scholarly journals Αλληλεπίδραση μεταξύ φυσικής και ειδικής ανοσίας στο επίπεδο των Toll-like και TCR υποδοχέων, σε λοιμώξεις από παράσιτα του γένους Leishmania

2015 ◽  
Author(s):  
Όλγα Κουτσώνη
Keyword(s):  
In Silico ◽  
Reverse Vaccinology ◽  
Tnf Α ◽  
Leishmania Spp

Οι Th1 ανοσολογικές αποκρίσεις συμβάλλουν στην ανάπτυξη προστατευτικής ανοσίας έναντι της λεϊσμανίασης. Τα τελευταία χρόνια, έχουν προσδιοριστεί διάφορα ανασυνδυασμένα αντιγόνα του παρασίτου Leishmania spp., τα οποία επέδειξαν ικανότητα επαγωγής Th1 ανοσολογικής απόκρισης και συνεπώς, υποσχόμενη δυναμική για την κατασκευή αποτελεσματικού εμβολίου έναντι της λεϊσμανίασης. Οι ευκαρυωτικοί παράγοντες έναρξης eIF του παρασίτου Leishmania spp. (LeIF), αποτελούν υψηλά συντηρημένες πρωτεΐνες και μελέτες αναφέρουν πως οι ευκαρυωτικοί παράγοντες έναρξης των στελεχών L. braziliensis και L. major, επάγουν αποτελεσματική προστασία έναντι της λεϊσμανίασης, σε πειραματικά μοντέλα ποντικών. Στην παρούσα διδακτορική διατριβή, διερευνήθηκε η αντιλεϊσμανιακή δράση του ευκαρυωτικού παράγοντα έναρξης του στελέχους L. infantum (LieIF) και αποδείχθηκε για πρώτη φορά η προστατευτική ή/και ανοσοθεραπευτική δράση της πρωτεΐνης, σε in vitro σύστημα L. donovani-μολυσμένων μακροφάγων. Επίσης, προσδιορίστηκαν οι δραστικοί μηχανισμοί που επιστρατεύουν τα μακροφάγα ώστε να καταστείλουν τον πολλαπλασιασμό του παρασίτου και επιβεβαιώθηκε πως η λεϊσμανιοκτόνος δράση της LieIF, διαμεσολαβείται από την παραγωγή των παραγόντων NO και ROS. Αυτή η προστατευτική δράση της LieIF συσχετίστηκε με την ενεργοποίηση ενός TNF-α εξαρτώμενου μονοπατιού καθώς και με την παραγωγή της κυτταροκίνης IL-12. Αντιθέτως, η αντίστοιχη παραγωγή των αντιμικροβιακών μορίων, που προκλήθηκε από την ανοσοθεραπευτική δράση της LieIF, αποδείχθηκε πως δεν ρυθμίζεται από μηχανισμούς που περιλαμβάνουν τη συμμετοχή του TNF-α. Στη συνέχεια, μελετήθηκαν οι ανοσοτροποποιητικές ιδιότητες της LieIF, σε in vitro σύστημα μυελοειδών δενδριτικών κυττάρων (ΔΚ), τόσο σε επίπεδο ωρίμανσης και λειτουργικής διαφοροποίησης των κυττάρων, όσο και σε επίπεδο συνέργειας με τους υποδοχείς τύπου Toll (TLRs). Η επιβεβαίωση της ενίσχυσης των ανοσοτροποποιητικών ιδιοτήτων της ανασυνδυασμένης LieIF από τη συνεργιστική δράση των TLRs, ενισχύει τη μελλοντική χρήση της πρωτεΐνης σε πειραματικά πρωτόκολλα ανοσοθεραπείας με ενισχυμένο θεραπευτικό αποτέλεσμα. Επιπλέον, ακολουθώντας την μεθοδολογία της αντίστροφης πορείας σχεδιασμού εμβολίων (reverse vaccinology) πραγματοποιήθηκε in silico ανάλυση της LieIF, προκειμένου να προσδιοριστούν τα ελάχιστα δυνατά τμήματά της, που αναγνωρίζονται εν δυνάμει από τους ομόλογους υποδοχείς των Τ κυττάρων (TCR) και φέρουν ανοσογονικές ιδιότητες. Η in silico ανάλυση των πρωτεϊνών με τη χρήση αλγορίθμων πρόβλεψης αντιγονικότητας επιταχύνει τη διαδικασία εύρεσης αντιγονικών επιτόπων, ενισχύοντας την έρευνα που σχετίζεται με τον προσδιορισμό κατάλληλων πεπτιδίων για πρότυπα πρωτόκολλα συνθετικών εμβολίων. Τα συνθετικά πεπτίδια που προέκυψαν από την παραπάνω διαδικασία, καθώς και μεγαλύτερα αλληλοεπικαλυπτόμενα ανασυνδυασμένα πρωτεΐνικά τμήματα της LieIF, ελέγχθηκαν περαιτέρω, ως προς τις ανοσοτροποποιητικές τους ιδιότητες, σε in vitro συστήματα μακροφάγων και ΔΚ. Τα αποτελέσματα έδειξαν για πρώτη φορά, την ύπαρξη πολυεπιτοπικών τμημάτων της πρωτεΐνης με ισχυρές ανοσοτροποποιητικές ιδιότητες. Τα in vitro αποτελέσματα οδήγησαν στην αξιολόγηση της in vivo δράσης της LieIF σε πειραματικά μοντέλα ευαίσθητων έναντι της λεϊσμανίασης, ποντικών. Ειδικότερα, μελετήθηκε η δράση της LieIF ως ανοσοενισχυτικό και αναπτύχθηκαν πειραματικά πρωτόκολλα ανοσοθεραπείας με διαλυτή πρωτεΐνη και πρωτόκολλα προστατευτικού εμβολιασμού με ανοσιακή μεταφορά δενδριτικών κυττάρων ως «οχήματα μεταφοράς» της LieIF. Συνοψίζοντας, η LieIF επέδειξε ιδιότητες και δραστικότητα ανοσοενισχυτικού, ικανού να προάγει προσαρμοστική ανοσία, καθώς και ισχυρές ανοσοτροποποιητικές ιδιότητες σε in vitro και in vivo συστήματα, που αποδείχθηκαν ικανές να προσανατολίσουν την πόλωση μιας Th1-διαμεσολαβούμενης ανοσολογικής απόκρισης. Τα ευρήματα της μελέτης αυτής προσφέρουν πληρέστερη κατανόηση του ρόλου αυτής της κυτταροπλασματικής πρωτεΐνης του παρασίτου, αναδεικνύοντας τη μελλοντική της εκμετάλλευση στη στρατηγική ενός αποτελεσματικού ελέγχου της νόσου.

scholarly journals Anti-Arthritic and Anti-Inflammatory Potential of Spondias mangifera Extract Fractions: An In Silico, In Vitro and In Vivo Approach

Plants ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 825
Author(s):  
Mohammad Khalid ◽  
Mohammed H. Alqarni ◽  
Ambreen Shoaib ◽  
Muhammad Arif ◽  
Ahmed I. Foudah ◽  
...  
Keyword(s):  
Molecular Docking ◽  
In Silico ◽  
Arthritis Score ◽  
Beneficial Effects ◽  
Tnf Α ◽  
Cox 2 ◽  
Anti Inflammatory ◽  
Cox 1

The fruits of Spondias mangifera (S. mangifera) have traditionally been used for the management of rheumatism in the northeast region of India. The present study explores the probable anti-arthritis and anti-inflammatory potential of S. mangifera fruit extract’s ethanolic fraction (EtoH-F). To support this study, we first approached the parameters in silico by means of the active constituents of the plant (beta amyrin, beta sitosterol, oleonolic acid and co-crystallised ligands, i.e., SPD-304) via molecular docking on COX-1, COX-2 and TNF-α. Thereafter, the absorption, distribution, metabolism, excretion and toxicity properties were also determined, and finally experimental activity was performed in vitro and in vivo. The in vitro activities of the plant extract fractions were evaluated by means of parameters like 1,1-Diphenyl-2- picrylhydrazyl (DPPH), free radical-reducing potential, albumin denaturation, and protease inhibitory activity. The in vivo activity was evaluated using parameters like COX, TNF-α and IL-6 inhibition assay and arthritis score in Freund Adjuvant (CFA) models at a dose of 400 mg/kg b.w. per day of different fractions (hexane, chloroform, alcoholic). The molecular docking assay was performed on COX-1, COX-2 and TNF-α. The results of in vitro studies showed concentration-dependent reduction in albumin denaturation, protease inhibitors and scavenging activity at 500 µg/mL. Administration of the S. mangifera alcoholic fraction at the abovementioned dose resulted in a significant reduction (p < 0.01) in arthritis score, paw diameters, TNF-α, IL-6 as compared to diseased animals. The docking results showed that residues show a critical binding affinity with TNF-α and act as the TNF-α antagonist. The alcoholic fraction of S. mangifera extract possesses beneficial effects on rheumatoid arthritis as well as anti-inflammatory potential, and can further can be used as a possible agent for novel target-based therapies for the management of arthritis.

Activity of a hydroxybibenzyl bryophyte constituent against Leishmania spp. and Trypanosoma cruzi: In silico, in vitro and in vivo activity studies

2008 ◽  
Vol 43 (9) ◽  
pp. 1797-1807 ◽  
Author(s):  
Virginia Roldos ◽  
Hector Nakayama ◽  
Miriam Rolón ◽  
Alina Montero-Torres ◽  
Fernando Trucco ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
In Silico ◽  
Leishmania Spp

scholarly journals Validating potent anti-inflammatory and anti-rheumatoid properties of Drynaria quercifolia rhizome methanolic extract through in vitro, in vivo, in silico and GC-MS-based profiling

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Debabrata Modak ◽  
Subhashis Paul ◽  
Sourav Sarkar ◽  
Subarna Thakur ◽  
Soumen Bhattacharjee
Keyword(s):  
Rheumatoid Arthritis ◽  
In Silico ◽  
Methanolic Extract ◽  
Paw Edema ◽  
Active Components ◽  
Tnf Α ◽  
Cox 2 ◽  
Anti Inflammatory

Abstract Background The fronds of Drynaria quercifolia have traditionally been used in rheumatic pain management. The goal of the present study was to validate the potent anti-inflammatory and anti-rheumatoid properties of the methanolic-extract of its rhizome using in vitro, in vivo and in silico strategies. Methods The plant was collected and the methanolic extract was prepared from its rhizome. Protein denaturation test, hypotonicity and heat-induced haemolysis assays were performed in vitro. The in vivo anti-rheumatoid potential was assessed in Freund’s complete adjuvant (FCA)-induced Wistar rat model through inflammatory paw-edema, haematological, biochemical, radiological and histopathological measurements. Moreover, metabolites of methanolic extract were screened by gas chromatography-mass spectrometry (GC-MS) and 3D molecular structures of active components were utilized for in silico docking study using AutoDock. Results In vitro results evinced a significant (p < 0.05) anti-inflammatory activity of the rhizome methanolic extract in a dose-linear response. Further, Drynaria quercifolia rhizome methanolic extract (DME) significantly ameliorated rheumatoid arthritis as indicated by the inhibition of arthritic paw-edema (in millimeter) in the rat rheumatoid arthritis models in both the low (57.71 ± 0.99, p < 0.01) and high dose groups (54.45 ± 1.30, p < 0.001) when compared to arthritic control. Treatment with DME also normalized the haematological (RBC, WBC, platelet counts and hemoglobin contents) and biochemical parameters (total protein, albumin, creatinine and ceruloplasmin) significantly (p < 0.05), which were further supported by histopathological and radiological analyses. Furthermore, GC-MS analysis of DME demonstrated the presence of 47 phytochemical compounds. Compounds like Squalene, Gamma Tocopherol, n-Hexadecanoic acid showed potent inhibition of cyclooxygenase-2 (COX-2), tumor necrosis factor (TNF-α), and interleukin (IL-6) in the docking analysis. Conclusion Results from in vivo and in vitro studies indicated that DME possesses a potent anti-inflammatory and anti-arthritic activity. In silico studies delineated the emergent potent inhibitory effects of several bio-active components on the target inflammatory markers (COX-2, TNF-α and IL-6).

Trigonella foenum-graecum Seeds Oil Attenuated Inflammation and Angiogenesis In Vivo through Down-Regulation of TNF-α

2020 ◽  
Vol 20 ◽  
Author(s):  
Muhammad Asif ◽  
Hafiz Muhammad Yousaf ◽  
Mohammed Saleem ◽  
Malik Saadullah ◽  
Tahir Ali Chohan ◽  
...  
Keyword(s):  
In Silico ◽  
Docking Studies ◽  
Antioxidant Potential ◽  
Cam Assay ◽  
In Silico Docking ◽  
Trigonella Foenum Graecum ◽  
Tnf Α ◽  
Anti Inflammatory

Introduction: Inflammation is a vital reaction of the natural immune system that protects against encroaching agents. However, uncontrolled inflammation can lead to complications. Trigonella foenum-graecum is traditionally used as an anti-inflammatory herb. Objectives: The current study was conducted to explore the antioxidant, anti-inflammatory, and antiangiogenic potentials of Trigonella foenum-graecum seeds oil. Methods: Oil was extracted from seeds of Trigonella foenum-graecum by cold press method and labelled as TgSO. Phytochemical (GCMS, Folin-Ciocalteu method) and metal analyses were conducted to evaluate the metalo-chemical profile of TgSO. In vitro antioxidant assays (2,2-diphenyl-1-picrylhydrazyl, 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid and ferric reducing antioxidant power) were performed to assess its antioxidant potential. In vitro antimicrobial property was evaluated using the agar disc diffusion method and the safety profile of TgSO was assessed following OECD 425 guidelines. In vivo anti-inflammatory activity of TgSO was assessed in carrageenan, serotonin, histamine, formalin, and cotton pellet-induced oedema models. Serum TNF-α, superoxide dismutase (SOD) and, catalases (CAT) levels were assessed by ELISA kit while the effects on angiogenesis were assessed by chick chorioallantoic membrane (CAM) assay. Histopathological studies using excised paws were conducted to observe the effect of TgSO treatment at the tissue level. In silico docking studies were conducted to screen binding potential of identified compounds towards TNF- α. Results: Extraction by cold press yielded 16% of TgSO. Phytochemical analysis of TgSO through GC-MS showed the presence of eugenol, dihydrocoumairn, and heptadecanoic acid, tri- and tetradecanoic acid and hexadecanoic acid respectively. Total phenolic contents of TgSO were found to be 37.1 ± 0.91 mg/g gallic acid equivalent in Folin-Ciocalteu method. Metal analysis indicated the presence of different metals in TgSO. Findings of antioxidant models showed moderate antioxidant potential of TgSO. Findings of antimicrobial assays showed that TgSO was active against S. aureus, S.epidermidis, C. albicans, and A. niger. In vivo toxicity study data showed that TgSO was safe up to the dose of 5000 mg/kg. Data of oedema models showed significant (p < 0.05) reduction in oedema development in TgSO treated animals in both acute and chronic models. Histopathological evaluations of paws showed minimal infiltration with inflammatory cells in TgSO-treated animals. Treatment also significantly (p < 0.05) down-regulated TNF-α in serum and while levels of SOD and CAT were upregulated. CAM assay findings revealed antiangiogenic activity of TgSO. Findings of in silico docking studies showed that identified phytoconstituents have potential to bind with culprit cytokine. Conclusion: Data of that current study conclude that TgSO has antioxidant, anti-inflammatory, and antiangiogenic effects that validate its traditional uses. Moreover, the synergistic actions of different phytoconstituents are proposed to be responsible for the observed effects.

Recent In Vitro and In Silico Advances in the Understanding of Intranasal Drug Delivery

2020 ◽  
Vol 26 ◽  
Author(s):  
John Chen ◽  
Andrew Martin ◽  
Warren H. Finlay
Keyword(s):  
Drug Delivery ◽  
In Silico ◽  
Local Drug Delivery ◽  
In Vivo Studies ◽  
Intranasal Drug Delivery ◽  
Nasal Sprays ◽  
In Silico Studies ◽  
Drug Delivery Devices

Background: Many drugs are delivered intranasally for local or systemic effect, typically in the form of droplets or aerosols. Because of the high cost of in vivo studies, drug developers and researchers often turn to in vitro or in silico testing when first evaluating the behavior and properties of intranasal drug delivery devices and formulations. Recent advances in manufacturing and computer technologies have allowed for increasingly realistic and sophisticated in vitro and in silico reconstructions of the human nasal airways. Objective: To perform a summary of advances in understanding of intranasal drug delivery based on recent in vitro and in silico studies. Conclusion: The turbinates are a common target for local drug delivery applications, and while nasal sprays are able to reach this region, there is currently no broad consensus across the in vitro and in silico literature concerning optimal parameters for device design, formulation properties and patient technique which would maximize turbinate deposition. Nebulizers are able to more easily target the turbinates, but come with the disadvantage of significant lung deposition. Targeting of the olfactory region of the nasal cavity has been explored for potential treatment of central nervous system conditions. Conventional intranasal devices, such as nasal sprays and nebulizers, deliver very little dose to the olfactory region. Recent progress in our understanding of intranasal delivery will be useful in the development of the next generation of intranasal drug delivery devices.

Naphthoquinones and derivatives for chemotherapy: perspectives and limitations of their anti-trypanosomatids activities

2020 ◽  
Vol 26 ◽  
Author(s):  
Luíza Dantas-Pereira ◽  
Edézio F. Cunha-Junior ◽  
Valter V. Andrade-Neto ◽  
John F. Bower ◽  
Guilherme A. M. Jardim ◽  
...  
Keyword(s):  
Large Scale ◽  
Neglected Tropical Diseases ◽  
Folk Medicine ◽  
Leishmania Species ◽  
Parasitic Infections ◽  
Mechanistic Analysis ◽  
Leishmania Spp ◽  
Nanomolar Range

: Chagas disease, Sleeping sickness and Leishmaniasis, caused by trypanosomatids Trypanosoma cruzi, Trypanosoma brucei and Leishmania spp., respectively, are considered neglected tropical diseases, and they especially affect impoverished populations in the developing world. The available chemotherapies are very limited and a search for alternatives is still necessary. In folk medicine, natural naphthoquinones have been employed for the treatment of a great variety of illnesses, including parasitic infections. This review is focused on the anti-trypanosomatid activity and mechanistic analysis of naphthoquinones and derivatives. Among all the series of derivatives tested in vitro, naphthoquinone-derived 1,2,3-triazoles were very active on T. cruzi infective forms in blood bank conditions, as well as in amastigotes of Leishmania spp. naphthoquinones containing a CF3 on a phenyl amine ring inhibited T. brucei proliferation in the nanomolar range, and naphthopterocarpanquinones stood out for their activity on a range of Leishmania species. Some of these compounds showed a promising selectivity index (SI) (30 to 1900), supporting further analysis in animal models. Indeed, high toxicity to the host and inactivation by blood components are crucial obstacles to be overcome to use naphthoquinones and/or their derivatives for chemotherapy. Multidisciplinary initiatives embracing medicinal chemistry, bioinformatics, biochemistry, and molecular and cellular biology need to be encouraged to allow the optimization of these compounds. Large scale automated tests are pivotal for the efficiency of the screening step, and subsequent evaluation of both the mechanism of action in vitro and pharmacokinetics in vivo are essential for the development of a novel, specific and safe derivative, minimizing adverse effects.

In-silico Studies of Isolated Phytoalkaloid Against Lipoxygenase: Study Based on Possible Correlation

2018 ◽  
Vol 21 (3) ◽  
pp. 215-221
Author(s):  
Haroon Khan ◽  
Muhammad Zafar ◽  
Helena Den-Haan ◽  
Horacio Perez-Sanchez ◽  
Mohammad Amjad Kamal
Keyword(s):  
In Silico ◽  
Docking Studies ◽  
In Vivo Studies ◽  
In Vitro Assays ◽  
Chronic Obstructive ◽  
Lipoxygenase Inhibitors ◽  
Lipoxygenase Inhibition ◽  
In Silico Studies

Aim and Objective: Lipoxygenase (LOX) enzymes play an important role in the pathophysiology of several inflammatory and allergic diseases including bronchial asthma, allergic rhinitis, atopic dermatitis, allergic conjunctivitis, rheumatoid arthritis and chronic obstructive pulmonary disease. Inhibitors of the LOX are believed to be an ideal approach in the treatment of diseases caused by its over-expression. In this regard, several synthetic and natural agents are under investigation worldwide. Alkaloids are the most thoroughly investigated class of natural compounds with outstanding past in clinically useful drugs. In this article, we have discussed various alkaloids of plant origin that have already shown lipoxygenase inhibition in-vitro with possible correlation in in silico studies. Materials and Methods: Molecular docking studies were performed using MOE (Molecular Operating Environment) software. Among the ten reported LOX alkaloids inhibitors, derived from plant, compounds 4, 2, 3 and 1 showed excellent docking scores and receptor sensitivity. Result and Conclusion: These compounds already exhibited in vitro lipoxygenase inhibition and the MOE results strongly correlated with the experimental results. On the basis of these in vitro assays and computer aided results, we suggest that these compounds need further detail in vivo studies and clinical trial for the discovery of new more effective and safe lipoxygenase inhibitors. In conclusion, these results might be useful in the design of new and potential lipoxygenase (LOX) inhibitors.

In Silico Assessment of ADME Properties: Advances in Caco-2 Cell Monolayer Permeability Modeling

2019 ◽  
Vol 18 (26) ◽  
pp. 2209-2229 ◽  
Author(s):  
Hai Pham-The ◽  
Miguel Á. Cabrera-Pérez ◽  
Nguyen-Hai Nam ◽  
Juan A. Castillo-Garit ◽  
Bakhtiyor Rasulev ◽  
...  
Keyword(s):  
Intestinal Absorption ◽  
In Silico ◽  
Review Paper ◽  
Cell Monolayer ◽  
Cell Permeability ◽  
Drug Substances ◽  
Wide Range ◽  
Modeling Techniques

One of the main goals of in silico Caco-2 cell permeability models is to identify those drug substances with high intestinal absorption in human (HIA). For more than a decade, several in silico Caco-2 models have been made, applying a wide range of modeling techniques; nevertheless, their capacity for intestinal absorption extrapolation is still doubtful. There are three main problems related to the modest capacity of obtained models, including the existence of inter- and/or intra-laboratory variability of recollected data, the influence of the metabolism mechanism, and the inconsistent in vitro-in vivo correlation (IVIVC) of Caco-2 cell permeability. This review paper intends to sum up the recent advances and limitations of current modeling approaches, and revealed some possible solutions to improve the applicability of in silico Caco-2 permeability models for absorption property profiling, taking into account the above-mentioned issues.

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