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Leukotriene Receptor Antagonist Montelukast Shows Therapeutic Promise Against Non-Alcoholic Steatohepatitis

Non-alcoholic steatohepatitis (NASH) is a kind of liver inflammation and damage resulting from fat accumulation in the liver. It's an umbrella term for liver disorders that affect people who drink little to no alcohol but are diagnosed with a "fatty liver." Nonalcoholic fatty liver disease (NAFLD) patients often suffer from no symptoms, but once NAFLD promotes inflammation and destroys liver cells, making the liver malfunction, the condition has developed into a more aggressive form of fatty liver disease, NASH, which can scar the liver or even lead to cirrhosis and liver cancer. Studies have found that dyslipidemia, particularly hypertriglyceridemia, oxidative stress, and insulin resistance exert a key influence on the development of NASH. Moreover, the production of different atherogenic factors, including pro-inflammatory cytokines, is also considered to be related to the development of NAHS. However, currently, there is no specific treatment strategy for NASH, though some approaches, including insulin-sensitizers, lipid-lowering drugs, antioxidants, L-carnitine, and probiotics, have been proposed to treat NAFLD.

 On this condition, researchers from Egypt have recently published an article investigating the efficacy and safety of the leukotriene receptor antagonist montelukast in treating NASH. Cysteinyl leukotrienes (CysLTs) are generated by inflammatory cells that contain LTC4 synthases, such as eosinophils, basophils, mast cells, and erythrocyte endothelial cells, while the liver is essential for leukotriene production and metabolism (LTs). There is a bunch of research evidence that leukotriene plays a significant role in obesity, insulin resistance, and fatty liver disease, and leukotriene produced by the 5-lipoxygenase pathway is important in inflammation and chemotaxis.

 Based on these studies, researchers further noted that montelukast, an approved CysLT1 receptor antagonist for the maintenance treatment of asthma and to relieve symptoms of seasonal allergies, can block the pro-inflammatory action of leukotriene D4 (LTD4). Furthermore, montelukast can ease insulin resistance, an important part of the pathogenesis of NAFLD. And later research has demonstrated that montelukast can protect against hepatotoxicity caused by acetaminophen in a rat model.

 These study results encourage researchers from Egypt to evaluate the efficacy and safety of montelukast as a potential therapy to treat patients with NASH. Experiments by this research team aim at evaluating the effect of montelukast on liver stiffness, serum levels of liver fibrosis biomarkers (hyaluronic acid (HA) and transforming growth factor beta1 (TGF-β1), insulin resistance, and metabolic and inflammatory parameters.

 By recruiting 52 qualified patients from the National Liver Institute, Menoufia University, Egypt, scientists screened and measured the efficacy of montelukast on liver stiffness, liver panel, serum levels of liver fibrosis biomarkers (HA and TGF-β1), insulin resistance, metabolic and inflammatory parameters.

 It turned out that montelukast has a promising therapeutic effect on overweight and obese patients with non-alcoholic steatohepatitis by improving liver stiffness, hepatic fibrosis biomarkers, and metabolic parameters, as well as significantly decreasing the inflammatory and oxidative stress biomarkers in these NASH patients. In conclusion, the leukotriene receptor antagonist montelukast may be a potential therapy for non-alcoholic steatohepatitis patients with good tolerance and without provoking depression compared with existing treatments.

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Creative Biolabs has established several approaches utilized for drug loading into exosomes .

After years of hard-working in exploring the application of exosomes, Creative Biolabs has got all the necessary experience and technologies of exosome engineering. The most usually used approaches are loading naïve exosomes isolated from parental cells ex Vitro, loading parental cells with a drug, and transfecting/infecting parental cells with DNA encoding therapeutically active compounds. We will respectively give a brief introduction to each of them.

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Drug Loading into exosome

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Creative Biolabs offers a full range of antigen and antibody conjugation services to meet our global client’s specific requirements.

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Creative Biolabs offers various technologies applied for exosome characterization, including nanoparticle tracking analysis, FACS sorting, w

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Creative Biolabs provides customized IVD antibody development services targeting different cancer biomarkers to global clients.

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Creative Biolabs has advanced platforms for high-precision exosome proteomics services to study various protein products and other contents

It is reported that exosomal proteins have an essential role in the pathological process of diseases. Therefore, it is necessary to analyze and identify exosomal proteins. Advanced progress in mass spectrometry (MS)-based proteomic analyses of mammalian exosomes derived from different cell types and body fluids have led to the identification of thousand vesicular proteins. With the help of Creative Biolabs, an expert in exosome technical services, essential clues can be provided for customers to decipher molecular mechanisms involved in vesicle cargo sorting, biogenesis and protein functions of exosomes.

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Creative Biolabs is very proud of offering a full range of exosomal antibodies that can be used for exosome marker detection via various tec

Exosome Antibody

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Creative Biolabs offers DNA sequencing services for our global customers to advance the development of IVD.

The sequence of nucleic acids in polynucleotide chains ultimately contains the information for the hereditary and biochemical properties of terrestrial life, which has been an important reference for in vitro diagnostics (IVD). Therefore, the ability to measure or infer DNA sequences is imperative to biological research, especially in the IVD field. With our deeply-rooted IVD expertise and abundant experience, Creative Biolabs provides DNA sequencing services to accelerate the development of your IVD projects.

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The capacity of exosomes to employ as biovesicles for proteins, lipids, and nucleic acids, and their role in intercellular communication, make them a powerful platform for biomarker discovery and drug delivery. Therapeutically, exosomes have been used in a variety of forms. They are inherently bioactive so that can be isolated from cells for downstream use without further modification.

Creative Biolabs provides varieties of high-quality exosome engineering services for academic research and clinical practices.

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Creative Biolabs provides a comprehensive range of IVD kit and assay development services.

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Creative Biolabs provides comprehensive exosome-related services to global clients.

With years of experience in the field of molecular and cell biology research, Creative Biolabs is capable of supporting all aspects of your exosome research. Equipped with state-of-the-art tools and technologies, we offer the most comprehensive exosome-related research services, ranging from isolation and characterization to cargo profiling and engineering, for a better understanding of their physiological functions, for the discovery of clinical biomarkers, and engineering of exosomes for therapeutic applications.

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Creative Biolabs has developed validated protocols to purify exosomes from both plasma and other body fluids with decreased co-purification

Exosomes, which are freely available in plasma and other body fluids, have attracted a great deal of interest as therapeutic and diagnostic biomarkers. With years of experiences in exosome technology, Creative Biolabs has developed validated protocols to purify exosomes from both plasma and other body fluids with decreased co-purification of non-exosomal proteins.

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How to Prepare and Construct an RNA Sequencing Library?

RNA is a polymeric molecules composed of nucleotides that are required for gene coding, decoding, regulation, and expression. In molecular biology, DNA's genetic information gets transcribed into multiple copies of messenger RNA (mRNA), and then mRNA is translated into proteins. This is why cells can synthesize several protein molecules from a single gene at a given point of time. However, only 1-4% of total RNA are mRNA, the rest of which are generally non-coding, called non-coding RNA (ncRNA). ncRNA is unable to be translated into protein but there are many types of them, which have different biological functions.

Therefore, it's necessary to analyze the total RNA molecules for a better understanding of gene expression, which is called transcriptome analysis. Gene expression analysis can be used to monitor the response of a gene to treatment with a compound or drug of interest. Leveraging features of the next-generation sequencing (NGS), RNA sequencing (RNA-Seq) can efficiently detect and quantify RNA in a biological sample at a given point in time. Researchers often take advantage of the high-throughput sequencing data, broad dynamic range, and high sensitivity from NGS to investigate the whole transcriptome of a single cell or a population of cells without using probes and to detect differentially expressed genes.

Protocol of RNA-seq

Though with a slight difference for different RNA sequencing experiments, the protocol is generally similar and involves these steps:

l The complete set of transcripts of the target cell or tissue should be isolated from the sample.

l If a specific type of RNA is to be profiled, researchers need to purify and enrich that kind of RNA from the total RNA molecules.

l Make an RNA sequencing library that consists of a pool of complementary DNA (cDNA) fragments with adapters attached.

l Perform sequence with the next-generation sequencing platform.

l Finally analyze the short-read sequencing result.

Before being sequenced by the next-generation sequencing platform, the RNA sample is required to be fragmented, end-repaired, and collected into adapter-ligated libraries. The RNA-Seq library preparation is dependent on the NGS platform applied and can influence the results generated by NGS experiments. Fortunately, most service providers will offer library preparation kits for directional and non-directional RNA library prep that match their platforms for sequencing with stable reagents and streamlined workflows.

Preparation of RNA Sequencing Library for NGS

There are two common library preparation methods, ligation-based library prep and tagmentation-based library prep. The ligation method separates the steps of DNA fragmentation and ligation of adapters to the ends of the fragments, while the tagmentation method combines DNA fragmentation and adapter ligation into one reaction step. Take ligation-based library preparation as an example, here are the two major steps required to prepare an RNA sequencing library.

l Addition of adapters

A ligase enzyme covalently will link the adapter and insert cDNA fragments, making a complete library molecule. These adapters have different functions so their sequences specific to the NGS platform should also be attached to the flow cell to allow RNA sequencing.

l PCR amplification

Amplification of libraries is optional and depends on the adapter type and input used. The remaining oligonucleotides and small fragments should be removed using magnetic beads or a spin column if PCR amplification is applied.

Besides, researchers might need to consider reverse transcription of RNA and cDNA synthesis and whether or not to preserve strandedness. The adaptor sequences that will be specific to the sequencing platform must also be added to the end of the fragments. Once the RNA sequencing library gets prepared, researchers can go ahead with the next step in the next-generation sequencing workflow.

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(via Exosome Engineering Services - Creative Biolabs)

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Creative Biolabs provides comprehensive exosome-related services to global clients.

With years of experience in the field of molecular and cell biology research, Creative Biolabs is capable of supporting all aspects of your exosome research. Equipped with state-of-the-art tools and technologies, we offer the most comprehensive exosome-related research services, ranging from isolation and characterization to cargo profiling and engineering, for a better understanding of their physiological functions, for the discovery of clinical biomarkers, and engineering of exosomes for therapeutic applications.