The Immunohistochemical and Bioinformatics Analysis of the Placental Expressions of Vascular Cell Adhesion Protein 1 (VCAM-1) and High Mobility Group Box 1 (HMGB1) Proteins in Gestational Diabetic Mothers

Abstract

Objective  We aimed to examine both the expression levels of high mobility group box 1 (HMGB1) and vascular cell adhesion molecule-1 (VCAM-1) proteins in the placentas of pregnant women with gestational diabetes mellitus (GDM) and control groups by immunohistochemical (IHC) method.

Material and methods  An experimental case-control study was conducted, including 40 pregnant women complicated with GDM and 40 healthy pregnant women. Placental tissues obtained following cesarean delivery were subjected to routine tissue monitoring. The placental sections were stained with VCAM-1 and HMGB1 immunostains and subjected to IHC examination under a light microscope. H-score (HS) was used to evaluate the results of IHC staining by semi-quantitative analysis. Pathway analysis in Cytoscape software identified GDM-associated proteins within HMGB1 and VCAM-1 interaction networks, followed by GO analysis to explore associated biological processes.

Results  Placental HGMB1 expression was significantly increased in the GDM group compared to the control group (p<0.001). However, placental VCAM-1 expression was found to be statistically similar in GDM and control groups (p=0.584). The shared 19 proteins were identified between HMGB1 and GDM, and 13 between VCAM-1 and GDM, with notable GO biological process terms such as immune system activation for HMGB1 and interleukin-6 regulation for VCAM-1 associated with GDM.

Conclusion  We consider that GDM-related inflammation and oxidative stress may contribute to tissue damage and inflammation by increasing placental HMGB1 expression. The blockade of HMGB1 and its receptors might represent a promising therapeutic approach to control inflammation in GDM. Understanding the distinct roles of HMGB1 and VCAM-1 may provide valuable insights for the development of targeted therapies aimed at mitigating the inflammatory processes associated with GDM and improving maternal and fetal outcomes.

Keywords

gestational dabetes mellitus - high mobility group box 1 - vascular cell adhesion molecule-1 - immunohistochemical analysis

Publication History

Received: 09 April 2024

Accepted after revision: 06 October 2024

Article published online:
12 November 2024

• References

• 1 American Diabetes Association Professional Practice Committee. 2. Classification and Diagnosis of Diabetes: Standards of Medical Care in Diabetes – 2022. Diabetes Care 2022; 45: S17-S38
• 2 Sacks DA, Hadden DR, Maresh M. et al. HAPO Study Cooperative Research Group. Frequency of gestational diabetes mellitus at collaborating centers based on IADPSG consensus panel-recommended criteria: the Hyperglycemia and Adverse Pregnancy Outcome (HAPO) Study. Diabetes Care 2012; 35: 526-528
• 3 Chiefari E, Arcidiacono B, Foti D, Brunetti A. Gestational diabetes mellitus: an updated overview. J Endocrinol Invest 2017; 40: 899-909
• 4 Oğlak SC, Yavuz A, Olmez F. et al. The reduced serum concentrations of β-arrestin-1 and β-arrestin-2 in pregnancies complicated with gestational diabetes mellitus. J Matern Fetal Neonatal Med 2022; 35: 10017-10024
• 5 Akgöl S, Budak MŞ, Oğlak SC. et al. Can maternal abdominal fat thickness predict antenatal insulin therapy in patients with gestational diabetes mellitus?. J Obstet Gynaecol Res 2022; 48: 634-639
• 6 Nakshine VS, Jogdand SD. A comprehensive review of gestational diabetes mellitus: Impacts on maternal health, fetal development, childhood outcomes, and long-term treatment strategies. Cureus 2023; 15: e47500
• 7 Tunc S, Oglak SC, Olmez F, Ozkose ZG. The value of first-trimester maternal abdominal visceral adipose tissue thickness in predicting the subsequent development of gestational diabetes mellitus. J Coll Physicians Surg Pak 2022; 32: 722-727
• 8 Aldahmash WM, Alwasel SH, Aljerian K. Gestational diabetes mellitus induces placental vasculopathies. Environ Sci Pollut Res Int 2022; 29: 19860-19868
• 9 Scifres CM, Parks WT, Feghali M. et al. Placental maternal vascular malperfusion and adverse pregnancy outcomes in gestational diabetes mellitus. Placenta 2017; 49: 10-15
• 10 Oğlak SC, Obut M. Expression of ADAMTS13 and PCNA in the placentas of gestational diabetic mothers. Int J Morphol 2021; 39: 38-44
• 11 Edu A, Teodorescu C, Dobjanschi CG. et al. Placenta changes in pregnancy with gestational diabetes. Rom J Morphol Embryol 2016; 57: 507-512
• 12 Holmlund U, Wähämaa H, Bachmayer N. et al. The novel inflammatory cytokine high mobility group box protein 1 (HMGB1) is expressed by human term placenta. Immunology 2007; 122: 430-437
• 13 Wu H, Li R, Wei ZH. et al. Diabetes-induced oxidative stress in endothelial progenitor cells may be sustained by a positive feedback loop involving high mobility group box-1. Oxid Med Cell Longev 2016; 2016: 1943918
• 14 Andersson U, Tracey KJ. HMGB1 is a therapeutic target for sterile inflammation and infection. Annu Rev Immunol 2011; 29: 139-162
• 15 Yang K, Cao F, Wang W. et al. The relationship between HMGB1 and autophagy in the pathogenesis of diabetes and its complications. Front Endocrinol (Lausanne) 2023; 14: 1141516
• 16 Nogueira-Machado JA, Volpe CM, Veloso CA, Chaves MM. HMGB1, TLR and RAGE: a functional tripod that leads to diabetic inflammation. Expert Opin Ther Targets 2011; 15: 1023-1035
• 17 Cook-Mills JM, Marchese ME, Abdala-Valencia H. Vascular cell adhesion molecule-1 expression and signaling during disease: regulation by reactive oxygen species and antioxidants. Antioxid Redox Signal 2011; 15: 1607-1638
• 18 Kong DH, Kim YK, Kim MR. et al. Emerging roles of vascular cell adhesion molecule-1 (VCAM-1) in immunological disorders and cancer. Int J Mol Sci 2018; 19: 1057
• 19 International Association of Diabetes and Pregnancy Study Groups Consensus Panel. Metzger BE, Gabbe SG, Persson B. et al. International association of diabetes and pregnancy study groups recommendations on the diagnosis and classification of hyperglycemia in pregnancy. Diabetes Care 2010; 33: 676-682
• 20 Cizkova K, Foltynkova T, Gachechiladze M, Tauber Z. Comparative Analysis of Immunohistochemical Staining Intensity Determined by Light Microscopy, ImageJ and QuPath in Placental Hofbauer Cells. Acta Histochem Cytochem 2021; 54: 21-29
• 21 Ge SX, Jung D, Yao R. ShinyGO: a graphical gene-set enrichment tool for animals and plants. Bioinformatics 2020; 36: 2628-2629
• 22 Johns EC, Denison FC, Norman JE, Reynolds RM. Gestational diabetes mellitus: Mechanisms, treatment, and complications. Trends Endocrinol Metab 2018; 29: 743-754
• 23 Šimják P, Cinkajzlová A, Anderlová K. et al. The role of obesity and adipose tissue dysfunction in gestational diabetes mellitus. J Endocrinol 2018; 238: R63-R77
• 24 Plows JF, Stanley JL, Baker PN. et al. The pathophysiology of gestational diabetes mellitus. Int J Mol Sci 2018; 19: 3342
• 25 Zhu Y, Zhang C. Prevalence of gestational diabetes and risk of progression to Type 2 diabetes: a global perspective. Curr Diab Rep 2016; 16: 7
• 26 Asir F, Oglak SC, Korak T. et al. Placental vimentin expression in preeclampsia and gestational diabetes mellitus. Gynecol Obstet Reprod Med 2024; 30: 10-18
• 27 Gebes O, Kale İbrahim, Beser Gebes T, Muhcu M. Investigation of serum cartonectin concentrations in pregnant women with gestational diabetes mellitus; a prospective non-interventional cohort study. Gynecol Obstet Reprod Med. 2024 30. 33-38
• 28 Holmlund U, Wähämaa H, Bachmayer N. et al. The novel inflammatory cytokine high mobility group box protein 1 (HMGB1) is expressed by human term placenta. Immunology 2007; 122: 430-437
• 29 Zenerino C, Nuzzo AM, Giuffrida D. et al. The HMGB1/rage pro-inflammatory axis in the human placenta: Modulating effect of low molecular weight heparin. Molecules 2017; 22: 1997
• 30 Wang Y, Zhong J, Zhang X. et al. The role of HMGB1 in the pathogenesis of Type 2 diabetes. J Diabetes Res 2016; 2016: 2543268
• 31 Pachydaki SI, Tari SR, Lee SE. et al. Upregulation of RAGE and its ligands in proliferative retinal disease. Exp Eye Res 2006; 82: 807-815
• 32 Yu Y, Yang L, Lv J. et al. The role of high mobility group box 1 (HMGB-1) in the diabetic retinopathy inflammation and apoptosis. Int J Clin Exp Pathol 2015; 8: 6807-6813
• 33 Dasu MR, Devaraj S, Park S, Jialal I. Increased toll-like receptor (TLR) activation and TLR ligands in recently diagnosed type 2 diabetic subjects. Diabetes Care 2010; 33: 861-868
• 34 Skrha J, Kalousová M, Svarcová J. et al. Relationship of soluble RAGE and RAGE ligands HMGB1 and EN-RAGE to endothelial dysfunction in type 1 and type 2 diabetes mellitus. Exp Clin Endocrinol Diabetes 2012; 120: 277-281
• 35 Giacobbe A, Granese R, Grasso R. et al. Association between maternal serum high mobility group box 1 levels and pregnancy complicated by gestational diabetes mellitus. Nutr Metab Cardiovasc Dis 2016; 26: 414-418
• 36 Hagiwara S, Iwasaka H, Hasegawa A. et al. Effects of hyperglycemia and insulin therapy on high mobility group box 1 in endotoxin-induced acute lung injury in a rat model. Crit Care Med 2008; 36: 2407-2413
• 37 Tsoyi K, Jang HJ, Nizamutdinova IT. et al. Metformin inhibits HMGB1 release in LPS-treated RAW 264.7 cells and increases survival rate of endotoxaemic mice. Br J Pharmacol 2011; 162: 1498-1508
• 38 Zhang T, Hu X, Cai Y. et al. Metformin protects against hyperglycemia-induced cardiomyocytes injury by inhibiting the expressions of receptor for advanced glycation end products and high mobility group box 1 protein. Mol Biol Rep 2014; 41: 1335-1340
• 39 Klune JR, Dhupar R, Cardinal J. et al. HMGB1: endogenous danger signaling. Mol Med 2008; 14: 476-484
• 40 Xu H, Barnes GT, Yang Q. et al. Chronic inflammation in fat plays a crucial role in the development of obesity-related insulin resistance. J Clin Invest 2003; 112: 1821-1830
• 41 Yang H, Wang H, Czura CJ, Tracey KJ. The cytokine activity of HMGB1. J Leukoc Biol 2005; 78: 1-8
• 42 Burlina S, Dalfrà MG, Chilelli NC, Lapolla A. Gestational diabetes mellitus and future cardiovascular risk: an update. Int J Endocrinol 2016; 2016: 2070926
• 43 Göbl CS, Bozkurt L, Yarragudi R. et al. Biomarkers of endothelial dysfunction in relation to impaired carbohydrate metabolism following pregnancy with gestational diabetes mellitus. Cardiovasc Diabetol 2014; 13: 138
• 44 Kautzky-Willer A, Fasching P, Jilma B. et al. Persistent elevation and metabolic dependence of circulating E-selectin after delivery in women with gestational diabetes mellitus. J Clin Endocrinol Metab 1997; 82: 4117-4121
• 45 Siddiqui K, George TP, Nawaz SS, Joy SS. VCAM-1, ICAM-1 and selectins in gestational diabetes mellitus and the risk for vascular disorders. Future Cardiol 2019; 15: 339-346
• 46 Loukeris K, Sela R, Baergen RN. Syncytial knots as a reflection of placental maturity: reference values for 20 to 40 weeksʼ gestational age. Pediatr Dev Pathol 2010; 13: 305-309
• 47 Fogarty NM, Ferguson-Smith AC, Burton GJ. Syncytial knots (Tenney-Parker changes) in the human placenta: evidence of loss of transcriptional activity and oxidative damage. Am J Pathol 2013; 183: 144-152
• 48 Olmos-Ortiz A, Flores-Espinosa P, Díaz L. et al. Immunoendocrine dysregulation during gestational diabetes mellitus: the central role of the placenta. Int J Mol Sci 2021; 22: 8087
• 49 Xuan Nguyen K, Bui Minh T, Dinh HT. et al. Low-grade inflammation in gestational diabetes mellitus and its correlation with maternal insulin resistance and fetal growth indices. Int J Gen Med 2023; 16: 1429-1436
• 50 Pantham P, Aye IL, Powell TL. Inflammation in maternal obesity and gestational diabetes mellitus. Placenta 2015; 36: 709-715
• 51 Rehman K, Akash MSH, Liaqat A. et al. Role of interleukin-6 in development of insulin resistance and type 2 diabetes mellitus. Crit Rev Eukaryot Gene Expr 2017; 27: 229-236
• 52 Amirian A, Mahani MB, Abdi F. Role of interleukin-6 (IL-6) in predicting gestational diabetes mellitus. Obstet Gynecol Sci 2020; 63: 407-416