Vascular Endothelial Lesions Research Articles

Anti-atherosclerotic effect of alfalfa flavonoid extract by regulating inflammation and oxidative stress in HUVEC cells and rats

Atherosclerosis is characterized by accumulation of lipid that triggers arterial inflammation. This study aims to investigate the protective effect of alfalfa flavonoid extract (AFE) on HUVEC cells treated with lipopolysaccharide (LPS) and rats fed with high-fat diet (HFD). AFE reduced the expression of proinflammatory factors (COX-2, iNOS, IL-1β, IL-6 and TNF-α), intercellular adhesion molecule-1 and vascular cell adhesion molecule-1, and inhibited activation of NF-κB and MAPK pathways. AFE alleviated vascular endothelial lesions by reducing HFD-enhanced levels of triglyceride, total cholesterol and low-density lipoprotein but increased high-density lipoprotein. Mechanically, AFE ameliorated HFD-enhanced liver steatosis through downregulating gene expression for lipid biosynthesis (SREBP2, HMGCR and FASN) and lipid uptake (LDLR), and through upregulating CYP7A1 expression for lipid catabolism and excretion. These data suggest that AFE possesses the anti-atherosclerotic potential through controlling inflammation, hyperlipidemia and lipid metabolism, and can be used as a functional food additive to benefit patients with atherosclerosis.

Helicobacter pylori components increase the severity of metabolic syndrome and its hepatic manifestations induced by a high fat diet

The metabolic syndrome, often accompanied by hepatic manifestations, is a high-risk factor for developing cardiovascular disease. Patients with metabolic dysfunction associated with steatohepatic disease (MASDL) are at significant risk of developing coronary artery disease. Atherosclerosis is a systemic inflammatory disorder in which several factors, including dietary or infectious factors, can cause an inflammatory response. Helicobacter pylori (HP) bacteria have been implicated in the progression of proatherogenic vascular endothelial lesions, moreover, our previous study in an experimental in vivo model of Cavia porcellus showed that HP components and high-fat substances acted synergistically in promoting vascular endothelial inflammation, leading to an early onset of a proatherogenic environment. In the present study, our goal was to determine the contribution of HP components to the development of hepatic manifestations of metabolic syndrome in an experimental model. Our results showed that HP infection in animals exposed to a high-fat diet increased oxidative stress and lipid peroxidation, followed by endothelial lipid deposition, impaired endothelial apoptosis, cell lysis, and increased vascular stiffness. Finally, histopathological analysis of liver tissue showed signs of MASLD development in HP-infected animals fed a high-fat diet.

Sodium Selenite Ameliorates Silver Nanoparticles Induced Vascular Endothelial Cytotoxic Injury by Antioxidative Properties and Suppressing Inflammation Through Activating the Nrf2 Signaling Pathway

Silver nanoparticles (AgNP) are the dominant nanomaterials in commercial products and the medical field, but the widespread occurrence of AgNP has become a global threat to human health. Growing studies indicate that AgNP exposure can induce vascular endothelial toxicity by excessive oxidative stress and inflammation, which is closely related to cardiovascular disease (CVD), but the potential intrinsic mechanism remains poorly elucidated. Thus, it has been crucial to control the toxicological effects of AgNP in order to improve their safety and increase the outcome of their applications.Multiple researches have demonstrated that sodium selenite (Se) possesses the capability to counteract the toxicity of AgNP, but the functional role of Se in AgNP-induced CVD is largely unexplored. The aim of this study was to explore the potential protective effect of Se on AgNP-induced vascular endothelial lesion and elucidate the underlying mechanisms. An in vivo model of toxicity in animals was established by the instillation of 200 µL of AgNP into the trachea of rats both with (0.2 mg/kg/day) and without Se treated. In vitro experiments, human umbilical vein endothelial cells (HUVECs) were incubated with AgNP (0.3 µg/mL ) and Se for a duration of 24 h. Utilizing transmission electron microscopy, we observed that the internalization of AgNP-induced endothelial cells was desquamated from the internal elastic lamina, the endoplasmic reticulum was dilated, and the medullary vesicle formed. Se treatment reduced the levels of vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1), inhibited the release of pro-inflammatory cytokines (specifically tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6), improved endothelial cell permeability, integrity, and dysfunction, and prevented damage to the aortic endothelium caused by AgNP. Importantly, we found that Se showed the capacity against AgNP with biological functions in guiding the intracellular reactive oxygen species (ROS) scavenging and meanwhile exhibiting anti-inflammation effects. Se supplementation decreased the intracellular ROS release and suppressed NOD-like receptor protein 3 (NLRP3) and nuclear factor kappa-B (NF-κB) mediated inflammation within AgNP-intoxicated rats and HUVECs. The anti-oxidant stress and anti-inflammatory effects of Se were at least partly dependent on nuclear factor erythroid 2-related factor 2 (Nrf2). Overall, our results indicated that the protectiveness of Se against AgNP-induced vascular endothelial toxicity injury was at least attributed to the inhibition of oxidative ROS and pro-inflammatory NF-κB/NLRP3 inflammasome by activating the Nrf2 and antioxidant enzyme (HO-1) signal pathway.

Physiological and pathological characteristics of vascular endothelial injury in diabetes and the regulatory mechanism of autophagy.

Vascular endothelial injury in diabetes mellitus (DM) is the major cause of vascular disease, which is closely related to the occurrence and development of a series of vascular complications and has a serious negative impact on a patient's health and quality of life. The primary function of normal vascular endothelium is to function as a barrier function. However, in the presence of DM, glucose and lipid metabolism disorders, insulin resistance, inflammatory reactions, oxidative stress, and other factors cause vascular endothelial injury, leading to vascular endothelial lesions from morphology to function. Recently, numerous studies have found that autophagy plays a vital role in regulating the progression of vascular endothelial injury. Therefore, this article compares the morphology and function of normal and diabetic vascular endothelium and focuses on the current regulatory mechanisms and the important role of autophagy in diabetic vascular endothelial injury caused by different signal pathways. We aim to provide some references for future research on the mechanism of vascular endothelial injury in DM, investigate autophagy's protective or injurious effect, and study potential drugs using autophagy as a target.

1,8-Cineole ameliorates endothelial injury and hypertension induced by L-NAME through regulation of autophagy via PI3K/mTOR signaling pathway

Our previous data confirmed that 1,8-Cineole had an antihypertensive effect in animal models. However, it is unclear whether antihypertension is dependent on the protective effect of 1,8-Cineole on endothelial function and structure. At present, the purpose was to investigate the protective effects of 1,8-Cineole on vascular endothelial tissue in hypertensive rats and human umbilical vein endothelial cells (HUVECs). Our results showed that 1,8-Cineole significantly reduced the blood pressure and improved the vascular endothelial lesion, attenuated vascular oxidative stress and inflammation induced by Nω-Nitro-L-arginine methyl ester hydrochloride (L-NAME) in rats. Pretreatment with 1,8-Cineole was able to inhibit the increase in malondialdehyde (MDA) and reactive oxygen species (ROS) induced by L-NAME, and increased the release and expression of superoxide dismutase (SOD) and nitric oxide (NO). In addition, 1,8-Cineole also reversed the increase of autophagy-associated protein LC3Ⅱ/LC3Ⅰ and the decrease of P62 in vivo and in vitro respectively. There was a synergistic effect between PI3K agonists and drugs, while PI3K inhibitors blocked the efficacy of 1,8-Cineole. The addition of autophagy inhibitor chloroquine increases the expression of eNOS. Taken together, our results indicate that 1,8-Cineole has potential beneficial promising antihypertension depending on the integrity of vascular endothelial structure and function induced by L-NAME, and the mechanism involves ameliorating autophagy by regulating of PI3K/mTOR.

Role of profilin-1 in vasculopathy induced by advanced glycation end products (AGEs)

AimsTo construct a simple and feasible rat model to mimic diabetic vasculopathy by chronic injection of advanced glycation end products (AGEs) and further determine the role of profilin-1 in vasculopathy in AGE-injection rats. MethodsSprague-Dawley rats were injected with AGEs-BSA (25 mg/kg/day) for 0, 20, 30, 40, and 60 days by caudal vein. Then, the morphological changes in the aorta, heart, and kidney and the expression of profilin-1 were assessed. In cultured endothelial cells, shRNA profilin-1 was used to clarify the role of profilin-1 in AGEs-induced vascular endothelial lesions and inflammatory reactions. ResultsThe aorta, heart, and kidney of the AGE-injection rats had obvious morphological changes. Also, the indicators of vascular remodeling in the aorta significantly increased, accompanied by the increased expression of profilin-1 in the aorta, heart, and kidney and polysaccharide content on the kidney basement membrane. In addition, the protein level of profilin-1 was markedly upregulated in the aorta of AGEs-injected rats and endothelial cells incubated with AGEs. shRNA profilin-1 markedly attenuated the upregulated expression of profilin-1, receptor for AGEs (RAGE), and NF-κB in endothelial cells incubated with AGEs, as well as reduced the high levels of ICAM-1, IL-8, TNF-α, ROS, and apoptosis induced by AGEs. ConclusionsExogenous AGEs can mimic diabetic vasculopathy in vivo to some extent and increase profilin-1 expression in the target organs of diabetic complications. Blockade of profilin-1 attenuates vascular lesions and inflammatory reactions, suggesting its critical role in the metabolic memory mediated by AGEs.

Impact of endodontic and periodontal diseases and treatments on the aorta and liver of obese and non-obese rats.

To evaluate the impact of the presence and treatment of periodontal disease (PD) and apical periodontitis (AP) on the aorta and liver of obese and non-obese rats. One hundred and forty Wistar rats were divided into two groups, according to the diet administered: normal diet (-n), without obesity; and cafeteria diet (-c), with induced obesity. These groups were divided into seven subgroups according to the specific experimental protocols: naïve control (NC); AP; AP with treatment (APt); PD; PE with treatment (PDt); AP and PD (APPD); and AP and PD with treatment (APPDt). AP and PD lesions were induced for four weeks. Four weeks after treatments, the animals were euthanatized, and the aorta and liver were dissected for histological evaluation. For the comparison of the thickness of the aorta between groups, the Kruskal-Wallis test was used, followed by the Mann-Whitney test. For the analysis of other variables related to the aorta and liver outcomes, logistic regression was carried out. Both PD and AP were associated with the development of histological alterations in the aortic arch, with no significant difference between obese and non-obese animals (p=.17). The aorta thickness was increased significantly (p<.05) with the combination of PD and AP in obese rats (APPDt-c group) compared with the other groups (NC-n, APt-n, APt-c and AP-c). The logistic regression models revealed that the untreated (OR=7.78; 95%CI=2.4-25) and treated (OR=2.9; 95%CI=1.0-8.4) groups were significantly more likely to have endothelial alterations compared with the control groups (p=.002). Obesity (OR=16.5; 95%CI=3.4-81.3) was the only predictor variable of liver steatosis (p<.001). Histological alterations in the aortic arch of obese and non-obese rats were observed in the presence of periodontal disease and apical periodontitis. The combination of PD and AP increased the aorta thickness in obese rats. A reduction of vascular endothelial lesions was observed with the treatments of PD and AP.

Survivin regulated by autophagy mediates hyperglycemia-induced vascular endothelial cell dysfunction

Diabetic vascular complications are often defined by vascular endothelial lesions. However, as a plastic cell type, whether endothelial cells could transit from quiescence to hyper-active status and hamper vascular stability upon hyperglycemia stimulation and whether this process is involved in diabetic vascular complications remain obscure. Survivin has been identified as an anti-apoptotic protein in tumor or epithelial cells by either promoting proliferation or inhibiting apoptosis. Therefore, this study aims at investigating the effects of hyperglycemia on endothelial cell status and the potential involvement of survivin. We found that high glucose (25 mM) did not cause endothelial injuries, instead, it evidently promotes endothelial proliferation and tube formation capacity indicating endothelial cell dysfunction upon hyperglycemia characterized by its preference to hyper-active status. Concomitantly, an upregulation of survivin was detected accompanied by the key component elevations of autophagy pathway including LC3, Beclin1, and p62. YM155, a specific inhibitor of survivin, could abrogate hyperglycemia-induced endothelial hyper-activation. Application of the autophagy inhibitor (3MA) and agonist (rapamycin) supported that survivin could be as a downstream effect or of autophagy. Thus, our results suggested that survivin/autophagy axis a potential therapeutic target in treatment of diabetic vascular complications.

The effect of lysophosphatidic acid on Toll-like receptor 4 expression and the nuclear factor-κB signaling pathway in THP-1 cells.

Toll-like receptors (TLRs) are major receptors that mediate the innate immune and inflammatory responses, of which TLR4 has been found most closely related to human atherosclerosis. After ligands are polymerized and activated by TLR, the mitogen-activated protein kinase and nuclear factor-κB (NF-κB) pathways are activated, leading to promotion of NF-κB-regulated transcription of inflammatory factors, thus playing a role in the physiological and pathological processes in atherosclerosis. Oxidized lipoproteins or their components, oxidized lipids, have been confirmed as endogenous TLR receptors. Lysophosphatidic acid (LPA) is an active component of low-density lipoprotein that induces vascular endothelial lesions. However, the mechanism of the TLR4/NF-κB signaling system involved in LPA-induced atherosclerosis has not been fully elucidated. In this study, we investigated the effects of LPA on TLR4 expression, nuclear translocation of NF-κB p65 subunit, and changes in the cytokine tumor necrosis factor α (TNF-α) in human THP-1 cells. LPA upregulated expression of the TLR4 mRNA and protein in THP-1 cells in a dose- and time-dependent manner, induced NF-κB p65 activation synchronously in THP-1 cells, and increased TNF-α secretion. After TLR4 was blocked using TLR4 monoclonal antibody, NF-κB p65 expression and TNF-α secretion were inhibited significantly. These data suggest that LPA can significantly upregulate TLR4 expression and promote NF-κB activation and proinflammatory cytokine secretion in THP-1 cells; it is possible that the TLR4/NF-κB signaling pathway mediates the atherogenic effect of LPA.

Intravascular Papillary Endothelial Hyperplasia (Masson's Hemangioma) Presenting as Soft Tissue Mass in Left Scapular Region-An Unusual Presentation

Intravascular papillary endothelial hyperplasia (IPEH) or reactive vascular endothelial lesion is a rare benign condition and is also known as Masson’s hemangioma. It may appear as a primary or pure form developing in a distended vessel, or it can be associated with hemangiomas, pyogenic granulomas, or lymphagiomas. It is usually confined to the lumen of preexisting vessels or vascular malformations. Nearly all the lesions are intimately associated with a thrombus in various stages of organization. The main significance of IPEH is its clinical and histological resemblance to soft-tissue sarcoma and possible misinterpretation as such. We hereby report a case of IPEH clinically and radiologically misdiagnosed as soft tissue sarcoma, in a 47-year-old man, presenting as a soft tissue mass in the left scapular region.

Histopathological studies of microtubule disassembling agent-induced myocardial lesions in rats

Microtubule disassembling agents (MDAs) such as colchicine (COL) and vincristine sulfate (VCR) are known to be cardiotoxic. However, few attempts have been made to histopathologically examine cardiac lesions induced by MDAs. In this study, we endeavored to induce myocardial injury in rats by administering MDAs and to clarify the morphological features of these myocardial lesions. Male rats were intravenously administered COL (1.00 or 1.25mg/kg for 2 days at single daily doses) or VCR (0.50 or 0.75 mg/kg for 2 days at single daily doses). The day after administration, hearts were excised and examined histopathologically, immunohistochemically and electron microscopically. Degeneration and necrosis of myocardial cells with vacuolation were observed in rats administered COL at 1.25mg/kg or VCR at 0.75 mg/kg. Electron microscopic examination revealed vacuoles in swollen mitochondria. Moreover, there were cells showing pyknosis and karyorrhexis in the interstitium. TUNEL and immunohistochemical staining for endothelial cells and electron microscopic examination identified the apoptotic cells in the interstitium to be vascular endothelial cells. These vascular endothelial lesions were induced by lower doses of MDAs than were myocardial lesions. Furthermore, common sites of cardiac lesions induced by MDAs had almost the same distribution as areas positive for pimonidazole, a marker of hypoxia. These findings indicate that MDAs occasionally damage mitochondria in myocardial cells, and suggest that these changes involve microcirculatory dysfunction induced by endothelial cell injury.

Effects of Liandou Qingmai Recipe ( ) on Endothelin-1, Nitric Oxide, Interleukin-6 and Interleukin-10 Levels in Patients with Coronary Heart Disease

To observe effects of Liandou Qingmai Recipe (see text) on endothelin-1 (ET-1), nitric oxide (NO), interleukin-6 (IL-6) and IL-10 levels in patients with coronary heart disease. Total 101 cases with coronary heart disease were randomly divided into a treatment group (n = 45) treated by Liandou Qingmai Recipe and a standard treatment group (control group, n = 56), with a normal group of 16 health persons set up. Changes of ET-1, NO, IL-6 and IL-10 levels were measured before treatment and after treatment for two weeks. And the data were analyzed by SPSS 16.0 statistic software. Before treatment, the levels of ET-1, IL-6 and IL-10 levels were significantly higher and NO was significantly lower in the patients with coronary heart disease than those in the normal group (90.7 +/- 12.7 ng/L vs 41.8 +/- 13.5 ng/L, 9.17 +/- 0.18 ng/L vs 1.10 +/- 0.08 ng/L, 1.94 +/- 0.26 ng/L vs 1.09 +/- 0.06 ng/L, and 92.2 +/- 17.7 micromol/L vs 124.5 +/- 27.2 micromol/L; all P < 0.05), with no significant differences in the levels of ET-1, NO, IL-6 and IL-10 between the treatment group and the control group (P > 0.05); After treatment, ET-1 and IL-6 significantly decreased in the treatment group and the control group, and NO increased in the treatment group; And IL-6 level was significantly lower and NO level was higher in the treatment group than those in the control group (4.48 +/- 1.22 ng/L vs 5.13 +/- 1.85 ng/L, 117.4 +/- 22.3 micromol/L vs 92.4 +/- 17.1 micromol/L; both P < 0.05); There was a positive correlation between IL-6 and IL-10, and a negative correlation between NO and IL-10 (r = 0.142, r = -0.152; both P < 0.05). Liandou Qingmai Recipe can decline IL-6, IL-10 and ET-1 levels, and raise NO level in patients with coronary heart disease on the basis of standard treatment, so as to inhibit endothelial inflammatory response, improve vascular endothelial function, with stronger anti-AS action; And vascular endothelial lesion is related with inflammatory response.

Interaction between COX-2 and iNOS aggravates vascular lesion and antagonistic effect of ginsenoside

Aim of the study Ginseng, the root of Panax ginseng C.A.Meyer (Araliaceae), is one of the most widely used Chinese herbs with hypotensive and cardiotonic actions for thousands of years, but the underlying mechanisms have not been well determined. Ginsenoside, the effective components of ginseng, has anti-inflammatory and anti-oxidative effects. Cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) are key enzymes in inflammation and oxidative stress, respectively, which have close interaction, aggravating their damaging effects. This study investigated whether COX-2 interacted with iNOS in vascular endothelial lesion and the protective effect of ginsenoside. Materials and methods SD male rats were fed with high l-methionine (3%, w/w) to induce vascular endothelial lesion, and the rats in ginsenoside group were fed ginsenoside solution (0.8 mg kg −1 d −1). The mRNA expression and protein contents of COX-2 and iNOS were detected by RT-PCR and Western blotting, respectively. The interaction between COX-2 and iNOS was analyzed by co-immunoprecipitation and laser confocal microscopy. The content of NT, a specific marker of peroxynitrite, was evaluated by Western blotting. The morphological changes of vascular endothelium were observed. Results Compared with control group, the transcription and protein levels of both COX-2 and iNOS increased obviously and their interaction enhanced significantly in model group, in accord with the increased NT content and the pathological alterations of aorta. In ginsenoside group, all these alterations were attenuated significantly ( P < 0.01 or P < 0.05). Conclusions It is proved that there exists interaction between COX-2 and iNOS, aggravating endothelial lesion through peroxynitrite and ginsenoside might antagonize their interaction, playing a protective role.

Autoantibodies directed against phospholipids or human beta 2-glycoprotein I in HIV-seropositive patients: relationship with endothelial activation and antimalonic dialdehyde antibodies.

We investigated the possible role of antiphospholipid (APA) and anti-human 2-glycoprotein I (beta2-GPI) antibodies (Ab) in thrombosis and atherosclerosis in human immunodeficiency (HIV)-positive patients, in whom they seem to be more frequent. We measured APA and anti-beta2-GPI Ab in 58 HIV-positive patients together with markers of disease progression, circulating beta2-GPI, plasma lipids, biological markers of endothelial activation and integrity (plasma thrombomodulin, von Willebrand factor, vascular cell adhesion molecule 1) and with antimalonic dialdehyde antibodies (anti-MDA Ab). We found a 41% frequency of IgG APA in the HIV-positive patients. APA IgMs were rarely positive (7%), and anti-beta2-GPI IgGs were positive in 3-4% patients. There was no correlation between APA or anti-beta2-GPI Ab and the presence of opportunistic infections. Although plasma thrombomodulin, von Willebrand factor and vascular cell adhesion molecule 1 were significantly increased in the HIV-positive patients, APA was correlated only with vascular cell adhesion molecule 1, suggesting that APAs are correlated with endothelial activation but not with vascular endothelial lesions. A correlation between APA and anti-MDA IgG was demonstrated using multivariate analysis (r=0.542, P < 0.0001), suggesting a relationship between the targets of these antibodies. Finally, IgG APAs are frequent in HIV infection but are not correlated with biological markers of endothelial injury. Our results do not support a role for APA or anti-beta2-GPI in HIV-associated silent vascular endothelial damage. However, the role of these autoantibodies in clinically relevant thrombotic events should be investigated in HIV-positive patients.

ELEVATED LEVELS OF TUMOR NECROSIS FACTOR (TNF) IN ADOLESCENTS ON HEMODIALYSIS (HD)

TNF is a cytokine released by activated monocytes exerting a number of reactions that may occur during HD (fever, neutropenia, monocytopenia, vascular endothelial lesions). We measured serum levels of TNF in 6 adolescents aged 14-21 years undergoing regular HD 5 min before and 15 min after start of the procedure with cuprophane membranes. Before HD TNF could not be detected in measurable amounts. In contrast, 5 of 6 sera taken after start of the HD session we found significant amounts of TNF (1045-7000 pg/ml serum). Our results suggest that TNF is released by HD. Therefore the above mentioned side effects of HD may be related to the action of TNF. The high TNF levels may also contribute to the development of autoantibodies or the persistance of cytotoxic antibodies in HD pts.Supported by BMFT, Bonn, FRG, PTB#03 8397