INTRODUCTION:

Numerous lactic acid bacteria (LAB) found in fermented foods or the human microbiome have been proved to benefit human health. Probiotic LAB strains have been demonstrated in recent studies to boost both the innate immune system and the acquired immune system, avoiding immunological diseases and offering protection against bacterial and viral infection.¹ Therefore, it is necessary to examine the ability of P. acidilactici to produce adipokines and others that are capable of being TNF inhibitors. Expression is seen in silico using a bioinformatics approach to this secondary metabolite of Pediococcus acidilactici DS1.

There are around 600 adipokines known, but the ones that have been investigated the most include leptin, adiponectin, resistin, visfatin, tissue factor, tumour necrosis factor alpha (TNF-), transforming growth factor beta, monocyte chemoattractant protein-1 (MCP-1), interleukin 6 (IL-6), and IL-8. Given the pleiotropic significance of leptin in energy balance, the therapy of malnutrition by the introduction of commensal bacteria able to control its production and function may be an appealing approach, but the right strains must be chosen.²

Inflammation is brought on by a protein in your body called tumour necrosis factor. TNF inhibitors are medications that reduce inflammation and are used to treat inflammatory diseases such psoriasis, rheumatoid arthritis, juvenile arthritis, inflammatory bowel disease, Crohn's disease, and ulcerative colitis. Targeting a chemical known as tumour necrosis factor that causes inflammation, they decrease inflammation and potentially halt the course of illness (TNF). Previous research explained that probiotics contain many benefits, one of which is anti-inflammatory, immunomodulatory and even anti-diabetic. This will certainly be beneficial for human health so that they can take advantage of local biodiversity as prevention and even treatment. especially on TNF inhibitors because of the research that has been done probiotics are capable of being TNF expression inhibitors.

The probiotic bacteria, commonly referred to as "gut warriors," are dietary components with therapeutic potential that fight pathogenic bacteria by preventing them from colonising the gut epithelium and increasing the establishment of beneficial gut microbiota.^3,4 Probiotic microorganisms have undergone substantial research for medicinal uses in conditions including IBS, antibiotic-associated diarrhoea, systemic allergies, and age-related illnesses.^5,6 Probiotics are also known to have the ability to restore the beneficial gut microflora that have been lost as a result of medication treatment.⁷ Probiotics' is a functional food that contains a living microbe that, when given to the host in sufficient quantities, has a beneficial effect on the host's health^8-14. This study aims to determine the results of secondary metabolites in Pediococcus acidilactici DS1 on TNF inhibitor expression and as an immunomodulator.

MATERIAL AND METHODS:

Bacteria used were Pediococcus acidilactici DS1 culture isolated from Dadiah Bukittinggi collection from the Animal Products Technology Laboratory, Andalas University.

Search of Secondary Metabolite Search:

Search for bacterial secondary metabolites was carried out by studying literature from published journals. Each compound that has been determined is then searched for its canonical structure and isomeric SMILE (simplified molecular-input line-entry system) in the PubChem database (https://pubchem.ncbi.nlm.nih.gov/).

Prediction of bacterial content's Structure-activity Relationship (SAR):

Secondary metabolites in bacteria from the literature study were analyzed for their potential using WAY2DRUG PASS prediction (http://www.pharmaexpert.ru/passonline/predict.php) as a cholesterol-lowering treatment. Previously, each compound needed to find the SMILE structure obtained from the Pub Chem database. Then the potency of the compound was analyzed using WAY2DRUG PASS prediction to determine its potential for cholesterol reduction treatment. We recommend using a score of 0.5 as the cut off score. The Pa value means the accuracy of the prediction function obtained, the higher the Pa value of a function, the better the accuracy level.¹⁵

RESULTS AND DISCUSSION:

Result Prediction QSAR Way 2 Drug Pass Server from Pediococcus acidilactici DS1 from dadiah Sianok in table 1.

Table 1: Result Prediction QSAR Way Drug Pass Server from Pediococcus acidilactici DS1

Compound	TNF Expression Inhibitor	Immunomodulator
Butyric Acid	0.637	0.407
Propionic Acid	0.577	0.365
Lactic Acid	0.638	0.334
Valeric Acid	0.646	0.504
Caproic Acid	0.646	0.504
Average	0.63	0.42

The capacity of probiotics to interact with epithelium, dendritic cells, macrophages, and lymphocytes makes them potential immunomodulators. Dendritic cells in the digestive system may identify the parts of the L. lactis bacterial wall and can alter how they operate. Additionally, probiotic LAB might cause an inflammatory reaction.^16,17 Intestinal microbiota will drive macrophages and dendritic cells to release IL-6, IL-1, and TNF, which will encourage Th17 differentiation. This impact can be observed by the rise in TNF- in dadih-induced animals compared to the control group. Th17 contributes to the release of cytokines that promote inflammation, such IL-17. IL-6, IL-12, and TNF-a production may be increased by certain probiotic bacteria. Probiotics were shown to be able to stimulate the production of the cytokine IL-10, which is produced by a variety of cells, including Th-2, macrophages, dendritic cells, keratinocytes, and Treg cells. Th-1.6 response is suppressed by IL-10, which has anti-inflammatory properties. In this experiment, mice that had been given dadih had greater levels of the cytokine IL-10 than mice that hadn't received it. Probiotics may stimulate Foxp3+ Treg cells, which generate TGF- and IL-10, according to certain research. Probiotics influence adaptive immune cells in the digestive tract to create pro-inflammatory and anti-inflammatory cytokines via interactions with enterocytes, dendritic cells, Th1, Th2, and Treg. Result of Heat Map Potensi QSAR Metabolite Pediococcus acidilactici DS1in figure 1.

Figure 1: Heat Map Potention QSAR Metabolite Pediococcus acidilactici DS1.

If the Pa value is more than 0.7, it indicates that the compound is predicted to have a high potential as an anti-inflammatory (eg) because of the high similarity with the compound in the database that has been proven as a treatment. On the results of the QSAR Way drug prediction, it can be seen that the secondary metabolite Pedioccus acidilactici as an immunomodulator has a value of 0.42 which is at the midpoint with a value half of the highest value. On the heat map, there is a yellow color that shows high and better results, we see on the immunomodulator the color that is produced is purple, meaning that Pediococcus acidilactici from Dadiah Bukittinggi can be treated as a therapy to increase immunoglobulins.

Pediococcus metabolites have a high potential as an anti-inflammatory (0.55). This score is supported by the potency of dadiah as a TNF expression inhibitor (0.63), Histamine Release Inhibitor (0.58), and JAK2 expression inhibitor (0.55). TNF (Tumor Necrosis Factor) is one of the pro-inflammatory cytokines that can trigger inflammation. TNF plays a crucial role in the regulation of innate immune responses. TNF triggers the secretion of inflammatory mediators and is involved in the activation and function of innate immune cells, TNF is also involved in the proliferation, differentiation and survival of macrophages.¹⁸ TNF-alpha is one of the pro-inflammatory mediators involved in the pathogenesis of T2DM.¹⁹ High histamine levels have been reported to be associated with diabetic long-term complications.²⁰The value of Pa (Probability To Be Active) is a value that describes the potential of a compound being tested. Determination of this value is done by comparing the structure of the input compound with a compound that has been proven as a particular treatment.

Figure 2: Schematic of NF-Kappa B activation by TNF stimulus

NF-κB is a family of dimeric transcription factors consisting of 5 proteins, namely p65 or RelA, RelB, c-Rel, p50 and p52.²¹ NF-B activation can be stimulated by TNFα molecules. TNF will bind with TNF receptors. Then signal transduction occurs when the TNF receptor has been activated. Due to the TNF-TNFR interaction then IκB undergoes IκB phosphorylation. Phosphorylation of IκB leads to degradation. When IκB is degraded, NF-B dimers (eg, p65/p50 or p50/p50 subunit combinations are possible) and translocates to the nucleus, where it binds by consensus to the DNA sequence of the target DNA and activates gene expression such as genes involved in pro-inflammation including TNFα.²²

CONCLUSION:

Based on the approach through Prediction QSAR Way Drug Pass Server and Heat Map Potential QSAR Metabolite on Pediococcus acidilactici DS1 bacteria isolate from Dadiah Bukittinggi, it shows that this bacterium can be applied as therapy for TNF expression inhibitors and as immunomodulators. Suggestions for future research can be carried out in vivo on experimental animals to see the effectiveness of these bacteria.

ACKNOWLEDGMENT:

This Research project is supported by Baiturrahmah University and Andalas University.

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Received on 30.09.2022 Modified on 27.01.2023

Research J. Pharm. and Tech 2023; 16(10):4737-4740.

DOI: 10.52711/0974-360X.2023.00769