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Bromelain

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Laura P Hale – 1st expert on this subject based on the ideXlab platform

  • Bromelain treatment decreases neutrophil migration to sites of inflammation
    Clinical Immunology, 2008
    Co-Authors: David J Fitzhugh, Siqing Shan, Mark W Dewhirst, Laura P Hale

    Abstract:

    Abstract Bromelain, a mixture of proteases derived from pineapple stem, has been reported to have therapeutic benefits in a variety of inflammatory diseases, including murine inflammatory bowel disease. The purpose of this work was to understand potential mechanisms for this anti-inflammatory activity. Exposure to Bromelain in vitro has been shown to remove a number of cell surface molecules that are vital to leukocyte trafficking, including CD128a/CXCR1 and CD128b/CXCR2 that serve as receptors for the neutrophil chemoattractant IL-8 and its murine homologues. We hypothesized that specific proteolytic removal of CD128 molecules by Bromelain would inhibit neutrophil migration to IL-8 and thus decrease acute responses to inflammatory stimuli. Using an in vitro chemotaxis assay, we demonstrated a 40% reduction in migration of Bromelain– vs. sham-treated human neutrophils in response to rhIL-8. Migration to the bacterial peptide analog fMLP was unaffected, indicating that Bromelain does not induce a global defect in leukocyte migration. In vivo Bromelain treatment generated a 50–85% reduction in neutrophil migration in 3 different murine models of leukocyte migration into the inflamed peritoneal cavity. Intravital microscopy demonstrated that although in vivo Bromelain treatment transiently decreased leukocyte rolling, its primary long-term effect was abrogation of firm adhesion of leukocytes to blood vessels at the site of inflammation. These changes in adhesion were correlated with rapid re-expression of the Bromelain-sensitive CD62L/L-selectin molecules that mediate rolling following in vivo Bromelain treatment and minimal re-expression of CD128 over the time period studied. Taken together, these studies demonstrate that Bromelain can effectively decrease neutrophil migration to sites of acute inflammation and support the specific removal of the CD128 chemokine receptor as a potential mechanism of action.

  • Bromelain treatment decreases secretion of pro inflammatory cytokines and chemokines by colon biopsies in vitro
    Clinical Immunology, 2008
    Co-Authors: Jane E Onken, Paula K Greer, Brian Calingaert, Laura P Hale

    Abstract:

    Abstract Oral Bromelain has been anecdotally reported to decrease inflammation in ulcerative colitis (UC). Proteolytically active Bromelain is known to decrease expression of mRNAs encoding pro-inflammatory cytokines by human leukocytes in vitro. To assess the effect of Bromelain on mucosal secretion of cytokines in inflammatory bowel disease (IBD), endoscopic colon biopsies from patients with UC, Crohn’s disease (CD), and non-IBD controls were treated in vitro with Bromelain or media, then cultured. Secretion of pro-inflammatory cytokines and chemokines was measured. Significant increases in granulocyte colony-stimulating factor (G-CSF), interferon (IFN)-γ, interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF) were detected in the media from actively inflamed areas in UC and CD as compared with non-inflamed IBD tissue and non-IBD controls. In vitro Bromelain treatment decreased secretion of G-CSF, granulocyte–macrophage colony-stimulating factor (GM-CSF), IFN-γ, CCL4/macrophage inhibitory protein (MIP)-1β, and TNF by inflamed tissue in IBD. Bromelain may be a novel therapy for IBD.

  • Oral immunogenicity of the plant proteinase Bromelain.
    International immunopharmacology, 2006
    Co-Authors: Laura P Hale, David J Fitzhugh, Herman F Staats

    Abstract:

    Bromelain is a natural mixture of proteolytic enzymes derived from pineapple stem that has been shown to have anti-inflammatory activity when administered orally. Although most proteins given orally without adjuvant (e.g., food) result in tolerance, we previously reported that long-term oral exposure to Bromelain stimulated the development of high serum anti-Bromelain antibody titers. The purpose of these studies was to further investigate the mechanisms responsible for the immunogenicity of oral Bromelain. Results showed that repeated exposure was required for development of anti-Bromelain antibodies, with strong antibody responses in all mice that received at least 12 doses of Bromelain either orally or intragastrically over 3-6 weeks. Proteolytic activity was required for strong oral immunogenicity in the absence of conventional adjuvant, with strong serum antibody responses generated against proteolytically active Bromelain and trypsin, but not against ovalbumin, lysozyme, or inactivated Bromelain. Significantly higher anti-Bromelain antibody titers were seen in IL-10-deficient versus wild-type mice, suggesting that simultaneous treatments that decrease IL-10 activity may further enhance systemic antibody responses following oral exposure. The antibodies generated did not affect the proteolytic activity of Bromelain. The data demonstrate that proteolytically active antigens such as Bromelain can stimulate both systemic and mucosal immune responses following repeated oral exposure. Further studies of the mechanisms involved in generation of immune responses following oral exposure to proteolytically active antigens can lead to a better understanding of mechanisms of oral tolerance and to the development of novel adjuvants for oral vaccines.

Yogeshwer Shukla – 2nd expert on this subject based on the ideXlab platform

  • Bromelain inhibits nuclear factor kappa b translocation driving human epidermoid carcinoma a431 and melanoma a375 cells through g2 m arrest to apoptosis
    Molecular Carcinogenesis, 2012
    Co-Authors: Kulpreet Bhui, Shilpa Tyagi, Amit Srivastava, Madhulika Singh, Richa Singh, Yogeshwer Shukla

    Abstract:

    Bromelain, obtained from pineapple, is already in use clinically as adjunct in chemotherapy. Our objective was to test its ability to act as a sole anti-cancer agent. Therefore, we describe its anti-proliferative, anti-inflammatory and subsequent anti-cancer effects in vitro, against human epidermoid carcinoma-A431 and melanoma-A375 cells. Bromelain exhibited reduction in proliferation of both these cell-lines and suppressed their potential for anchorage-independent growth. Further, suppression of inflammatory signaling by Bromelain was evident by inhibition of Akt regulated-nuclear factor-kappaB activation via suppression of inhibitory-kappaBα phosphorylation and concomitant reduction in cyclooxygenase-2. Since, the inflammatory cascade is well-known to be closely allied to cancer; we studied the effect of Bromelain on events/molecules central to it. Bromelain caused depletion of intracellular glutathione and generation of reactive oxygen-species followed by mitochondrial membrane depolarization. This led to Bromelain-induced cell-cycle arrest at G2/M phase which was mediated by modulation of cyclin B1, phospho-cdc25C, Plk1, phospho-cdc2, and myt1. This was subsequently followed by induction of apoptosis, indicated by membrane-blebbing, modulation of Bax-Bcl-2 ratio, Apaf-1, caspase-9, and caspase-3; chromatin-condensation, increase in caspase-activity and DNA-fragmentation. Bromelain afforded substantial anti-cancer potential in these settings; hence we suggest it as a potential prospect for anti-cancer agent besides only an additive in chemotherapy. © 2011 Wiley Periodicals, Inc.

  • pineapple Bromelain induces autophagy facilitating apoptotic response in mammary carcinoma cells
    Biofactors, 2010
    Co-Authors: Kulpreet Bhui, Shilpa Tyagi, Bharti Prakash, Yogeshwer Shukla

    Abstract:

    Bromelain, from pineapple, possesses potent anticancer effects. We investigated autophagic phenomenon in mammary carcinoma cells (estrogen receptor positive and negative) under Bromelain treatment and also illustrated the relationship between autophagy and apoptosis in MCF- 7 cells. MCF- 7 cells exposed to Bromelain showed delayed growth inhibitory response and induction of autophagy, identified by monodansylcadaverine localization. It was succeeded by apoptotic cell death, evident by sub-G1 cell fraction and apoptotic features like chromatin condensation and nuclear cleavage. 3 -Methyladenine (MA, autophagy inhibitor) pretreatment reduced the Bromelain-induced autophagic level, also leading to decline in apoptotic population, indicating that here autophagy facilitates apoptosis. However, addition of caspase- 9 inhibitor Z-LEHD-FMK augmented the autophagy levels, inhibited morphological apoptosis but did not prevent cell death. Next, we found that Bromelain downregulated the phosphorylation of extracellular signal-regulated kinase ½ (ERK½), whereas that of c-jun N-terminal kinase (JNK) and p38 kinase were upregulated. Also, MA had no influence on Bromelain-suppressed ERK½ activation, yet, it downregulated JNK and p38 activation. Also, addition of mitogen-activated protein kinase (MAPK) inhibitors enhanced the autophagic ratios, which suggested the role of MAP kinases in Bromelain-induced autophagy. All three MAPKs were seen to be constantly activated over the time. Bromelain was seen to induce the expressions of autophagy-related proteins, light chain 3 protein B II (LC 3 BII), and beclin-1. Using ERK½ inhibitor, expressions of LC 3 BII and beclin-1 increased, whereas p38 and JNK inhibitors decreased this protein expression, indicating that Bromelain-induced autophagy was positively regulated by p38 and JNK but negatively regulated by ERK½. Autophagy-inducing property of Bromelain can be further exploited in breast cancer therapy.

  • Bromelain inhibits cox 2 expression by blocking the activation of mapk regulated nf kappa b against skin tumor initiation triggering mitochondrial death pathway
    Cancer Letters, 2009
    Co-Authors: Kulpreet Bhui, Sahdeo Prasad, Jasmine George, Yogeshwer Shukla

    Abstract:

    Abstract Chemoprevention impels the pursuit for either single targeted or cocktail of multi-targeted agents. Bromelain, potential agent in this regard, is a pharmacologically active compound, present in stems and fruits of pineapple ( Ananas cosmosus ), endowed with anti-inflammatory, anti-invasive and anti-metastatic properties. Herein, we report the anti tumor-initiating effects of Bromelain in 2-stage mouse skin tumorigenesis model. Pre-treatment of Bromelain resulted in reduction in cumulative number of tumors (CNT) and average number of tumors per mouse. Preventive effect was also comprehended in terms of reduction in tumor volume up to a tune of ∼65%. Components of the cell signaling pathways, connecting proteins involved in cell death were targeted. Bromelain treatment resulted in upregulation of p53 and Bax and subsequent activation of caspase 3 and caspase 9 with concomitant decrease in Bcl-2. A marked inhibition in cyclooxygenase-2 (Cox-2) expression and inactivation of nuclear factor-kappa B (NF-κB) was recorded, as phosphorylation and consequent degradation of Iκ Bα was blocked by Bromelain. Also, Bromelain treatment curtailed extracellular signal regulated protein kinase (ERK1/2), p38 mitogen-activated protein kinase (MAPK) and Akt activity. The basis of anti tumor-initiating activity of Bromelain was revealed by its time dependent reduction in DNA nick formation and increase in percentage prevention. Thus, modulation of inappropriate cell signaling cascades driven by Bromelain is a coherent approach in achieving chemoprevention.

Azura Amid – 3rd expert on this subject based on the ideXlab platform

  • Functional characterization of recombinant Bromelain
    , 2015
    Co-Authors: Azura Amid, Muntari Bala, Hamzah Mohd. Salleh

    Abstract:

    Stem Bromelain is a plant protease with a number of industrial and therapeutic
    applications. This study investigated the functional properties of commercial Bromelain
    and recombinant Bromelain (expressed in E. coli BL 21-AI). In vitro cytotoxicity assays
    were conducted to evaluate the effect of both Bromelains on tumour cell lines (murine
    melanoma and breast cancer MCF-7) and normal cells (Vero and Chinese hamster
    ovary) using Sulforhodamine B and MTT tests. Anti-inflammatory activities of the
    enzymes were also assessed on murine macrophage cell lines using Griess reagents and
    ELISA test. The beef tenderizing effects of both Bromelains and in-situ localization of the
    recombinant Bromelain were analysed using Transmission Electron Microscopy. The
    results obtained revealed that both Bromelains were more cytotoxic to tumour cell lines
    (IC50 values ≈ 0.16-0.23mg/mL) than normal cell lines (IC50 values ≈ 0.27-0.51mg/mL).
    Moreover, the two enzymes had effectively inhibited the pro-inflammatory mediators’
    production and thus, had good anti-inflammatory activity. Both enzymes were found to
    have comparative anti-cancer and anti-inflammatory properties. The expression of
    recombinant enzyme was discovered to occur in the cytoplasm of E. coli BL 21-AI and
    the purified enzyme efficiently degraded both actin and myosin of fresh beef. Hence, it
    served as a good meat tenderizer.

  • Case Study: Recombinant Bromelain Downstream Processing
    Recombinant Enzymes – From Basic Science to Commercialization, 2015
    Co-Authors: Azura Amid, Zatul Iffaf Mohd Arshad, Muhd. Ezza Faiez Othman

    Abstract:

    This chapter presents details on the purification, formulation and drying of recombinant Bromelain. The wide range of applications of recombinant Bromelain has increased interest in finding viable purification techniques for large-scale production. An affinity chromatography technique was developed by Amid and co-workers (Expression, purification, and characterization of a recombinant stem Bromelain from Ananas comosus, Process Biochem 46:2232–2239, 2011) to purify recombinant Bromelain. However, this technique presented low recovery and small sample loading capacity and thus is not suitable as a purification tool in the large-scale production of recombinant Bromelain. An aqueous two-phase system is one alternative method that we use to purify recombinant Bromelain, as it reliable and easy to scale up and has a low cost. As part of avoiding cysteine degradation, spray drying the purified recombinant protein with maltodextrin as an excipient provides the possibility of preserving its activity and creating fine particles that are suitable for end-product application. The processes of the purification, formulation and spray drying of recombinant Bromelain are explained briefly.

  • Bromelain: an overview of industrial application and purification strategies
    Applied Microbiology and Biotechnology, 2014
    Co-Authors: Zatul Iffah Mohd Arshad, Azura Amid, Faridah Yusof, Irwandi Jaswir, Kausar Ahmad, Show Pau Loke

    Abstract:

    This review highlights the use of Bromelain in various applications with up-to-date literature on the purification of Bromelain from pineapple fruit and waste such as peel, core, crown, and leaves. Bromelain, a cysteine protease, has been exploited commercially in many applications in the food, beverage, tenderization, cosmetic, pharmaceutical, and textile industries. Researchers worldwide have been directing their interest to purification strategies by applying conventional and modern approaches, such as manipulating the pH, affinity, hydrophobicity, and temperature conditions in accord with the unique properties of Bromelain. The amount of downstream processing will depend on its intended application in industries. The breakthrough of recombinant DNA technology has facilitated the large-scale production and purification of recombinant Bromelain for novel applications in the future.