The acute effects of nonstructural-1 protein dengue virus type 2 on wet liver weight, zonulin expression and serum zonulin
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Abstract
Background and Objectives: Intestinal leakage commonly occurs in severe dengue infection with zonulin as a biomarker. The aim of this study was to determine the effects of NS1 on liver weight, zonulin expression and serum zonulin levels.
Materials and Methods: This laboratory experiment used 18 ddY mice, which were randomly divided into control (C), PBS (T1), and PBS + NS1 (T2) groups. Mice in the T1 and T2 groups were intravenously injected with 500 µl PBS only and 50 µg NS1 respectively. Mice blood samples were collected before and after three-day treatment for measurement of zonulin level. The fresh liver was weighted directly and were then used for immunostaining.
Results: The C group had lower wet liver weight compared to the T groups (p=0.001). Increased expression of liver zonulin was found in the T2 group, significant different from the C (p=0.014) and T1 groups (p=0.020). After treatment, serum zonulin levels in the T1 group was higher than that of the T1 group before treatment (p=0.035) but not in control (p=0.753) and T2 groups (p=0.869).
Conclusion: Administration of 50 µg NS 1 increases wet liver weight and zonulin expression in hepatocytes, but did not increase serum zonulin levels in ddY mice.
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2. Uno N, Ross TM. Dengue virus and the host innate immune response. Emerg Microbes Infect 2018; 7: 167.
3. Lin G-L, McGinley JP, Drysdale SB, Pollard AJ. Epidemiology and immune pathogenesis of viral sepsis. Front Immunol 2018; 9: 2147.
4. Harapan H, Michie A, Mudatsir M, Sasmono RT, Imrie A. Epidemiology of dengue hemorrhagic fever in Indonesia: analysis of five decades data from the National Disease Surveillance. BMC Res Notes 2019; 12: 350.
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6. Malavige GN, Ogg GS. Pathogenesis of vascular leak in dengue virus infection. Immunology 2017; 151: 261-269.
7. Chen H-R, Lai Y-C, Yeh T-M. Dengue virus non-structural protein 1: A pathogenic factor, therapeutic target, and vaccine candidate. J Biomed Sci 2018; 25: 58.
8. Zeidler JD, Fernandes-Siqueira LO, Barbosa GM, Da Poian AT. Non-canonical roles of dengue virus non-structural proteins. Viruses 2017; 9: 42.
9. Glasner DR, Puerta-Guardo H, Beatty PR, Harris E. The good, the bad, and the shocking: the multiple roles of dengue virus nonstructural protein 1 in protection and pathogenesis. Annu Rev Virol 2018; 5: 227-253.
10. Dwivedi VD, Tripathi IP, Tripathi RC, Bharadwaj S, Mishra SK. Genomics, proteomics and evolution of dengue virus. Brief Funct Genomics 2017; 16: 217-227.
11. Pang X, Zhang R, Cheng G. Progress towards understanding the pathogenesis of dengue hemorrhagic fever. Virol Sin 2017; 32: 16-22.
12. Kumar Y, Liang C, Bo Z, Rajapakse JC, Ooi EE, Tannenbaum SR. Serum proteome and cytokine analysis in a Longitudinal cohort of adults with primary Dengue infection reveals predictive Markers of DHF. PLoS Negl Trop Dis 2012; 6(11): e1887.
13. Perez L. Acute phase protein response to viral infection and vaccination. Arch Biochem Biophys 2019; 671: 196-202.
14. The UniProt Consortium. Haptoglobin. Geneva: UniProt Consortium; 2022. [cited 2022 August 3]. Available from: https://www.uniprot.org/uniprotkb/P00738/entry
15. Serek P, Oleksy-Wawrzyniak M. The Effect of Bacterial Infections, Probiotics and Zonulin on Intestinal Barrier Integrity. Int J Mol Sci 2021; 22: 11359.
16. Heinemann U, Schuetz A. Structural features of tight-junction proteins. Int J Mol Sci 2019; 20: 6020.
17. Assimakopoulos SF, Triantos C, Thomopoulos K, Fligou F, Maroulis I, Marangos M, et al. Gut-origin sepsis in the critically ill patient: pathophysiology and treatment. Infection 2018; 46: 751-760.
18. Giron LB, Dweep H, Yin X, Wang H, Damra M, Goldman AR, et al. Plasma markers of disrupted gut permeability in severe COVID-19 patients. Front Immunol 2021; 12: 686240.
19. Chancharoenthana W, Leelahavanichkul A, Ariyanon W, Vadcharavivad S, Phatcharophaswattanakul S, Kamolratanakul S, et al. Leaky gut syndrome is associated with endotoxemia and serum (1→3)-β-D-Glucan in severe dengue infection. Microorganisms 2021; 9: 2390.
20. Douglas KO, Samuels TA, Gittens-St Hilaire M. Serum LPS associated with Hantavirus and dengue disease severity in Barbados. Viruses 2019; 11: 838.
21. Zellweger RM, Shresta S. Mouse models to study dengue virus immunology and pathogenesis. Front Immunol 2014; 5: 151.
22. Linke A, Tiegs G, Neumann K. Pathogenic T-Cell Responses in Immune-Mediated Glomerulonephritis. Cells 2022; 11: 1625.
23. Arifin WN, Zahiruddin WM. Sample size calculation in animal studies using resource equation approach. Malays J Med Sci 2017; 24: 101-105.
24. Scudamore CL, Busk N, Vowell K. A simplified necropsy technique for mice: making the most of unscheduled deaths. Lab Anim 2014; 48: 342-344.
25. Fernando S, Wijewickrama A, Gomes L, Punchihewa CT, Madusanka SDP, Dissanayake H, et al. Patterns and causes of liver involvement in acute dengue infection. BMC Infect Dis 2016; 16: 319.
26. Lewis J, Mitra A, Chang M. Acute liver failure in a patient with dengue shock syndrome. ACG Case Rep J 2020; 7(4): e00371.
27. Suganthan N, Sakthilingham G, Kumanan T. Dengue fever complicated with acute liver failure: A case report of expanded dengue syndrome and literature review. SAGE Open Med Case Rep 2020; 8: 2050313X20913428.
28. Lee PX, Ting DHR, Boey CPH, Tan ETX, Chia JZH, Idris F, et al. Relative contribution of nonstructural protein 1 in dengue pathogenesis. J Exp Med 2020; 217(9): e20191548.
29. Rastogi M, Sharma N, Singh SK. Flavivirus NS1: a multifaceted enigmatic viral protein. Virol J 2016; 13: 131.
30. Chen H-R, Chuang Y-C, Lin Y-S, Liu H-S, Liu C-C, Perng G-C, et al. Dengue virus nonstructural protein 1 induces vascular Leakage through Macrophage Migration inhibitory factor and autophagy. PLoS Negl Trop Dis 2016; 10(7): e0004828.
31. Lin CY, Kolliopoulos C, Huang CH, Tenhunen J, Heldin CH, Chen YH, et al. High levels of serum hyaluronan is an early predictor of dengue warning signs and perturbs vascular integrity. EBioMedicine 2019; 48: 425-441.
32. Teo QW, van Leur SW, Sanyal S. Escaping the Lion's Den: redirecting autophagy for unconventional release and spread of viruses. FEBS J 2021; 288: 3913-3927.
33. Puerta-Guardo H, Glasner DR, Espinosa DA, Biering SB, Patana M, Ratnasiri K, et al. Flavivirus NS1 triggers tissue-specific vascular endothelial dysfunction reflecting disease Tropism. Cell Rep 2019; 26: 1598-1613.e8.
34. Puerta-Guardo H, Glasner DR, Harris E. Dengue virus NS1 disrupts the endothelial Glycocalyx, Leading to Hyperpermeability. PLoS Pathog 2016; 12(7): e1005738.
35. Silva EM, Conde JN, Allonso D, Nogueira ML, Mohana-Borges R. Mapping the interactions of dengue virus NS1 protein with human liver proteins using a yeast two-hybrid system: identification of C1q as an interacting partner. PLoS One 2013; 8(3): e57514.
36. Suwarto S, Sasmono RT, Sinto R, Ibrahim E, Suryamin M. Association of endothelial glycocalyx and tight and adherens junctions with severity of plasma leakage in dengue infection. J Infect Dis 2017; 215: 992–999.
37. Giang HTN, Banno K, Minh LHN, Trinh LT, Loc LT, Eltobgy A, et al. Dengue hemophagocytic syndrome: A systematic review and meta-analysis on epidemiology, clinical signs, outcomes, and risk factors. Rev Med Virol 2018; 28(6): e2005.
38. Leowattana W, Leowattana T. Dengue hemorrhagic fever and the liver. World J Hepatol 2021; 13: 1968-1976.
39. Alayli F, Scholle F. Dengue virus NS1 enhances viral replication and pro-inflammatory cytokine production in human dendritic cells. Virology 2016; 496: 227-236.
40. Benfrid S, Park K-H, Dellarole M, Voss JE, Tamietti C, Pehau-Arnaudet G, et al. Dengue virus NS1 protein conveys pro-inflammatory signals by docking onto high-density lipoproteins. EMBO Rep 2022; 23(7): e53600.
41. Khalil RH, Al-Humadi N. Types of acute phase reactants and their importance in vaccination. Biomed Rep 2020; 12: 143-152.
42. Soe HJ, Manikam R, Raju CS, Khan MA, Sekaran SD. Correlation of host inflammatory cytokines and immune-related metabolites, but not viral NS1 protein, with disease severity of dengue virus infection. PLoS One 2020; 15(8): e0237141.
43. Hartono, Suryawati B, Sari Y, Avicena A, Maryani, Sukmagautama C, et al. The effect of curcumin and virgin coconut oil towards cytokines levels in COVID-19 patients at universitas Sebelas Maret Hospital, Surakarta, Indonesia. Pharmacogn J 2022; 14: 216-225.
44. Akao T, Morita A, Onji M, Miyake T, Watanabe R, Uehara T, et al. Low serum levels of Zonulin in patients with HCV-infected chronic liver diseases. Euroasian J Hepatogastroenterol 2018; 8: 112-115.
45. Wolf-Grosse S, Rokstad AM, Ali S, Lambris JD, Mollnes TE, Nilsen AM, et al. Iron oxide nanoparticles induce cytokine secretion in a complement-dependent manner in a human whole blood model. Int J Nanomedicine 2017; 12: 3927-3940.
46. Wang Y, Zhou H, Shen Y, Wang Y, Wu W, Liu H, et al. Impairment of dendritic cell function and induction of CD4(+) CD25(+) Foxp 3(+) T cells by excretory-secretory products: a potential mechanism of immune evasion adopted by Echinococcus granulosus. BMC Immunol 2015; 16: 44.
47. Liang SC, Nickerson-Nutter C, Pittman DD, Carrier Y, Goodwin DG, Shields KM, et al. IL-22 Induces an acute-phase response. J Immunol 2010; 185: 5531-5538.
48. Saxton RA, Henneberg LT, Calafiore M, Su L, Jude KM, Hanash AM, et al. The tissue protective functions of interleukin-22 can be decoupled from pro-inflammatory actions through structure-based design. Immunity 2021; 54: 660-672.e9.
49. Doolittle JM, Gomez SM. Mapping protein interactions between Dengue Virus and its human and insect hosts. PLoS Negl Trop Dis 2011; 5(2): e954.
2. Uno N, Ross TM. Dengue virus and the host innate immune response. Emerg Microbes Infect 2018; 7: 167.
3. Lin G-L, McGinley JP, Drysdale SB, Pollard AJ. Epidemiology and immune pathogenesis of viral sepsis. Front Immunol 2018; 9: 2147.
4. Harapan H, Michie A, Mudatsir M, Sasmono RT, Imrie A. Epidemiology of dengue hemorrhagic fever in Indonesia: analysis of five decades data from the National Disease Surveillance. BMC Res Notes 2019; 12: 350.
5. Lee J-C, Cia C-T, Lee N-Y, Ko N-Y, Chen P-L, Ko W-C. Causes of death among dengue patients causes of death among hospitalized adults with dengue fever in Tainan, 2015: Emphasis on cardiac events and bacterial infections. J Microbiol Immunol Infect 2022; 55: 207-214.
6. Malavige GN, Ogg GS. Pathogenesis of vascular leak in dengue virus infection. Immunology 2017; 151: 261-269.
7. Chen H-R, Lai Y-C, Yeh T-M. Dengue virus non-structural protein 1: A pathogenic factor, therapeutic target, and vaccine candidate. J Biomed Sci 2018; 25: 58.
8. Zeidler JD, Fernandes-Siqueira LO, Barbosa GM, Da Poian AT. Non-canonical roles of dengue virus non-structural proteins. Viruses 2017; 9: 42.
9. Glasner DR, Puerta-Guardo H, Beatty PR, Harris E. The good, the bad, and the shocking: the multiple roles of dengue virus nonstructural protein 1 in protection and pathogenesis. Annu Rev Virol 2018; 5: 227-253.
10. Dwivedi VD, Tripathi IP, Tripathi RC, Bharadwaj S, Mishra SK. Genomics, proteomics and evolution of dengue virus. Brief Funct Genomics 2017; 16: 217-227.
11. Pang X, Zhang R, Cheng G. Progress towards understanding the pathogenesis of dengue hemorrhagic fever. Virol Sin 2017; 32: 16-22.
12. Kumar Y, Liang C, Bo Z, Rajapakse JC, Ooi EE, Tannenbaum SR. Serum proteome and cytokine analysis in a Longitudinal cohort of adults with primary Dengue infection reveals predictive Markers of DHF. PLoS Negl Trop Dis 2012; 6(11): e1887.
13. Perez L. Acute phase protein response to viral infection and vaccination. Arch Biochem Biophys 2019; 671: 196-202.
14. The UniProt Consortium. Haptoglobin. Geneva: UniProt Consortium; 2022. [cited 2022 August 3]. Available from: https://www.uniprot.org/uniprotkb/P00738/entry
15. Serek P, Oleksy-Wawrzyniak M. The Effect of Bacterial Infections, Probiotics and Zonulin on Intestinal Barrier Integrity. Int J Mol Sci 2021; 22: 11359.
16. Heinemann U, Schuetz A. Structural features of tight-junction proteins. Int J Mol Sci 2019; 20: 6020.
17. Assimakopoulos SF, Triantos C, Thomopoulos K, Fligou F, Maroulis I, Marangos M, et al. Gut-origin sepsis in the critically ill patient: pathophysiology and treatment. Infection 2018; 46: 751-760.
18. Giron LB, Dweep H, Yin X, Wang H, Damra M, Goldman AR, et al. Plasma markers of disrupted gut permeability in severe COVID-19 patients. Front Immunol 2021; 12: 686240.
19. Chancharoenthana W, Leelahavanichkul A, Ariyanon W, Vadcharavivad S, Phatcharophaswattanakul S, Kamolratanakul S, et al. Leaky gut syndrome is associated with endotoxemia and serum (1→3)-β-D-Glucan in severe dengue infection. Microorganisms 2021; 9: 2390.
20. Douglas KO, Samuels TA, Gittens-St Hilaire M. Serum LPS associated with Hantavirus and dengue disease severity in Barbados. Viruses 2019; 11: 838.
21. Zellweger RM, Shresta S. Mouse models to study dengue virus immunology and pathogenesis. Front Immunol 2014; 5: 151.
22. Linke A, Tiegs G, Neumann K. Pathogenic T-Cell Responses in Immune-Mediated Glomerulonephritis. Cells 2022; 11: 1625.
23. Arifin WN, Zahiruddin WM. Sample size calculation in animal studies using resource equation approach. Malays J Med Sci 2017; 24: 101-105.
24. Scudamore CL, Busk N, Vowell K. A simplified necropsy technique for mice: making the most of unscheduled deaths. Lab Anim 2014; 48: 342-344.
25. Fernando S, Wijewickrama A, Gomes L, Punchihewa CT, Madusanka SDP, Dissanayake H, et al. Patterns and causes of liver involvement in acute dengue infection. BMC Infect Dis 2016; 16: 319.
26. Lewis J, Mitra A, Chang M. Acute liver failure in a patient with dengue shock syndrome. ACG Case Rep J 2020; 7(4): e00371.
27. Suganthan N, Sakthilingham G, Kumanan T. Dengue fever complicated with acute liver failure: A case report of expanded dengue syndrome and literature review. SAGE Open Med Case Rep 2020; 8: 2050313X20913428.
28. Lee PX, Ting DHR, Boey CPH, Tan ETX, Chia JZH, Idris F, et al. Relative contribution of nonstructural protein 1 in dengue pathogenesis. J Exp Med 2020; 217(9): e20191548.
29. Rastogi M, Sharma N, Singh SK. Flavivirus NS1: a multifaceted enigmatic viral protein. Virol J 2016; 13: 131.
30. Chen H-R, Chuang Y-C, Lin Y-S, Liu H-S, Liu C-C, Perng G-C, et al. Dengue virus nonstructural protein 1 induces vascular Leakage through Macrophage Migration inhibitory factor and autophagy. PLoS Negl Trop Dis 2016; 10(7): e0004828.
31. Lin CY, Kolliopoulos C, Huang CH, Tenhunen J, Heldin CH, Chen YH, et al. High levels of serum hyaluronan is an early predictor of dengue warning signs and perturbs vascular integrity. EBioMedicine 2019; 48: 425-441.
32. Teo QW, van Leur SW, Sanyal S. Escaping the Lion's Den: redirecting autophagy for unconventional release and spread of viruses. FEBS J 2021; 288: 3913-3927.
33. Puerta-Guardo H, Glasner DR, Espinosa DA, Biering SB, Patana M, Ratnasiri K, et al. Flavivirus NS1 triggers tissue-specific vascular endothelial dysfunction reflecting disease Tropism. Cell Rep 2019; 26: 1598-1613.e8.
34. Puerta-Guardo H, Glasner DR, Harris E. Dengue virus NS1 disrupts the endothelial Glycocalyx, Leading to Hyperpermeability. PLoS Pathog 2016; 12(7): e1005738.
35. Silva EM, Conde JN, Allonso D, Nogueira ML, Mohana-Borges R. Mapping the interactions of dengue virus NS1 protein with human liver proteins using a yeast two-hybrid system: identification of C1q as an interacting partner. PLoS One 2013; 8(3): e57514.
36. Suwarto S, Sasmono RT, Sinto R, Ibrahim E, Suryamin M. Association of endothelial glycocalyx and tight and adherens junctions with severity of plasma leakage in dengue infection. J Infect Dis 2017; 215: 992–999.
37. Giang HTN, Banno K, Minh LHN, Trinh LT, Loc LT, Eltobgy A, et al. Dengue hemophagocytic syndrome: A systematic review and meta-analysis on epidemiology, clinical signs, outcomes, and risk factors. Rev Med Virol 2018; 28(6): e2005.
38. Leowattana W, Leowattana T. Dengue hemorrhagic fever and the liver. World J Hepatol 2021; 13: 1968-1976.
39. Alayli F, Scholle F. Dengue virus NS1 enhances viral replication and pro-inflammatory cytokine production in human dendritic cells. Virology 2016; 496: 227-236.
40. Benfrid S, Park K-H, Dellarole M, Voss JE, Tamietti C, Pehau-Arnaudet G, et al. Dengue virus NS1 protein conveys pro-inflammatory signals by docking onto high-density lipoproteins. EMBO Rep 2022; 23(7): e53600.
41. Khalil RH, Al-Humadi N. Types of acute phase reactants and their importance in vaccination. Biomed Rep 2020; 12: 143-152.
42. Soe HJ, Manikam R, Raju CS, Khan MA, Sekaran SD. Correlation of host inflammatory cytokines and immune-related metabolites, but not viral NS1 protein, with disease severity of dengue virus infection. PLoS One 2020; 15(8): e0237141.
43. Hartono, Suryawati B, Sari Y, Avicena A, Maryani, Sukmagautama C, et al. The effect of curcumin and virgin coconut oil towards cytokines levels in COVID-19 patients at universitas Sebelas Maret Hospital, Surakarta, Indonesia. Pharmacogn J 2022; 14: 216-225.
44. Akao T, Morita A, Onji M, Miyake T, Watanabe R, Uehara T, et al. Low serum levels of Zonulin in patients with HCV-infected chronic liver diseases. Euroasian J Hepatogastroenterol 2018; 8: 112-115.
45. Wolf-Grosse S, Rokstad AM, Ali S, Lambris JD, Mollnes TE, Nilsen AM, et al. Iron oxide nanoparticles induce cytokine secretion in a complement-dependent manner in a human whole blood model. Int J Nanomedicine 2017; 12: 3927-3940.
46. Wang Y, Zhou H, Shen Y, Wang Y, Wu W, Liu H, et al. Impairment of dendritic cell function and induction of CD4(+) CD25(+) Foxp 3(+) T cells by excretory-secretory products: a potential mechanism of immune evasion adopted by Echinococcus granulosus. BMC Immunol 2015; 16: 44.
47. Liang SC, Nickerson-Nutter C, Pittman DD, Carrier Y, Goodwin DG, Shields KM, et al. IL-22 Induces an acute-phase response. J Immunol 2010; 185: 5531-5538.
48. Saxton RA, Henneberg LT, Calafiore M, Su L, Jude KM, Hanash AM, et al. The tissue protective functions of interleukin-22 can be decoupled from pro-inflammatory actions through structure-based design. Immunity 2021; 54: 660-672.e9.
49. Doolittle JM, Gomez SM. Mapping protein interactions between Dengue Virus and its human and insect hosts. PLoS Negl Trop Dis 2011; 5(2): e954.
| Files | ||
| Issue | Vol 15 No 2 (2023) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v15i2.12483 | |
| Keywords | ||
| Acute phase protein; Dengue virus type 2; Hepatomegaly; Nonstructural protein 1; Zonulin | ||
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How to Cite
1.
Jatmiko S, Hartono H, Ardyanto T, Indarto D. The acute effects of nonstructural-1 protein dengue virus type 2 on wet liver weight, zonulin expression and serum zonulin. Iran J Microbiol. 2023;15(2):311-317.
