Tag Archives: VEGF
Therapeutic potential of topical autologous angiostatin application in managing tuberculosis-related corneal injury: a case report
N. Greben1*, I. Gavryliak1, V. Bilous2,
V. Korsa2, A. Tykhomyrov2
1Department of Ophthalmology, Bogomolets National Medical University, Kyiv, Ukraine;
2Department of Enzyme Chemistry and Biochemistry,
Palladin Institute of Biochemistry, Kyiv, Ukraine;
*e-mail: nkgreden@ukr.net
Received: 03 June 2025; Revised: 17 September 2025;
Accepted: 28 November 2025; Available on-line: 29 December 2025
Ocular tuberculosis (TB) is a vision-threatening condition that frequently manifests as corneal neovascularization and stromal keratitis, which triggers a cascade of inflammatory and hypoxia-driven responses. Conventional therapeutic approaches, including corticosteroids and antimicrobial agents, often fail to halt disease progression. Here, we report a case of a 50-year-old patient diagnosed with TB-associated keratitis, unresponsive to standard treatment. The aim of the study was to evaluate the effectiveness of the alternative therapeutic strategy involving topical administration of angiostatin, a natural anti-angiogenic polypeptide derived from the autologous plasminogen. Solution of angiostatin fragment containing the first three kringle domains (K1-3) was applied in a two doses of eye drops (~15 μg per administration) five times daily for 2 months, with a cumulative exposure of approximately 4.5 mg. Treatment efficacy was monitored using both standard ophthalmologic assessments and non-invasive biochemical indicators such as the levels of hypoxia-inducible factor HIF-1α, vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP-9), fibrinogen/fibrin (Fg/Fb) and lactoferrin measured the in tear fluid across treatment time points (Day 0, 14, and 61) using Western blot analysis. The high intensity of HIF-1 α, VEGF and MMP-9 expression, Fg/Fb accumulation and the presence of low-molecular-weight fragments of lactoferrin were detected in the tear fluid prior to the treatment. Following angiostatin therapy, the patient exhibited marked regression of corneal neovascularization and restoration of corneal transparency, complemented with normalization of HIF-1α, VEGF, and MMP-9 levels, reduced Fg/Fb accumulation and the presence of intact lactoferrin in the tear fluid. The data obtained demonstrated a multifactorial mechanism of angiostatin action that extends beyond classical anti-angiogenic pathways. The convergence of clinical and molecular indicators of recovery underscores the potential of angiostatin application as a safe and effective therapeutic alternative for managing corneal complications in ocular TB, particularly in cases resistant to conventional treatment.
Circulating levels of potential markers of ischemic stroke in patients with the different forms of atrial fibrillation and chronic heart failure
A. O. Tykhomyrov1*, O. Yu. Sirenko2, O. V. Kuryata2
1Department of Enzyme Chemistry and Biochemistry,
Palladin Institute of Biochemistry, National Academy of Sciences of Ukraine, Kyiv, Ukraine;
2Department of Internal Medicine 2, Phthisiology,
Occupational Diseases and Clinical Immunology, Dnipro State Medical University, Dnipro, Ukraine;
*e-mail: artem_tykhomyrov@ukr.net
Received: 19 January 2024; Revised: 13 March 2024;
Accepted: 17 March 2024; Available on-line: 30 April 2024
Atrial fibrillation (AF) is the most common abnormal type of heart rhythm (cardiac arrhythmia), which is considered the leading cause of stroke. There have been limited studies on the prognostic markers for atrial disease and AF-associated ischemic stroke, despite the high demand for this procedure in daily clinical practice to monitor disease course and assess risk of stroke in patients with AF and chronic heart failure (CHF). Thus, the aim of the present study was to evaluate the levels of serum biomarkers related to ischemic stroke in CHF patients with the different forms of AF. Forty-six patients with various types of AF (paroxysmal, persistent and permanent) with or without ischemic stroke were enrolled in the study, 36 clinically healthy donors served as a control. The levels of inducible nitric oxide synthase (iNOS), vascular endothelial growth factor (VEGF) and angiostatins (AS) were evaluated by western blot analysis in the serum. The levels of active matrix metalloproteinases (MMPs) were analysed by gelatin zymography. Elevated levels of iNOS were shown in patients with all AF forms as compared with control, but iNOS levels in post-ischemic patients were significantly higher than that in paroxysmal AF individuals. However, the levels of VEGF and AS did not differ from the baseline value in patients with paroxysmal AF, while dramatic increase of their contents was shown in post-stroke patients with persistent and permanent types of AF. Elevated active MMP-9 levels were shown to be associated with the diagnosis of all AF forms, regardless of the occurrence of stroke. Taken together, our findings demonstrate that tested proteins can be considered as valuable biomarkers of AF forms transformation and potentially useful for ischemic stroke risk stratification in patients with AF and CHF. Observed changes in regulatory protein levels may expand our understanding of pathological roles of endothelial function dysregulation, disrupted angiogenesis balance and abnormal tissue remodeling in AF and associated ischemic events.
Multiple effects of angiostatins in injured cornea
V. L. Bilous*, A. O. Tykhomyrov
Department of Enzyme Chemistry and Biochemistry, Palladin Institute of Biochemistry,
National Academy of Sciences of Ukraine, Kyiv;
*e-mail: basil.bilous@gmail.com
Received: 07 October 2023; Revised: 01 November 2023;
Accepted: 01 February 2024; Available on-line: 26 February 2024
Prolonged inflammation and excessive neovascularization of the cornea due to severe injury can impair optical clarity and lead to vision impairment. Plasminogen kringle (K) fragments, known as angiostatins (AS), play a well-established role as inhibitors of neovascularization by suppressing pro-angiogenic signaling. However, AS effects in the cornea, beyond inhibiting the angiogenesis, are still unexplored. In this study, we estimate the protective effect of two AS variants (K1-3 and K5) against alkali burn injury induced in rabbit and rat corneas. AS K1-3 in the single doses of 0.075 or 0.75 μg (0.1 or 1.0 μM, respectively) or 0.3 μg of AS K5 (1.0 μM) were applied locally as eye drops daily for 14 days after the injury. A significant regression of corneal vessels in-growth in injured eyes treated with AS was revealed. Western blot analysis of corneal tissue lysates revealed that injury-induced overexpression of protein markers of hypoxia (HIF-1α), angiogenesis (VEGF), tissue remodeling and fibrosis (MMP-9), autophagy (beclin-1) and endoplasmic reticulum stress (GRP-78) was significantly reduced under AS treatment. Besides, the level of tight junctions protein ZO-1 was shown to be up-regulated after the treatment of the damaged cornea with AS K1-3. Summarizing, our study uncovered novel biological functions of the kringle-containing plasminogen fragments indicating its beneficial effects during corneal healing in the experimental model of alkali burn. The data obtained can be helpful for the development of novel efficient formulations to manage complications of ocular surface injuries.
Phenobarbital ameliorates hyperglycemia-induced angiogenesis in diabetic nephropathy-possible intervention at the HIF-1α/VEGF axis
M. M. Mohammed1*, S. R. A. Rehim1, A. M. M. Okasha1, H. El-Mezayen2,
D. G. A. N. Mohammed2, W. Gomaa3, F. Mourad4, E. G. Ayad2
1Department of Biochemistry, Faculty of Medicine, Minia University, El-Minia, Egypt;
2Department of Chemistry, Faculty of Science, Helwan University, Helwan, Egypt;
3Department of Pathology, Faculty of Medicine, Minia University, El-Minia, Egypt;
4MSP, Faculty of Pharmacy, Deraya University, El-Minia, Egypt;
*e-mail: mostafa.mohamed@mu.edu.eg
Received: 17 September 2023; Revised: 17 November 2023;
Accepted: 01 December 2023; Available on-line: 18 December 2023
Hyperglycemia contributes to a cascade of inflammatory responses in kidneys that result in the development of renal hypoxia and angiogenesis with subsequent chronic renal failure. As the hypoxia-inducible factor-1α (HIF-1α)/vascular endothelial growth factor (VEGF) axis is a key pathway for neovascularization, the inhibition of this axis is a target for renal angiogenesis therapy. We speculate that Phenobarbital (PB) which has a potential to reduce vascularization in clinical settings might have an influence on the development of angiogenesis in diabetic kidney. The aim of the study was to explore the effects of PB on the HIF-1α and VEGF expression and angiogenesis in renal tissue of rats with hyperglycemia and diabetic nephropathy. Sixty-four male Wistar rats were devided into 4 groups: control group received a single intraperitoneal saline injection; PB group received 0.05% PB orally in drinking water; diabetic group received a single intra-peritoneal STZ (65 mg/kg) injection; PB-STZ group received 0.05% PB orally two weeks before STZ administration. At the end of the experiment period (8 weeks), the kidneys were removed and used for biochemical analyses. Serum glucose, urea and creatinine levels, IL-6 levels in kidney homogenate and changes in HIF-1α and VEGF gene expression were estimated. Hematoxylin-eosin staining was performed for histopathological examination. The results obtained showed that both HIF-1α and VEGF gene expression and IL6 level in diabetic rat group were significantly elevated compared to that in control group, whereas in PB and PB-STZ groups, these indices were significantly down-regulated compared to the diabetic group. Abundant glomerular congestion and mesangial proliferation were detected in diabetic rat renal tissues. However, in PB-treated diabetic group, newly formed vessels were significantly decreased. These findings confirmed that phenobarbital, affecting the HIF-1α/VEGF signaling pathway, ameliorates angiogenesis after hyperglycemic kidney injury.
Benzodiazepine receptor agonist carbacetam modulates the level of vascular endothelial growth factor in the retina of rats with streptozotocin-induced diabetes
S. V. Ziablitsev1*, D. B. Zhupan1, A. O. Tykhomyrov2, O. O. Dyadyk3
1Bogomolets National Medical University, Kyiv, Ukraine;
2Palladin Institute of Biochemistry, National Academy
of Sciences of Ukraine, Kyiv, Ukraine;
3Shupyk National Healthcare University of Ukraine, Kyiv, Ukraine;
*e-mail: zsv1965@gmail.com
Received: 07 September 2023; Revised: 19 October 2023;
Accepted: 01 December 2023; Available on-line: 18 December 2023
One of the primary mechanisms of retinal neurodegeneration in diabetes mellitus is gamma-aminobutyric acid (GABA) deficiency that makes the use of GABA-benzodiazepine receptor modulators a promising option for the correction of this diabetic complication. The aim of this study was to determine the effect of the benzodiazepine receptor agonist carbacetam on the expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor-1α (HIF-1α) in retina of rats with hyperglycemia. Experimental diabetes was modeled by a single administration of streptozotocin (50 mg/kg) to three-month-old male Wistar rats. Immunoblotting and immunohistochemical studies were performed using monoclonal antibodies against VEGF and HIF-1α. It was shown that the development of diabetic retinopathy (DR) at the early stages was accompanied by a progressive multifold increase in the retina content of VEGF on 7-28 days and HIF-1α on 28th day. Insulin and insulin+carbacetam treatment significantly alleviated diabetes-induced overexpression of both HIF-1α and VEGF. Carbacetam was shown to block the diabetogenic increase in VEGF content in retina. The introduction of insulin with carbacetam significantly reduced the expression of VEGF and the development of specific morphological manifestations of DR. Thus, restoration of GABA-ergic signaling can be used as a promising therapeutic option for the correction of DR disorders.
miR-329-containing exosomes derived from breast tumor cells suppress VEGF and KDM1A expression in endothelial cells
N. Maleki1,2,3*, F. Karami1, S. Heyati2, M. HadiZadeh3, Gh. Parnian4*
1Department of Cellular and Molecular Biology, Faculty of Biological Sciences, Islamic Azad University-Tehran North Branch, Tehran, Iran;
*e-mail: dr.nargesmaleki@yahoo.com;
2Gynecology and reproductive biology Department, Kowsar poly-clinic, Tehran, Iran;
3Cancer Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran;
4Appletree Medical group, 275 Dundad W (Grange), Toronto, Ontario, Canada;
*e-mail: ghazalehparnian1@gmail.com
Received: 03 February 2021; Accepted: 07 July 2021
The exosomal transfer of miRNAs from tumor cells is considered to modulate VEGF expression and angiogenesis in endothelial cells. The aim of our investigation was to focus exclusively on the ability of specific exosomal miR329 to regulate angiogenesis within breast tumor. All experiments were done on MCF-7 and HUVEC cell lines. Exosomes were derived from MCF-7 cells both untreated and treated with tamoxifen that is an effecrive suppressor of hormone receptor-positive breast cancer. The level of miR32 and its targeted genes VEGF and lysine (K)-specific demethylase 1A (KDM1A) expression was estimated with q-RT-PCR. The PKH26 red fluorescent labeling kit was used to label the isolated exosomes and monitor their uptake. It was shown that the relative amount of miR-329 in exosomes was twice as large as in breast cancer cells. Fluorescence microscopy imaging presented that exosomes from MCF-7 cells were able to penetrate into endothelial cells and concentrate in the cytoplasm. It was observed that exosomes derived from untreated breast cancer cells induced KDM1A and VEGF gene expressions whereas exosomes from tamoxifen-treated cancer cells induced time-dependent decrease of KDM1A and VEGF expression in endothelial cells. It is assumed that the transfer of miR-329 containing exosomes from tamoxifen treated breast cancer cells to the endothelial cells could repress angiogenic molecular signaling pathway and be used as a supplementary strategy in breast cancer treatment.