port area (P), and hepatocyte cells (H) grouped in trabeculae with sinusoids. Negative control group (a, e), high-fat diet intervention (b, f), orange fermented milk administration (c, g), and Lactobacillus isolate treatment (d, h). The lower panel has a 40x objective, and the upper panel has a 10x objective in hematoxylin-eosin.

 

Table 3. Histopathological scores in all groups

No

Histologic parameter

Liver histologic score

Steatosis

Inflammation

Nekrotic

Fibrosis

Mean sample

Mean grup

Mean sample

Mean grup

Mean sample

Mean grup

Mean sample

Mean grup

1

Negative Control

Fat K (-) neg 2

0,2

 

0,0

 

0,0

 

0,0

 

 

 

Fat K (-) neg 4

0,2

0,2

0,2

0,1

0,0

0,0

0,0

0,0

2

High-fat diet

Fat K (+) HFD 5

2,2

 

2,0

 

1,0

 

0,6

 

 

 

Fat K (+) HFD 6

2,0

2,1

2,2

2,1

1,0

1,0

0,8

0,7

3

Fermented milk with Lactobacilus

Fat P 1 Fermented milk 5

1,0

 

0,8

 

1,0

 

0,0

 

 

 

Fat P 1 Fermented milk 7

1,2

1,1

0,8

0,8

1,0

1,0

0,2

0,1

4

Lactobacilus pentosus isolat

Fat P Lactobacilus isolate 1

1,0

 

0,8

 

1,0

 

0,2

 

 

 

Fat P Lactobacillus isolate 7

1,0

1,0

1,2

1,0

0,6

0,8

0,2

0,2

 


DISCUSSION:

Obesity is the result of a combination of internal and external factors. Genetic makeup and environmental factors influence intake and energy imbalances, changes in gut microbiota, and inappropriate eating patterns. Obesity is a significant and severe health issue in both developing and developed countries. Leptin signalling is essential in controlling both satiety and body weight26. Ghrelin and Leptin are two hormones that play an essential role in BMI regulation27. Type 2 diabetic patients had significantly higher serum leptin levels28. Healthy food choices, regular physical activity, and behaviour modification to maintain lifestyle changes are all critical factors in preventing obesity and living a healthy life29.

 

Numerous studies have explored the modification of the gut microbiota through the administration of probiotics in various settings, including in vitro experiments, in vivo animal models, and human clinical trials, aiming to address obesity. Probiotics can thrive and colonize the surface of the intestines, providing benefits to the host when their populations reach effective levels30. Giving probiotics to prevent obesity has recently become a worldwide concern. Providing or combining one probiotic strain with a different prebiotic strain will have a different effect. Probiotics are regarded as one of the most important methods for modifying the gut microbiota composition, which can influence food intake, metabolic activity, and body weight composition31. Our previous study found that the antioxidant activity of fermented milk Lactoplanti bacillus pentosus and the addition of orange varied from 25.04% to 37.71%. Additionally, the total phenolic content in these samples ranged from 38.32 to 67.20 mg GAE/gr23. This study tested the probiotic Lactoplantibacilus pentosus derived from dadih (a traditional Minangkabau food) for its prophylactic potential in rats induced by a high-fat diet. Antioxidants are essential for preventing the formation of reactive oxygen and nitrogen species, which can cause damage to DNA, lipids, proteins, and other biomolecules. These reactive species are known to contribute to cellular damage and various diseases. Antioxidants protect tissues from free radical damage32. Both oxidative stress and inflammation are linked to endothelial dysfunction and reactive oxygen species (ROS), a byproduct of lipid and protein oxidation33.

 

In this study, the intervention group gained weight after consuming a high-fat diet for a period of six weeks. Meanwhile, a daily intragastric administration of L. pentosus at 109 CFU/mL for six weeks can suppress weight gain compared to the HFD group. Several studies have found a significant impact by modifying the composition of the microbiota. The probiotic Lactobacillus rhamnosus strain LRH05 was administered to high-fat-fed mice and showed specific effects in ameliorating pro-inflammatory processes by lowering inflammatory markers. As a result, LRH05 can be regarded as a potential probiotic strain for obesity prevention34. Another study discovered that Lactobacillus paracasei L9 supplementation reduced dislipidemia, fatty liver and inflammation in HFD-induced mice. Lactobacillus paracasei L9 supplementation may be a natural way to reduce obesity35. The addition of the synbiotic L. pentosus GSSK2+isomalto-oligosaccharide significantly prevented weight gain, decreased abdominal size, reduced the Lee index, lowered BMI, and minimized visceral fat accumulation in Sprague Dawley rats on a high-fat diet for 12 weeks36.

 

The malondialdehyde (MDA) levels in the HFD group significantly increased compared to the normal group, while the levels of Superoxide Dismutase (SOD) significantly decreased. These findings align with human research that reported significantly higher MDA levels in obese individuals compared to healthy controls. Additionally, antioxidant molecules and enzymes, including Glutathione (GSH) significantly lower in both the obese and obese diabetic groups37. Free radicals damage the liver and kidneys. Lead exposure in rats significantly elevated malondialdehyde (MDA) levels while simultaneously reducing the activities of superoxide dismutase (SOD) and catalase, as well as the concentrations of glutathione (GSH) in the liver and kidneys38. Conversely, animals that were administered probiotics exhibited higher levels of the antioxidant enzyme SOD and lower levels of MDA compared to those in the High Fat Diet (HFD) group. Khana et al. 2021 recently demonstrated that oral administration of a probiotic mixture of L. pentosus GSSK2 + isomalto-oligosaccharides effectively alleviated hepatic oxidative stress inmice fed a high fat diet (HFD). This intervention significantly increased Superoxide Dismutase (SOD) levels while decreasing malondialdehyde (MDA) levels. Furthermore, it was observed that MDA levels were greater in the HFD group than in the control group. MDA content increases due to increased lipid peroxidation in the cytomembrane, which damages the cytomembrane. Malondialdehyde is an important lipid peroxidation product that can be used to monitor cytomembrane lipid peroxidation. SOD and other cellular enzymes can be used to detect cell defence responses to external stress. To protect against external injury, the activity of enzymes such as SOD within cells increases reactive oxygen removal and a reduction in oxidative damage 39. SOD levels can be increased by taking probiotics. This is in line with Lactiplantibacillus pentosus CQZC01 supplementation, which increased superoxide dismutase (SOD) levels while decreasing malondialdehyde (MDA) content, thereby increasing antioxidant capacity. Furthermore, by increasing the levels of this factor, Lactiplantibacillus pentosus CQZC01 can alleviate the damage caused by alcohol to the gastric mucosa: gastric mucosal defence, oxidative stress inhibition, and inflammation suppression40. The antioxidant mechanism of probiotics can be through free radical scavenging, metal ion chelation, enzyme regulation, and intestinal microbiota modulation41.

 

Although the specific function of oxidative stress in the progression of obesity is unknown, it is commonly acknowledged that cellular oxidative damage might contribute to different metabolic dysfunctions associated with obesity.Several studies have found that oxidative stress is linked to increased body fat mass via adipogenesis and lipogenesis42. Obesity has been linked to oxidative stress by increasing preadipocyte proliferation and adipocyte differentiation43. On the other hand, adiposity causes increased oxidative stress through several biochemical processes, such as superoxide formation44. Chronic oxidative stress and associated inflammation in obese individuals are linked to the disruption of cellular signaling and metabolism, leading to insulin resistance, metabolic dysfunction, diabetes, and cardiovascular disease.45. Pharmaceutical formulations based on antioxidants are widely used to prevent and treat free radical-related diseases such as atherosclerosis, stroke, diabetes, Alzheimer's disease, and cancer46.

 

Research indicates that fermented foods can positively impact gut health and metabolism in several ways. They enhance the growth of short-chain fatty acid (SCFA)-producing bacteria and adjust the ratio of Firmicutes to Bacteroidetes, which is often linked to healthier metabolic profiles. Additionally, fermented foods can reduce intestinal permeability (often referred to as "leaky gut"), decrease inflammation and oxidative stress, and boost lipid metabolism47. These benefits make fermented foods a valuable component of diets aimed at improving overall health and managing conditions like obesity and metabolic syndrome. LAB supplementation can help to regulate the gut microbiota. Changes in diet and gut microbiota are strongly associated with oxidative stress. In mice fed a high-fat diet, a decrease in MDA and an increase in total antioxidant capacity showed a strong positive relationship with Lactobacillus48. Adipocyte hypertrophy occurred in animals fed a high-fat diet, and symptoms of adipocyte hypertrophy were alleviated by administering fermented milk and Lactobacillus pentosus isolate. Previous research on mice induced by high fat yielded an anti-obesity effect via modulating inflammation, downregulating lipogenic genes, and decreasing the number of adipocytes49. Single probiotics based on Lactobacillus reduce visceral and subcutaneous adipose tissue, whereas probiotics based on Bifidobacterium only reduce visceral adipose tissue50. Lactobacillus pentosus strain S-PT84 supplementation inhibited the development of hepatic inflammation and fibrosis51. The protective effect of this L. pentosus from dadih is comparable to that of L. pentosus CQZC0, Lactobacillus delbrueckii subsp.Bulgarian52. Lactobacillus pentosus derived from dadih may have a suitable hepatoprotective action for people who frequently consume high-fat diets.

 

CONCLUSION:

Fermented milk Lactiplantibacillus pentosus was discovered to reduce fat accumulation in high-fat mice. Animals treated with Lactiplantibacillus pentosus had higher SOD levels and lower MDA levels than the HFD group. Adipocyte cell hypertrophy was more pronounced in animals fed a high-fat diet than in a control diet. Meanwhile, administering fermented milk and Lactiplantibacillus pentosus alleviated the symptoms of adipocyte hypertrophy. The HFD group had higher steatosis, inflammation, fibrotic, and necrotic liver scores than the normal diet group. Steatosis, inflammation, necrosis, and fibrotic scores decreased in the HFD group given fermented milk containing Lactiplantibacillus pentosus. Lipomatosis, inflammatory cell infiltration, and fibrosis were all reduced following the administration of fermented milk and Lactiplantibacillus pentosus. Lactiplantibacillus pentosus from dadih has been shown to reduce obesity, hepatic steatosis, and systemic inflammation in people who consume high-fat diets regularly.

 

CONFLICT OF INTEREST:

The authors declared that there is no conflict of interest.

 

ACKNOWLEDGMENTS:

Under the research implementation contract for Riset Publikasi Bereputasi (RPB) No. 106/SPK/PTN-BH/FKep/Unand-2023, Dr. dr. Susmiati, M. Biomed, this study sponsored the LPPM Universitas Andalas. For permission and facilities used during the research, the authors are grateful to the Laboratory of Animal Product Technology Faculty of Animal Husbandry and Faculty of Pharmacy Andalas University, Padang.

 

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Received on 17.12.2023      Revised on 14.05.2024

Accepted on 21.08.2024      Published on 20.01.2025

Available online from January 27, 2025

Research J. Pharmacy and Technology. 2025;18(1):94-102.

DOI: 10.52711/0974-360X.2025.00015

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