Pathogenesis Of Varicose Veins Research Articles

Generation of a human induced pluripotent stem cell (iPSC, DVSi001-A) with a heterozygous mutation in KRAS (A209T)

Human varicose veins are commonly claimed to be responsible for lower limb symptoms. Mutation in KRAS gene has been implicated in various diseases, including cancers and vascular diseases. While little known about the novel mutation in KRAS gene and its contribution to the development of varicose veins. Here, we have generated human induced pluripotent stem cell (iPSC) line, which harboured a novel mutation in KRAS (c.209A>T) gene. This cell line provided a novel tool for understanding the mechanism of KRAS mutation in the pathogenesis of varicose veins.

Exploring causal correlations between inflammatory cytokines and varicose veins: A Mendelian randomization analysis.

This study aimed to investigate the causal relationship between inflammatory cytokines and the risk of varicose veins. The data were sourced from genome-wide association studies (GWAS) of European individuals. Multiple Mendelian randomization (MR) methods were used to evaluate the association between inflammatory cytokines and varicose veins. The study found significant associations between elevated levels of certain inflammatory biomarkers (e.g., CASP-8, Vascular endothelial growth factor A levels (VEGF_A)) and an increased risk of varicose veins, while others (e.g., 4EBP1, MMP-10) showed a protective effect. The MR-Egger Intercept and heterogeneity tests indicated no significant pleiotropy or heterogeneity. This comprehensive MR analysis identifies several cytokines as potential contributors to the pathogenesis of varicose veins, offering insights into novel therapeutic targets. Our findings underscore the importance of inflammation in varicose veins and suggest that targeting specific cytokines could be a promising strategy for the treatment and prevention of varicose veins.

Surgical Treatment of Varicose Veins of the Lower Extremities

The review presents an analysis of relevant literature on the surgical treatment of varicose veins of the lower extremities. Modern methods of surgical intervention for varicose veins are considered: open operations, methods of thermal ablation of the main veins, non-thermal non-tumescent methods, vein-preserving surgery (ASVAL), methods of treating reflux in perforating veins and recurrent varicose veins. The effectiveness of each of the surgical treatment methods used was analyzed. The frequency of relapses and the likelihood of complications of the described operations are considered. All surgical treatment methods presented in the review were developed on the basis of modern ideas about the pathogenesis of varicose veins, the mechanisms of formation of chronic venous insufficiency, and have an evidence base. These techniques are reflected in the latest clinical guidelines and are widely used in medical practice.

A novel view to varicose veins pathogenesis: Proteomic profiling suggests a pivotal role of extracellular matrix degradation.

Although morphological and anatomical studies indicate that venous wall weakening and subendothelial fibrosis characterize varicose veins (VV), the pathogenesis of VV remains poorly understood. The aim of this study is to obtain protein expression profiles in patients with VV and thereby get a step closer to understanding the pathogenesis of VV. Specimens were obtained from total of 10 patients, that is, from 5 patients undergoing VV surgical stripping and from 5 non-VV patients undergoing bypass surgery. Specimens were collected from the same layers of venous wall. Proteins were extracted from each specimen and analyzed by ion mobility spectrometry (IMS-MS). In total, 1387 were identified and 486 proteins were identified in all samples. From these, 15 proteins were differentially expressed between VV and non-VV samples (p < .05) and 12 of these showed a fold change >1.5. Interestingly, among the differentially expressed proteins, only two proteins were significantly increased in the VV tissue, that is, GAPDH (p = .028, fold change 2.74), where several proteins involved in maintaining the homeostasis in the extracellular matrix, that is, the CXXC zinc finger protein 5 (CXXC5) and nucleoporin (SEH1) were prominently downregulated (p = .049, fold change 37.8, and p = .040, fold change 3.46). The downregulation in protein expression of CXXC5 and SEH1 as well as upregulation of GAPDH were validated by Western blotting. The identified differentially expressed proteins suggest an altered profile of the connective tissue proteins as well as an increased proteolytic enzyme activity which both may be central in the pathophysiology of varicose veins.

Interleukin-13: association with inflammation and cysteine proteolysis in varicose transformation of the vascular wall

The present review considers current data on the structure, functions and role of interleukin-13 in the pathogenesis of vascular wall varicose transformation in terms of proteolysis and inflammatory response. It is known that interleukin-13 is able to interact with transforming growth factor-1 in diseases associated with fibrosis. The latter activates fibroblasts and excessive formation of the extracellular matrix, thereby inducing fibrosis of the vascular wall, which is one of the links in the pathogenesis of varicose veins. Also, to date, there is evidence of the interleukin-13 participation in the induction of certain proteolytic enzymes synthesis, such as matrix metalloproteinases. For the latter, participation in the transformation of the venous wall has been proven to date. The remodeling of the venous wall itself can lead to an increase in the expression of proteinases, providing a proteolytic mechanism for changing the structural organization of the venous wall in varicose veins of the lower extremities. At the same time, the involvement of lysosomal cysteine proteinases remains poorly understood. The expression and production of individual cathepsins are regulated by biologically active molecules: interleukin-1, interleukin-6, tumor necrosis factor , which are directly involved in inflammatory reactions in the wall of varicose veins. In particular, venous pathology develops in a vicious circle of inflammation with the formation of abnormal venous blood flow, chronic venous hypertension and dilation, and the recruitment of leukocytes. This leads to a further, deeper, remodeling of the walls and valves of the veins, an increase in blood pressure and the release of pro-inflammatory mediators chemokines and cytokines. In connection with the above, in order to understand the mechanisms of proteolysis in the vascular wall in varicose veins of the lower extremities, it is important to have an idea about the possible interactions of interleukin-13 with transforming growth factor-1, inflammatory cytokines, and cathepsins.

Роль эндотелиальной дисфункции и воспаления при хроническом заболевании вен нижних конечностей (обзор литературы)

INTRODUCTION: An analysis of publications of domestic and foreign literature was carried out and a review on endothelial dysfunction in patients with varicose saphenous veins of the lower extremities was presented. We searched Elibrary and PubMed biomedical publication databases for the required information for the last 5 to7 years using keywords: varicose saphenous veins of lower extremities, endothelial dysfunction, endothelial dysfunction markers. Today, diseases of the cardiovascular system occupy a leading position with the most common of them being chronic venous disease that affects 10% to 30% of the adult working-age population of the Russian Federation. Varicose veins cover a wide range of conditions from minor telangiectasia to pronounced varix dilatation of veins and severe venous insufficiency with venous ulcers. The endothelium is not only a barrier between blood and tissues, but also a kind of endocrine organ that produces a number of vasoactive substances that maintain homeostasis and perform, depending on environmental conditions, both an antithrombotic, anti-inflammatory, vasodilating effects, and a procoagulant, inflammatory and vasoconstrictive effects. Endothelial dysfunction is considered by many domestic and foreign authors as the main link in the development and progression of many diseases, including varicose veins. CONCLUSION: Determination of the dynamics of some markers of endothelial dysfunction and connective tissue dysplasia in patients with varicose saphenous veins of the lower extremities is a priority direction in prevention of disease progression. Despite ongoing research, many aspects of the pathogenesis of varicose veins of the lower extremities are still unclear. Further study of the basic factors of the development and progression of the pathology of the venous system is important for elaboration of algorithms for effective pathogenetic correction and improving the quality of life in patients with varicose veins of the lower extremities.

Monocyte chemoattractant protein 1 plasma concentration in blood from varicose veins decreases under venoactive drug treatment.

Vein-specific inflammation leads to vascular smooth muscle cells proliferation and extracellular matrix degradation of vein wall. This process is known as remodeling and is promoted by "trapped" leukocytes. Monocyte chemoattractant protein 1 (MCP-1) is a chemokine responsible for trafficking of leukocytes from blood to vein wall. The aim of this study was to measure the MCP-1 concentration in varicose veins blood before and after venoactive drug therapy and to compare it with a concentration of blood from varicose veins of subjects who did not receive drug treatment. Non-randomized comparative study was conducted on 30 patients with primary varicose veins. 20 patients of the study group received diosmin 900 mg/hesperidin 100 mg once daily. 10 controls received no treatment. MCP-1 level was measured (pg/mL) in the blood from varicose veins twice, at the day of inclusion and after 60 days. Legs discomfort related to chronic venous disease (CVD) symptoms was measured with 10-cm Visual Analogue Scale (VAS) at inclusion and at completion of the study. Median (interquartile range, IQR) MCP-1 concentrations in treatment and control groups at inclusion were 171.9 (124.4-216.0) and 157.0 (120.1-163.1), resp., P=0.285. After 60 days of treatment MCP-1 level decreased, but non-significantly to 152.3 (124.1-178.3). In patients who did not receive treatment chemokine level slightly increased to 163.0 (134.0-172.9). Median changes over time were -6.6 (-30.9-7.4) and 10.6 (-3.7-19.2) in the study and control groups, resp. (P=0.048). After 60 days in 12 of 19 and 2 of 9 patients of treatments and control groups MCP-1 decreased (P=0.103). Odds ratio for MCP-1 decreasing was 9.5 (95% CI 1.1-81.5, P=0.043) for those who received venoactive drug. Mean (± standard deviation [SD]) legs discomfort significantly dropped in the study group from 5.7 (±2.5) to 1.9 (±2.2) (P=0.0003), while in controls no changes were registered: 3.4 (±1.3) and 3.5 (± 1.4), resp., P=0.28). Mean difference of VAS at baseline and at follow-up was -3.5 (±2.6) and 0.9 (±2.1), resp. (P<0.0001). Plasma concentration of MCP-1 in varicose veins blood demonstrates a tendency to decrease under two months treatment with a venoactive drug. Future studies are needed to reveal a possible role of MCP-1 as a target considering its role in varicose veins pathogenesis.

Quantitative and structural characteristics of mitochondrial DNA in varicose veins

ObjectiveWe examined quantitative (in terms of mtDNA/nuclear DNA) and structural (in terms of common deletions in the MT-ND4 gene region) characteristics of mitochondrial DNA (mtDNA) in varicose veins (VVs) and venous wall layers by comparing mitochondrial genome parameters, as well as mitochondrial function (in terms of mitochondrial membrane potential (MtMP)), in varicose vein (VV) vs. non-varicose vein (NV) tissue samples. MethodsWe analyzed paired great saphenous vein samples (VV vs. NV segments from each patient left after venous surgery) harvested from patients with VVs. Relative mtDNA level and the proportion of no-deletion mtDNA were determined by a multiplex quantitative PCR (qPCR), confirming the latter with a more sensitive method – droplet digital PCR (ddPCR). Mitochondria's functional state in VVs was assessed using fluorescent (dependent on MtMP) live-staining of mitochondria in venous tissues. ResultsTotal mtDNA level was lower in VV than in NV samples (predominantly in the t. media layer). ddPCR analysis showed lower proportion of no-deletion mtDNA in VVs. Because of the decrease in relative MtMP in VVs, our results suggest a possible reduction of mitochondrial function in VVs. ConclusionQuantitative and structural changes (copy number and integrity) of mtDNA are plausibly involved in VV pathogenesis. Future clinical studies implementing the mitochondrial targeting may be eventually fostered after auxiliary mechanistic studies.

What is the relationship of varicose vein pathogenesis with collagen fibers?

In this study, the densities of collagen 1 and collagen 4, which are an effective vascular component in the remodelling of varicose veins, were investigated. The study included primary varicose vein samples of 20 patients and vein samples of 20 healthy controls. Immunohistochemical staining was performed using collagen 1 and collagen 4 antibodies. Histochemical staining was performed using Masson Trichrome. In the immunohistochemical analysis of varicose samples, collagen 1 immunostaining was negative in 17 cases (85%) and positive in 3 cases (15%). In healthy venous tissue samples, collagen 1 immunostaining was negative in 12 cases (60%) and positive in 8 cases (40%). There was no statistically significant difference between both groups concerning collagen 1 immunostaining (p > 0.05). In varicose samples, collagen 4 immunostaining was negative in 4 cases (20%) and positive in 16 cases (80%). In healthy venous tissue samples, collagen 4 immunostaining was negative in 13 cases (65%) and positive in 7 cases (35%). Statistical comparison of healthy veins and varicose veins concerning collagen 4 immunostaining showed a significant difference (p = 0.03). In the histochemical analysis of varicose samples, Masson Trichrome staining was negative in 4 cases (20%) and positive in 16 cases (80%). In healthy venous tissue samples, Masson Trichrome staining was negative in 18 cases (90%) and positive in 2 cases (10%). Statistical comparison of healthy veins and varicose veins concerning collagen 4 immunostaining showed a significant difference (p = 0.01). The change in the density of collagen types plays an important role in vein wall remodeling.

Signaling pathways associated with structural changes in varicose veins: a case-control study.

In varicose veins, blood pressure increases in the veins of the lower extremities due to mechanical stimulation and function remodeling. The aim of this study is assessment of Signaling pathways associated with structural changes in varicose veins. This pilot study was performed on patients with varicose veins, which had undergone surgery. The healthy tissues from trauma patients or vascular bypass without underlying diseases were used for control samples. Hematoxylin-eosin, trichrome, and elastin staining were used for histopathological examination. The levels of MDA (malondialdehyde), total thiol, SOD (Superoxide dismutase) and NO (nitric oxide) level were measured using Elisa kits to evaluate the oxidative stress level. Gene expression levels of MMP2, MMP9, FOXO3a, APOE and p53 genes were determined using Real-time PCR. This study showed, the vascular Vein wall changes are visible in vascular collagen staining. Although these changes are observed in the structure of vascular wall collagen fibers, the accumulation of collagen and elastin was increased in the walls of varicose veins compared to the control group. The amount of nitric oxide and thiol were increased in the varicose group (P < 0.0001). The expression of metalloproteinase2 gene associated with extracellular matrix change was increased in varicose veins. However, the amount of metalloproteinase 9 was decreased in this group compared to control group. FOXO3a, APOE Genes were down-regulated in the varicose veins compared to control group, while p53 gene expression was significantly increased in the varicose group (P < 0.0001). This study demonstrated changes in oxidative stress, morphological structure, and aging pathways in varicose when compared to non-varicose veins. The changes in oxidative stress may be associated with the variations in morphological structure and aging pathways which contribute to the pathogenesis of varicose veins.

MicroRNA‑199a‑5p regulates FOXC2 to control human vascular smooth muscle cell phenotypic switch.

Varicose veins are among the most common disorders of the vascular system; however, the pathogenesis of varicose veins remains unclear. The present study aimed to investigate the roles of microRNA (miR)-199a-5p in varicose veins and in the phenotypic transition of vascular smooth muscle cells (VSMCs). Bioinformatics analysis confirmed that miR-199a-5p had target sites on the forkhead box C2 (FOXC2) 3′-untranslated region. Reverse transcription-quantitative PCR (RT-qPCR) and western blotting were used to detect the expression levels of miR-199a-5p and FOXC2 in varicose vein and normal great saphenous vein tissues. Cell Counting Kit-8 and Transwell migration assays were performed to validate the effects of miR-199a-5p on VSMCs. Contractile markers, such as smooth muscle 22α, calponin, smooth muscle actin and myosin heavy chain 11 were used to detect phenotypic transition. RT-qPCR revealed that miR-199a-5p was downregulated in varicose veins compared with expression in normal great saphenous veins, whereas FOXC2 was upregulated in varicose veins. In addition, biomarkers of the VSMC contractile phenotype were downregulated in varicose veins. Overexpression of miR-199a-5p by mimics suppressed VSMC proliferation and migration, whereas depletion of miR-199a-5p enhanced VSMC proliferation and migration. Notably, the effects caused by miR-199a-5p could be reversed by FOXC2 overexpression. Dual luciferase reporter analysis confirmed that FOXC2 was a target of miR-199a-5p. In conclusion, miR-199a-5p may be a novel regulator of phenotypic switching in VSMCs by targeting FOXC2 during varicose vein formation.

Transforming growth factor-β1 and inducible nitric oxide synthase signaling were involved in effects of prostaglandin E2 on progression of lower limb varicose veins

ObjectiveThe vital pathogenesis of varicose veins includes remodeling of the extracellular matrix and decreased vascular tone. Prostaglandin E2 (PGE2), a small molecule substance and inflammatory medium that belongs to the arachidonic acid derivatives, has the capacity to influence the expression of metalloproteinase and the vascular tone of the venous wall. The purpose of the present study was to investigate the role of PGE2 in the development of varicose veins in lower limbs. MethodsThe collected venous specimens were analyzed using hematoxylin and eosin, Masson's trichrome, and immunohistochemical staining. Transforming growth factor (TGF)-β1, PGE2, CD31, and α-smooth muscle actin antibody were used to detect the expression and distribution of these proteins. The effect of PGE2 on the proliferation, migration, and tube formation capacity of human umbilical vein endothelial cells (HUVECs) was detected in vitro. The effect of TGF-β1 on the expression of PGE2 and matrix metalloproteinases (MMPs) was assessed using Western blotting. Quantitative reverse transcription polymerase chain reaction was used to evaluate the effect of PGE2 on the expression of nitric oxide synthase (NOS) and other genes. ResultsThe expression of PGE2 and TGF-β1 in varicose veins was upregulated in the media tunica and intima tunica, and a strong positive correlation was found between PGE2 and TGF-β1 expression in both varicose veins (95% confidence interval, 0.5207-0.9582; R = 0.848; P = .0005) and normal veins (95% confidence interval, 0.2530-0.8532; R = 0.643; P = .003). PGE2 promoted the migration and tube formation ability of HUVECs. Moreover, PGE2 also upregulated the expression of MMP-1 and TGF-β1 in HUVECs and increased the mRNA level of inducible NOS. ConclusionsPGE2 can affect the remodeling of the extracellular matrix and reduce the elasticity of the vascular walls by promoting the synthesis of TGF-β1 and MMP-1. PGE2 can also reduce the tension of the great saphenous vein by promoting the expression of inducible NOS, thus aggravating the blood stasis.

Cytokines in Pathogenesis of Varicose Veins

<h3></h3> Патогенез варикозной болезни до сих пор остается до конца не изученным и служит предметом активного исследования клиницистов и ученых фундаментальных направлений медицины. Причиной узловатой трансформации признают ремоделирование венозной стенки, которое, в свою очередь, развивается в результате так называемого веноспецифического воспаления. Этот хронический процесс включает комплекс взаимодействий между клетками, отвечающими за воспалительную реакцию (нейтрофилы, лимфоциты, моноциты, макрофаги), эндотелиальными клетками, гладкомышечными клетками сосудов и внеклеточным матриксом. Кроме того, веноспецифическое воспаление протекает при активном участии цитокинов, которые представляют собой разнообразную группу небольших белков, продуцируемых разными иммунными и сосудистыми клетками, которые, действуя в пикомолярных и наномолярных концентрациях, активируют специфические рецепторы и модулируют функции многих клеток и тканей. Роль цитокинов в патогенезе варикозной болезни остается малоизученной ввиду сложности изучения их механизма действия. Предполагается, что их активность опосредует нарушения регулирования деградации внеклеточного матрикса, видоизменение составляющих внеклеточного матрикса, таких как коллаген и эластин, снижение количества гладкомышечных клеток, что приводит к снижению эффективности сокращения сосудистой стенки из-за фрагментации мышечного слоя вены. Цитокины участвуют и в процессах активации лейкоцитов, что приводит к высвобождению свободных радикалов, активации протеазы и последующей деградации гладкомышечных клеток. В рамках данной работы представлена общая информация о цитокинах, а также проведен анализ исследований, в которых изучено возможное влияние различных молекул на развитие первичного варикозного расширения вен.

The genetic constituent of varicose vein pathogenesis as a key for future treatment option development

Vessel Plus is an open acccess journal, which publishes articles related to vascular diseases, including acute respiratory distress syndrome, aneurysm, atherosclerosis, hypertension, stroke, peripheral vascular or pulmonary vascular diseases, etc.

Sex-Specific Genetically Predicted Iron Status in relation to 12 Vascular Diseases: A Mendelian Randomization Study in the UK Biobank.

Background Iron overload has been implicated in the pathogenesis of varicose veins (VVs). However, the association of serum iron status with other vascular diseases (VDs) is not well understood, which might be a potential target for VD prevention. This study was aimed at investigating the causal associations between iron status and VDs using the Mendelian randomization (MR) method. Methods A two-sample MR was designed to investigate whether iron status was associated with VDs, based on iron data from a published genome-wide association study meta-analysis of 48,972 subjects of European descent and VD data obtained from the UK Biobank, including 361,194 British subjects (167,020 males and 194,174 females). We further explored whether there was sex difference in the associations between genetically predicted iron status and VDs. Results The results demonstrated that iron status had a significant causal effect on VVs of lower extremities (P < 0.001) and a potential effect on coronary atherosclerosis (P < 0.05 for serum iron, ferritin, and transferrin saturation, respectively), but not on other VDs. Furthermore, higher iron status exerted a detrimental effect on VVs of lower extremities in both genders (P < 0.05) and a protective effect on male patients with coronary atherosclerosis (P < 0.05 for serum iron, ferritin, and transferrin saturation, respectively). Conclusions This MR study provides robust evidence that higher iron status increases the risk of VVs of lower extremities, whereas it reduces the incidence of coronary atherosclerosis in the male population, which indicates that iron has divergent effects on vascular pathology.

Altered Expression of the Extracellular Matrix Related Genes COL15A1, CHRDL2, EFEMP1, and TIMP1 in Varicose Veins

Background. Molecular mechanisms underlying pathogenesis of varicose veins are not fully understood. Expression of genes in varicose veins compared with normal vein segments in patients with varicose veins was evaluated. The pilot study using micro-array analysis revealed a set of genes differentially expressed in varicose veins. In the present study The aim was to validate micro-array data on genes related to extracellular matrix: COL15A1 (collagen type XV alpha 1 chain), CHRDL2 (chordin like 2), EFEMP1 (EGF containing fibulin extracellular matrix protein 1), and TIMP1 (metallopeptidase inhibitor 1), using an independent method and a replication set. Methods. Paired GSV samples (n = 26, varicose and non-varicose segments) harvested from 13 patients with varicose veins (C2) were used in this study. Patients with post-thrombotic changes in deep veins and absence of visible varicose veins were excluded. Determination of the level of mRNA expression (normalised to ACTB and GAPDH) was performed by reverse transcription real time qPCR. Statistical analysis was performed with Wilcoxon signed rank test using the qBase+ software. The work was supported by the Russian Science Foundation (Project 17-75-20223 “Investigation of the mechanisms of vein wall remodelling in varicose veins”). Results. Significantly higher mRNA levels of COL15A1 (1.46 fold, CI 1.21–1.76), CHRDL2 (2.15 fold, CI 1.37-3.39), EFEMP1 (2.01 fold, CI 1.29–3.14), and TIMP1 (1.58 fold, CI 1.15–2.17), (p < .005) were observed. These findings are consistent with previous transcriptome analysis. Conclusion. The extracellular matrix related genes COL15A1, CHRDL2, EFEMP1, and TIMP1 are up-regulated in varicose veins which plausibly contributes to the pathogenesis.

IGFBP6 regulates vascular smooth muscle cell proliferation and morphology via cyclin E-CDK2.

Despite the high prevalence of varicose veins, the underlying pathogenesis of this disease remains unclear. The present study aims to explore the role of insulin-like growth factor binding protein 6 (IGFBP6) in vascular smooth muscle cells (VSMCs). Using a protein array approach, we identified several differentially expressed proteins between varicose great saphenous veins and normal great saphenous veins. Bioinformatic analysis showed that IGFBP6 was closely related to cell proliferation. Further validation confirmed that IGFBP6 was one of the most highly expressed proteins in varicose vein tissue. Knocking down IGFBP6 in VSMCs significantly attenuated cell proliferation and induced the S phase arrest during the cell cycle. Further experiments demonstrated that IGFBP6 knockdown increased cyclin E ubiquitination, which reduced expression of cyclin E and phosphorylation of CDK2. Furthermore, IGFBP6 knockdown arrested centrosome replication, which subsequently influenced VSMC morphology. Ultimately, IGFBP6 was validated to be involved in VSMC proliferation in varicose vein tissues. The present study reveals that IGFBP6 is closely correlated with VSMC biological function and provides unprecedented insights into the underlying pathogenesis of varicose veins.

Elevated c-fos expression is correlated with phenotypic switching of human vascular smooth muscle cells derived from lower limb venous varicosities

BackgroundLower limb venous varicosities (VVs) are clinically common; however, their molecular underpinnings are far from well elucidated. Previous studies have demonstrated that the phenotypic transition of vascular smooth muscle cells (VSMCs) plays a critical role in VV pathogenesis and that c-fos is upregulated in VSMCs from VVs. The present study investigated the histologic and cytologic changes in VVs and the correlation between c-fos upregulation and VSMC phenotypic switching. MethodsThirty-four patients with VVs (VV group) and 13 patients undergoing coronary artery bypass using autologous great saphenous vein segments (normal vein [NV] group) were enrolled in the present study. The great saphenous veins of both groups were harvested for subsequent experiments. Hematoxylin and eosin staining was performed for vein morphologic analysis. Real-time quantitative polymerase chain reaction, immunohistochemistry, and Western blot assays were used to assess mRNA and protein expression of c-fos, α-smooth muscle actin (α-SMA), and osteopontin (OPN). Simple linear regression was used to evaluate the correlation between c-fos and OPN/α-SMA. Primary VSMCs were isolated from both groups and cultured in vitro. A cell counting kit-8 assay and scratch-wound assay were used to analyze the proliferation and migration abilities of the cells, respectively. ResultsThe mean age of the patients in the NV and VV groups was 61.4 ± 3.8 years and 59.5 ± 10.4 years, respectively. The vein cavities of the VV group were dilated, and the arrangement of the cells was disordered. The tunica media of the VV group was thicker than that of the NV group owing to the accumulation and proliferation of VSMCs. Significantly elevated mRNA levels of c-fos and OPN were observed in the VV group compared with the NV group, and a positive correlation was further demonstrated between the mRNA levels of c-fos and OPN/α-SMA (R2, 0.5524; P < .001). The VSMCs derived from the VV group were more numerous (as shown by the cell counting kit-8 assay) and had a significantly greater migration speed (as shown by the scratch-wound assay) than those derived from the NV group. Moreover, the protein expression of c-fos was significantly upregulated in VSMCs derived from the VV group, and this change was accompanied by a decrease in α-SMA and an increase in OPN expression. ConclusionsBoth mRNA and protein expression of c-fos were upregulated in VV specimens, and the phenotypic biomarkers (OPN/α-SMA) were altered concurrently. VSMCs derived from VVs showed increased proliferation and migration abilities. Upregulation of c-fos might play a role in the phenotypic switching of VSMCs and subsequently participate in the pathogenesis of VVs. Clinical RelevanceC-fos is an immediate early gene owing to the transient and rapid change in its expression in response to stimuli. It is involved in the regulation of cell proliferation, cell growth, and cell movement. In the present study, varicose vein specimens showed increased mRNA and protein expression of c-fos, accompanied by altered phenotypic biomarkers. The upregulation of the c-fos gene in smooth muscle cells cultured from varicose vein specimens might be associated with phenotypic switching and functional disturbance. These results could contribute to the exploration of the molecular mechanisms underlying the pathogenesis of varicose veins and the development of new therapeutic strategies.

Varicose veins: on the verge of discovering the cause?

Varicose veins of the lower limbs are one of the most common and wide-spread pathology all around the world. What triggers the specific changes in a vein wall still remains unclear as well as what happens in the layers of the vein wall after the disease starts. The aim of the article is to analyze published data and results of researches on epidemiology, genetics, cellular and molecular mechanisms underlying varicose veins pathogenesis. It is now commonly accepted that vein wall changes in patients with varicose veins result from vein-specific inflammation. This process includes leukocytes adhesion to venous endothelium with their subsequent migration into the vein wall and surrounding tissues. Activated leukocytes express a number of molecules that lead to vein wall remodeling and dilation. Comprehensive assessment of the epidemiological data on the prevalence of varicose veins and risk factors, of the findings from genetic studies, of data on molecular-cell interactions as well as results of various surgical interventions in patients with varicose veins, shows that remodeling is a reversible process that can be stopped and reversed by different stimuli including some chemical substances. For the first time in the literature, the authors assume that varicose veins can be successfully cured pharmacologically with no surgical interventions needed.

Histological, Immunohistochemical, Morphometric and Molecular Changes in the Wall of Human Varicose Veins

Abstract Background: Varicose veins (VVs) are a major chronic venous disease, yet the underlying mechanisms are unclear. This disorder is characterized by progressive architectural remodeling of the vessel wall. Recent studies have focused on the role of matrix metalloproteinases (MMPs) in the pathogenesis of VVs. Aim of Study: The present study was assigned to demon-strate the morphologic and morphometric changes in the wall of VVs and to correlate these changes with the expression of MMP-1 and MMP-9. Material and Methods: A total number of 18 human long saphenous vein samples (8 normal and 10 VVs) were included in the study. The specimens were processed and examined histologically using hematoxylin and eosin, Masson's trichrome and orcein, together with immunohistochemistry for alpha smooth muscle actin (Eo-SMA). The mRNA expression of MMP-1 and MMP-9 was analyzed using real-time polymerase chain reaction (PCR). Quantitative morphometric measure-ments were carried out by the image analyzer. Results: The wall of VVs showed damaged endothelial surface, interrupted internal elastic lamina and numerous vasa vasora. There was significant increase in intima and media thickness compared to normal veins. Significant increase in the mean collagen area percent and decrease in elastin area percent was also observed. The mean area percent of Eo-SMA immunostaining and count of smooth muscle cells (SMCs) showed significant increase, indicating SMCs hypertrophy and hyperplasia, while the optical density of Eo-SMA immu-nostaining decreased. These changes were associated with significant increase in mRNA expression of MMP-1 and MMP-9. Conclusion: Varicose veins showed marked disorganiza-tion in all components of the vessel wall (collagen, elastin and SMCs). The associated increase in mRNA expression of MMPs might be one of the molecular mechanisms underlying vascular remodeling.