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This study aims to explore the feasibility of a new combined method of constructing an unstable atherosclerotic plaque model in the abdominal aorta of New Zealand rabbits. The experimental New Zealand rabbits were fed adaptively for one week before being punctured through the abdominal aorta with a balloon to injure the intima. Vitamin D (1.5 mL/kg) was administered one week after the surgical model was completed. Following injection, the animals were fed a high-fat diet for 16 weeks. Highresolution magnetic resonance imaging (HR-MRI) and a histology-based method were used to examine the abdominal aortic lesions. The overall mortality rate of the New Zealand rabbit model of unstable atherosclerotic plaque was 17%. The MRIs indicated that the abdominal aorta in the experimental group was eccentrically thickened, while the vascular wall of the abdominal aorta in the control group exhibited no abnormality. The intima of the abdominal aorta in the experimental group showed pathological manifestations of unstable atherosclerotic plaque, while that in the control group exhibited no abnormality. This method of modelling is simple to conduct, has a high success rate and requires a short duration, making it an effective animal model of unstable atherosclerotic plaque. [ABSTRACT FROM AUTHOR]

Background and objectives: Abnormal neointimal hyperplasia (NIH) is known as the predominant mechanism in the pathogenesis of arterial restenosis after balloon angioplasty. Low shear stress (SS) is known to augment balloon injury–induced NIH. The aim of this study is to study the effect and mechanisms of an increase of shear stress caused by arteriovenous fistula could alleviate arterial NIH caused by balloon injury. Methods and results: Eighteen male rabbits were randomly divided into three groups: BI—the rabbits received a balloon injury to right common carotid artery (CCA). BI+AVF—the rabbits received a balloon injury to right CCA and a carotid–jugular AVF. Control—the animals received no surgery. After 21 days, CCA samples were harvested for histological staining, immunohistochemistry, and western blot analysis. The luminal shear stress of the BI+AVF group increased from 13.8 ± 1.0 dyn/cm2 before surgery to 30.9 ± 1.7 dyn/cm2 right after surgery (p < 0.01). This value was higher than that of the BI or Control groups at any timepoint. The neointimal area and neointima/media area ratio in the BI+AVF group were significantly lower than those in the BI group. In the BI group, the cellular proliferation, the protein levels of yes-associated protein (YAP), connective tissue growth factor (CTGF), phospho-c-Jun N-terminal kinase (pJNK), and vascular cell adhesion protein 1 (VCAM1) increased, whereas the protein levels of SMCs specific genes decreased. In the BI+AVF group, the opposite effect was observed as cellular proliferation and the protein levels of YAP, CTGF, pJNK, and VCAM1 decreased, the protein levels of SMCs specific genes increased. Conclusion: The arteriovenous fistula alleviated the balloon injury–induced arterial NIH. It elevated the luminal shear stress and inhibited SMCs phenotypic modulation to the synthetic state, as well as suppressing the over-activation of YAP, JNK, and VCAM1. [ABSTRACT FROM AUTHOR]

Objectives: Impacts of molecular pathways have been discussed recently on restenosis after percutaneous coronary intervention (PCI). Hence, this study aimed to explore the impact of calcineurin‐like phosphoesterase domain containing 1 (CPPED1) and specificity protein 1 (SP1) on restenosis after PCI. Methods: A carotid balloon injury rat model was established, followed by western blot analysis of SP1 and CPPED1 expression in carotid artery (CA) tissues. After SP1 and CPPED1 were overexpressed, the neointimal hyperplasia and luminal stenosis were assessed. In addition, EPC underwent hypoxia/reoxygenation (H/R) treatment to construct an endothelial injury cell model. Then, cell proliferation, apoptosis, intracellular reactive oxygen species (ROS), and Ca2+ concentration were detected with cell counting kit‐8 (CCK‐8), flow cytometry, Chloromethyl‐2'7'‐dichlorofluorescein diacetate (CM‐H2DCFDA) penetrant, and Fluo‐4 AM staining, respectively. The binding relationship between SP1 and CPPED1 was verified by dual‐luciferase reporter and chromatin immunoprecipitation (ChIP) assays. Results: SP1 and CPPED1 were lowly expressed in the model rats with carotid balloon injury. Mechanistically, SP1 bound to the promoter region of CPPED1 to activate CPPED1 expression. Overexpressing SP1 or CPPED1 lowered neointimal formation and restenosis rate, thus promoting the recovery of carotid balloon injury in rats. Meanwhile, SP1 and CPPED1 upregulation reduced ROS levels, Ca2+ concentration, and apoptosis of EPCs, accompanied by accelerated EPC viability. Conclusions: SP1 or CPPED1 overexpression reduced neointimal formation and restenosis rate in carotid balloon injury. [ABSTRACT FROM AUTHOR]

Background: Endovascular treatment of atherosclerotic arterial disease exhibits sex differences in clinical outcomes including restenosis. However, sex-specific differences in arterial identity during arterial remodeling have not been described. We hypothesized that sex differences in expression of the arterial determinant erythropoietin-producing hepatocellular receptor interacting protein (Ephrin)-B2 occur during neointimal proliferation and arterial remodeling. Methods and results: Carotid balloon injury was performed in female and male Sprague–Dawley rats without or 14 days after gonadectomy; the left common carotid artery was injured and the right carotid artery in the same animal was used as an uninjured control. Arterial hemodynamics were evaluated in vivo using ultrasonography pre-procedure and post-procedure at 7 and 14 days and wall composition examined using histology, immunofluorescence and Western blot at 14 days after balloon injury. There were no significant baseline sex differences. 14 days after balloon injury, there was decreased neointimal thickness in female rats with decreased smooth muscle cell proliferation and decreased type I and III collagen deposition, as well as decreased TNFα- or iNOS-positive CD68+ cells and increased CD206− or TGM2-positive CD68+ cells. Female rats also showed less immunoreactivity of VEGF-A, NRP1, phosphorylated EphrinB2, and increased Notch1, as well as decreased phosphorylated Akt1, p38 and ERK1/2. These differences were not present in rats pretreated with gonadectomy. Conclusions: Decreased neointimal thickness in female rats after carotid balloon injury is associated with altered arterial identity that is dependent on intact sex hormones. Alteration of arterial identity may be a mechanism of sex differences in neointimal proliferation after arterial injury. [ABSTRACT FROM AUTHOR]

Intimal hyperplasia (IH) is a common complication of vascular interventional procedures that leads to narrowing of the vessel lumen. 2-Methoxyestradiol (2ME), an estrogen metabolite, has numerous pharmacological actions, including vasoprotective and antiproliferative activities. The present study aimed to evaluate the potential of 2ME, prepared as a self-nanoemulsifying drug delivery system (SNEDDS), to inhibit IH induced by balloon injury (BI) in the rat carotid artery. The prepared 2ME SNEDDS had a particle size of 119 ± 2.3 nm and a zeta potential of −7.1 ± 1.4 mV. Animals were divided into 5 groups, namely control, sham, BI, BI + 2ME (100 μg/kg), and BI + 2ME (250 μg/kg). The obtained data indicated that 2ME significantly inhibited IH as indicated by the histological and morphometric assessment of the intima, media and lumen areas. This was associated with enhanced expression of Bax and inhibited expression of Bcl2 mRNA. Furthermore, 2ME exhibited significant antioxidant properties as evidenced by prevention of malondialdehyde accumulation as well as superoxide dismutase and catalase enzymatic exhaustion. In addition, 2ME showed significant anti-inflammatory actions as it significantly inhibited vascular content of interleukin-6, tumor necrosis factor-alpha, and nuclear factor-κB. The observed vasoprotective activities of 2ME were accompanied by inhibition of Janus kinase/signal transducers and activators of transcription (JAK/STAT) protein expression. In conclusion, this study revealed that 2ME ameliorates balloon injury-induced IH in rats via suppressing JAK/STAT axis. This may help to develop new strategies to combat IH. [ABSTRACT FROM AUTHOR]

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine balloon denudation model, which is comparable with established vessel injury models in large animals. The aorta denudation model with balloon catheters mimics the clinical setting and leads to comparable pathobiological and physiological changes. Briefly, after performing a horizontal incision in the aorta abdominalis, a balloon catheter will be inserted into the vessel, inflated, and introduced retrogradely. Inflation of the balloon will lead to intima injury and overdistension of the vessel. After removing the catheter, the aortic incision will be closed with single stiches. The model shown in this article is reproducible, easy to perform, and can be established quickly and reliably. It is especially suitable for evaluating expensive experimental therapeutic agents, which can be applied in an economical fashion. By using different knockout-mouse strains, the impact of different genes on MH development can be assessed.

The primary challenge in percutaneous coronary interventions for vascular restenosis is the occurrence of restenosis, which is defined by the excessive proliferation of neointimal tissue. Herein, our research team suggests that exosomes obtained from PSC, when paired with quercetin (Q@PSC-E), successfully reduce neointimal hyperplasia in a Sprague-Dawley rat model. Furthermore, the physical properties of the synthesized Q@PSC-E were examined using UV–vis, DLS, and FT-IR characterization techniques. The rats were subjected to balloon injury (BI) utilizing a 2-Fr Fogarty arterial embolectomy balloon catheter. Intimal hyperplasia and the degree of VSMC proliferation were evaluated using histological analysis in the rat groups that received a dosage of Q@PSC-E at 30 mg/kg/d. Significantly, Q@PSC-E inhibited cell proliferation through a pathway that does not include lipoxygenase, as demonstrated by [3H] thymidine incorporation, MTT, and flow cytometry studies. Additionally, the data indicate that Q@PSC-E hinders cell proliferation by targeting particular events that promote cell growth, including the activation of Akt and NF-κB, disruption of cell-cycle progression and also obstructs the ERK signaling pathway. In vascular restenosis, the primary limitation of percutaneous coronary interventions is restenosis, typified by the proliferation of neointimal tissue. Herein, our research team posits that exosomes derived from PSC, combined with quercetin (Q@PSC-E), effectively mitigate neointimal hyperplasia in a Sprague-Dawley rat model. Moreover, the physical characterization of Q@PSC-E was analyzed using UV–vis, DLS, and FT-IR spectroscopy. The balloon injury (BI) in the rats was induced using 2-Fr Fogarty arterial embolectomy balloon catheter. In the rat groups treated with Q@PSC-E (30 mg/kg/d), intimal hyperplasia and the extent of VSMC proliferation were assessed through histopathological analysis. Notably, Q@PSC-E curtailed cell proliferation through a lipoxygenase-independent route, as evidenced by [3H] thymidine incorporation, MTT, and flow cytometry analyses. Additionally, data reveal that Q@PSC-E impedes cell proliferation through specific proliferative events, encompassing Akt and NF-κB activations and the interruption of cell-cycle progression. Following this, it also hinders the ERK signaling pathway. [Display omitted] • Generation of high oxidant species attenuates intimal hyperplasia in balloon injury. • Herein a novel quercetin flavonoid conjugated PSC-exosome was synthesized. • Q@PSC-E successfully hinders cell growth by affecting specific growth-related processes mainly AKT and NF-kB activation. • Histopathological examination confirmed successfully alleviation of restenosis via AKT, ERK and NF-kB signaling pathway. [ABSTRACT FROM AUTHOR]

Xiao Wei,1,* Zhiwei Fang,2,* Jing Sheng,1 Yu Wang,3 Ping Lu1 1Department of Geriatrics, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, People’s Republic of China; 2Engineering Research Center of Cell & Therapeutic Antibody, Ministry of Education, School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, People’s Republic of China; 3Department of Cardiology, Shidong Hospital of Yangpu District, Shanghai 200438, People’s Republic of China*These authors contributed equally to this workCorrespondence: Yu Wang; Ping Lu Email 1300941009@qq.com; pinglushanghai@163.comIntroduction: The main pathological mechanism of restenosis after percutaneous coronary intervention (PCI) is intimal hyperplasia, which is mainly caused by proliferation and migration of vascular smooth muscle cells (VSMCs). Our previous study found that honokiol (HNK), a small-molecule polyphenol, can inhibit neointimal hyperplasia after balloon injury, but its specific mechanism is still unclear. Moreover, poor water solubility as well as low bioavailability of honokiol has limited its practical use.Methods: We used mesoporous silica nanoparticles (MSNPs) as a standard substance to encapsulate HNK and then assemble into honokiol-mesoporous silica nanoparticles, and we investigated the effect of these nanoparticles on the process of restenosis after common carotid artery injury in rats.Results: We report a promising delivery system that loads HNK into MSNPs and finally assembles it into a nanocomposite particle. These HNK-MSNPs not merely inhibited proliferation and migration of VSMCs by reducing phosphorylation of Smad3, but also showed a higher suppression of intimal thickening than the free-honokiol-treated group in a rat model of balloon injury.Conclusion: To sum up, this drug delivery system supplies a potent nano-platform for improving the biological effects of HNK and provides a promising strategy for preventing vascular restenosis.Keywords: mesoporous silica nanoparticles, honokiol, vascular smooth muscle cells, TGF-β pathway, balloon injury, intimal thickening, restenosis

The primary challenge in percutaneous coronary interventions for vascular restenosis is the occurrence of restenosis, which is defined by the excessive proliferation of neointimal tissue. Herein, our research team suggests that exosomes obtained from PSC, when paired with quercetin (Q@PSC-E), successfully reduce neointimal hyperplasia in a Sprague-Dawley rat model. Furthermore, the physical properties of the synthesized Q@PSC-E were examined using UV-vis, DLS, and FT-IR characterization techniques. The rats were subjected to balloon injury (BI) utilizing a 2-Fr Fogarty arterial embolectomy balloon catheter. Intimal hyperplasia and the degree of VSMC proliferation were evaluated using histological analysis in the rat groups that received a dosage of Q@PSC-E at 30 mg/kg/d. Significantly, Q@PSC-E inhibited cell proliferation through a pathway that does not include lipoxygenase, as demonstrated by [ 3 H] thymidine incorporation, MTT, and flow cytometry studies. Additionally, the data indicate that Q@PSC-E hinders cell proliferation by targeting particular events that promote cell growth, including the activation of Akt and NF-κB, disruption of cell-cycle progression and also obstructs the ERK signaling pathway., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: To investigate the effect and potential molecular mechanisms of Dipsacoside B (DB), an herb monomer extracted from Dipsacusasper or Lonicera macranthoides, on the migration and proliferation of vascular smooth muscle cells (VSMCs) and balloon-induced neointimal formation. Methods: In vivo, rat abdominal aorta balloon injury model was utilized to investigate the effect of DB on the neointimal formation. In vitro, cultured VSMCs were used to investigate the effect of DB on Angiotensin-II (Ang-II)-induced migration and proliferation of VSMCs. Western blot and immunofluorescence were used to measure PTEN expression. Results: As compared to vehicle control balloon-injury group, DB treatment significantly inhibited the neointimal formation together up-regulated the expression of phosphatase and tension homolog deleted on chromosome 10 (PTEN). Cell proliferations (MTT and Edu incorporation) assays and wound migration measurement further revealed that treatment with DB significantly blunted Ang-II-induced proliferation and migration potential of VSMCs. Western blot analysis exhibited that DB upregulated the expression of PTEN in vivo and in vitro. Conclusions: DB treatment suppresses the proliferation and migration of VSMCs and reduces neointimal formation by the mechanisms involving regulating the phenotype switch of VSMCs via upregulating PTEN expression.

Valsartan has the potential to attenuate neointimal hyperplasia and to suppress the inflammatory response. This study aimed to evaluate the role of valsartan in neointimal hyperplasia and the toll-like receptor 4 (TLR4)-nitric oxide synthase (NOS) pathway in the balloon-injured rat aorta. Forty-eight Wistar rats were randomly allocated to three groups: sham control (control), balloon-injured group (surgery), and balloon-injured+valsartantreated group (valsartan). Rats were killed at 14 and 28 days after balloon-injury, and then the aortic tissues were collected for morphometric analysis as well as for measurements of the mRNA or protein expression of angiotensin II, angiotensin II type 1 (AT1) receptor, angiotensin II type 2 (AT2) receptor, TLR4, endothelial nitric oxide synthase (eNOS), inducible NOS (iNOS), serine/arginine-rich splicing factor 1(SRSF1) and extracellular signal regulated kinase (ERK). Valsartan at a dose of 20 mg/kg/day markedly decreased neointimal hyperplasia in the aorta of balloon-injured rats, and significantly reduced the mRNA or protein expression of TLR4, AT1 receptor, SRSF1 and phosphorylated-ERK (p-ERK) as well as the aortic levels of iNOS (all p<0.05). Moreover, valsartan increased the eNOS level and AT2 receptor mRNA and protein expression levels (all p<0.05). Valsartan prevented neointimal hyperplasia and inhibited SRSF1 expression and the TLR4-iNOS-ERK-AT1 receptor pathway in the balloon-injured rat aorta. [ABSTRACT FROM AUTHOR]

Vascular remodeling (VR), induced by the massive proliferation and reduced apoptosis of vascular smooth muscle cells (VSMCs), is primarily responsible for many cardiovascular conditions, such as restenosis and pulmonary arterial hypertension. Sodium selenite (SSE) is an inorganic selenium, which can block proliferation and stimulate apoptosis of tumor cells; still, its protective effects on VR remains unknown. In this study, we established rat models with carotid artery balloon injury and monocrotaline induced pulmonary arterial hypertension and administered them SSE (0.25, 0.5, or 1 mg/kg/day) orally by feeding tube for 14 consecutive days. We found that SSE treatment greatly ameliorated the development of VR as evidenced by an improvement of its characteristic features, including elevation of the ratio of carotid artery intimal area to medial area, right ventricular hypertrophy, pulmonary arterial wall hypertrophy and right ventricular systolic pressure. Furthermore, PCNA and TUNEL staining of the arteries showed that SSE suppressed proliferation and enhanced apoptosis of VSMCs in both models. Compared with the untreated VR rats, lower expression of PCNA and CyclinD1, but higher levels of Cleaved Caspase-3 and Bax/Bcl-2 were observed in the SSE-treated rats. Moreover, the increased protein expression of MMP2, MMP9, p-AKT, p-ERK, p-GSK3β and β-catenin that occurred in the VR rats were significantly inhibited by SSE. Collectively, treatment with SSE remarkably attenuates the pathogenesis of VR, and this protection may be associated with the inhibition of AKT and ERK signaling and prevention of VSMC’s dysfunction. Our study suggest that SSE is a potential agent for treatment of VR-related diseases.

Various in vivo laboratory rodent models for the induction of artery stenosis have been established to mimic diseases that include arterial plaque formation and stenosis, as observed for example in ischemic heart disease. Two highly reproducible mouse models - both resulting in artery stenosis but each underlying a different pathway of development - are introduced here. The models represent the two most common causes of artery stenosis; namely one mouse model for each myointimal hyperplasia, and atherosclerosis are shown. To induce myointimal hyperplasia, a balloon catheter injury of the abdominal aorta is performed. For the development of atherosclerotic plaque, the ApoE -/- mouse model in combination with western fatty diet is used. Different model-adapted options for the measurement and evaluation of the results are named and described in this manuscript. The introduction and comparison of these two models provides information for scientists to choose the appropriate artery stenosis model in accordance to the scientific question asked.

The present study was designed to investigate the therapeutic effects of injection of dihydroartemisinin (DHA) into the balloon-injured carotid arteries on balloon injury-induced neointimal formation and to explore whether autophagy is involved in the action of DHA. Percutaneous transluminal balloon angioplasty was performed in Sprague-Dawley rats to induce neointimal formation, immediately after which DHA (100 μmol/l × 1 ml) and/or Rapamycin (1 mg/100 μl), were injected into the balloon-injured carotid arteries. After 14 days, the serum samples and carotid artery tissues were harvested for analysis. Rat aortic vascular smooth muscle cells (VSMCs) were pretreated with DMSO (vehicle), DHA (1, 10, and 100 μmol/l), or 3-methyladenine (3-MA; 10 mM) for 1 h and then stimulated with platelet-derived growth factor-BB (PDGF-BB; 10 ng/ml) for another 24 h. Animal experiments showed that DHA attenuated the balloon injury-induced neointimal formation, inflammation and VSMC phenotypic transition by inhibiting the balloon injury-induced autophagy activation. In vitro results showed that DHA attenuated the PDGF-BB-induced VSMC phenotypic transition, proliferation, and migration by inhibiting the PDGF-BB-induced autophagy activation. Taken together, DHA ameliorates balloon injury-induced neointimal formation through suppressing autophagy. This study provides insights into the development of a drug-eluting stent using DHA. [ABSTRACT FROM AUTHOR]

Objective: To investigate the influence and possible mechanism of 17-Allylamino-17-emethoxy-geldanamycin (17-AAG) on the intimal hyperplasia after carotid artery balloon injury hi rats. Methods: Thirty-six male SD rats were divided into Sham group (n=12). BI group (n=12) and 17-AAG group (n=12) randomly. SD rats were injured using a 2F Fogarty balloon embolectomy catheter. 17-AAG group were disposed by intraperitoneal injecting 17-AAG at a dose of 20 mg/kg2d. After carotid artery balloon injuried 21 days, the damage area of the segment were harvested. HE staining was performed to observe the morphological changes of intima and to evaluate the degree of intimal hyperplasia. The expression of proliferating cell nuclear antigen (PC'NA) hi each group were by imniunohistocheniical staining to evaluate the proliferation of vascular smooth muscle cells. The degree of apoptosis of vascular smooth muscle cells in each group were detected by flow cytometry. Results: The results of HE staining showed that there were different degr ees of intimal hyperplasia following carotid artery injury in BI group and 17-AAG group. I'M ratio of BI group and 17-AAG group significantly increased than that of Sham gr oup (P<0.05). I'M ratio of 17-AAG group distinctly decreased than that of BI group (P<0.05). The results of hmiiunohistochemical staining showed that the expression of PC’NA in carotid artery in BI group and 17-AAG group were significantly higher than that hi Sham group (P<0.05). The expression of PC’NA in 17-AAG group was significantly lower than that in BI group (P<0.05). Compared with Sham gr oup, the apoptosis rate of vascular smooth muscle cells in BI and 17-AAG gr oup significantly increased (P<0.05). Compared with BI gr oup, the apoptosis rate of vascular smooth muscle cells hi 17-AAG group obviously increased (P<0.05). Conclusions: The results of the present study demonstrated that 17-A AG appeared to attenuate intimal hyperplasia after carotid artery balloon injury in rats. The possible mechanism of inhibition of neohithnal formation may be enhance the apoptosis rate of vascular smooth muscle cells. [ABSTRACT FROM AUTHOR]

Vascular restenosis has been proved as the major drawback of percutaneous coronary interventions, which is characterized by neointimal hyperplasia. Naringenin is a kind of natural dihydroflavonoid with a variety of beneficial effects, including anti-oxidative, anti-microbial, anti-cancer and anti-inflammatory properties. However, the effects of naringenin on vascular restenosis remain unclear. This study aimed at investigating the effect and the mechanisms of naringenin on balloon injury (BI)-induced neointimal hyperplasia in the common carotid artery (CCA). BI model of CCA was induced by a 2F Forgarty catheter balloon, and the pathological process of neointimal hyperplasia was noted at 1, 3, 7 and 14 days. Neointimal hyperplasia in CCA increased significantly, especially on day 14 after BI. Subsequently, naringenin (25, 50, 100 mg/kg/d) or volume-matched vehicle were administered to the rats by gavage daily for 14 days. Ultrasound detection and histopathological examination showed that naringenin dose-dependently inhibited BI-induced intimal hyperplasia, as evidenced by reducing imima-media thickness (IMT), neointimal area (NIA), neointimal area/media area (NIA/MA) and neointimal area/internal elastic area (NIA/IELA). Immunohistochemistry revealed that naringenin decreased the expression of proliferating cell nuclear antigen (PCNA) and the cluster of differentiation 163 (CD163). ELISA indicated naringenin significantly reduced the overproduction of IL-1β and TNF-α. By detecting the activity of superoxide dismutase and the level of malondialdehyde and glutathione, we found that naringenin attenuated BI-induced oxidative stress. Additionally, RT-qPCR demonstrated that receptor-interacting protein 1 (RIP1), RIP3 and mixed lineage kinase domain-like (MLKL) mRNA expression were further down-regulated by naringenin treatment. These results suggested that naringenin can suppress BI-induced vascular neointimal hyperplasia through anti-inflammation and anti-oxidative stress, which may be related to the regulation of RIP1-RIP3-MLKL signaling pathway. Image 1 [ABSTRACT FROM AUTHOR]

Introduction: Oral statins reduce intimal hyperplasia (IH) after arterial injury by only ∼25%. Alternative drug delivery systems have gained attention as carriers for hydrophobic drugs. We studied the effects of simvastatin (free vs hyaluronic acid-tagged polysialic acid–polycaprolactone micelles) on vascular smooth muscle cell (VSMC) migration, VSMC proliferation and intimal hyperplasia. We hypothesized both free and micelle containing simvastatin would inhibit VSMC chemotaxis and proliferation, and local statin treatment would be more effective than oral in reducing IH in rats following carotid balloon injury. Methods: VSMCs pretreated with free simvastatin (20 minutes or 20 hours) or simvastatin-loaded micelles underwent chemotaxis and proliferation to platelet-derived growth factor. Next, rats that underwent balloon injury of the common carotid artery received statin therapy—intraluminal simvastatin-loaded micelles prior to injury, periadventitial pluronic gel following injury, or combinations of gel, micelle, and oral simvastatin. After 14 days, morphometric analysis determined the –intimal to medial ratio. Findings were compared to controls receiving oral simvastatin or no statin therapy. Statistical analysis was by analysis of variance for the in vitro experiments and a factorial general linear model for the in vivo experiments. Results: The simvastatin-loaded micelles and free simvastatin inhibited VSMC chemotaxis (54%-60%). IH was induced in all injured vessels. Simvastatin in pluronic gel or micelles reduced IH compared to untreated controls (0.208 ± 0.04 or 0.160 ± 0.03 vs 0.350 ± 0.03, respectively); however, neither gel nor simvastatin-loaded micelles were superior to oral statins (0.261 ± 0.03). Addition of oral statins or combining both local therapies did not provide additional benefit. Micelles were the single greatest contributing factor in IH attenuation. Conclusions: Intraluminally or topically delivered statins reduced IH. The efficacy of single-dose, locally delivered statin alone may lead to novel treatments to prevent IH. The different routes of administration may allow for treatment during endovascular procedures, without the need for systemic therapy. [ABSTRACT FROM AUTHOR]

Objectives: This study aims to evaluate the effects of dihydroartemisinin (DHA) on the balloon injury‐induced neointimal formation in rats and to investigate the underlying mechanism. Methods: The balloon‐induced carotid artery injury model was established in male Sprague–Dawley rats, immediately after which the DHA solution was injected into the tail vein of rats. In in vitro assays, primary rat vascular smooth muscle cells (VSMCs) were pretreated with DHA and then coincubated with LPS. Results: DHA ameliorated the induced neointimal formation and fibrosis but enhanced apoptosis in rat carotid artery after balloon injury. Furthermore, DHA suppressed migration and enhanced apoptosis of the lipopolysaccharide (LPS)‐treated primary VSMCs in vitro. Moreover, in both the balloon injury‐induced rat sera and the LPS‐treated VSMCs, DHA significantly inhibited proinflammatory cytokines, including interleukin‐1β, tumor necrosis factor‐ɑ, and matrix metalloproteinase‐1. Importantly, DHA significantly decreased the balloon injury‐increased expression of nuclear factor kappa B (NF‐κB) subunit NF‐κB p65 expression, and increased the balloon injury‐reduced expression of inhibitor of NF‐κB‐alpha, indicating the inhibition of the IκB/NF‐κB pathway. Conclusion: DHA significantly inhibited neointimal formation in balloon‐induced rat carotid artery injury and the mechanism may be related to the inhibition of IκB/NF‐κB signaling, which alleviates the inflammatory response. Dihydroartemisinin (DHA) significantly inhibited neointimal formation in balloon‐induced rat carotid artery injury and the mechanism may be related to the inhibition of inhibitor of NF‐κB (IκB)/nuclear factor kappa B (NF‐κB) signaling, which alleviates the inflammatory response. [ABSTRACT FROM AUTHOR]

Background/Aims: Soluble guanylate cyclase (sGC) exists as reduced, oxidized, and heme-free forms. Currently, it is unclear whether endovascular mechanical stenosis has an impact on vascular tone control by drugs targeting sGC, namely cGMP generators. Methods: Pharmacological responses to acidified sodium nitrite (reduced sGC stimulant) and BAY 60-2770 (oxidized/heme-free sGC stimulant) were studied in balloon-injured rat carotid arteries at several time points. In addition, sGC expression was detected by immunohistochemistry. Results: At 1 day after injury, acidified sodium nitrite-induced relaxation was attenuated in the injured artery, whereas BAY 60-2770-induced relaxation was augmented. Similar attenuation of response to acidified sodium nitrite was seen at 7 and 14 days after injury. On the other hand, the augmentation of response to BAY 60-2770 disappeared at 7 and 14 days after injury. At 1 day after injury, the immunohistochemical expression pattern of sGC in the smooth muscle layer of the injured artery was not different from that of the uninjured artery. However, in the injured artery, the intensity of sGC staining was weak at 7 and 14 days after injury. Conclusion: Balloon injury alters vascular responsiveness to cGMP generators, which seems to be associated with the form and/or expression of sGC. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • TSG inhibited intimal hyperplasia in rat common carotid artery balloon injury. • TSG reduced the expression of SCF/c-kit, SDF-1α/CXCR4 and FKN/CX3CR1 axes. • TSG enhanced re-endothelialization through increasing VEGF levels and CD34 cells. Abstract Background Intimal hyperplasia is the major therapeutic concern after percutaneous coronary intervention. The aim of this study is to investigate effects of 2,3,4′,5-tetrahydroxystilbene-2-O-β-D glucoside (TSG) on intimal hyperplasia and the underling mechanisms through attenuating the expressions of stromal cell-derived factor-1α (SDF-1α)/CXCR4, stem cell factor (SCF)/c-kit and fractalkine (FKN)/CX3CR1, and through promoting re-endothelialization with vascular endothelial growth factor (VEGF). Method Rats were operated with carotid artery balloon injury. The treatment groups were gavaged with 50 and 100 mg/kg/d of TSG. After 10 days of treatment, carotid artery pathological changes were evaluated by histology. Serum levels of SDF-1α, SCF, FKN and VEGF were detected by enzyme linked immunosorbent assay. The protein expressions of the receptors c-kit, CXCR4, CX3CR1, as well as CD34 and proliferating cell nuclear antigen (PCNA) were detected by immunochemistry. Results TSG dose-dependently inhibited balloon injury-induced intimal hyperplasia, as evidenced by reducing neointima area (NIA), neointima area/media area (NIA/MA), neointima area/internal elastic area (NIA/IELA), and by decreasing the protein expression of PCNA. TSG reduced serum levels of SDF-1α, SCF and FKN, and it also decreased the expressions of the corresponding receptors c-kit, CXCR4, CX3CR1 in neointima. Importantly, the level of VEGF in peripheral blood and the expression of CD34 in vascular walls were increased to promote re-endothelialization. Conclusions This study clearly demonstrated that TSG was effective in inhibiting intimal hyperplasia, and this effect was mediated, at least in part, through the SCF/c-kit, SDF-1α/CXCR4 and FKN/CX3CR1 axes. Importantly, TSG could increase VEGF and CD34 to promote endothelial repair. [ABSTRACT FROM AUTHOR]

Restenosis after angioplasty is caused usually by neointima formation characterized by aberrant vascular smooth muscle cell (VSMC) dedifferentiation. Myeloid-derived growth factor (MYDGF), secreted from bone marrow-derived monocytes and macrophages, has been found to have cardioprotective effects. In this study we investigated the effect of MYDGF to postinjury neointimal formation and the underlying mechanisms. Rat carotid arteries balloon-injured model was established. We found that plasma MYDGF content and the level of MYDGF in injured arteries were significantly decreased after balloon injury. Local application of exogenous MYDGF (50 μg/mL) around the injured vessel during balloon injury markedly ameliorated the development of neointimal formation evidenced by relieving the narrow endovascular diameter, improving hemodynamics, and reducing collagen deposition. In addition, local application of MYDGF inhibited VSMC dedifferentiation, which was proved by reversing the elevated levels of osteopontin (OPN) protein and decreased levels of α-smooth muscle actin (α-SMA) in the left carotid arteries. We showed that PDGF-BB (30 ng/mL) stimulated VSMC proliferation, migration and dedifferentiation in vitro; pretreatment with MYDGF (50-200 ng/mL) concentration-dependently eliminated PDGF-BB-induced cell proliferation, migration and dedifferentiation. Molecular docking revealed that MYDGF had the potential to bind with sphingosine-1-phosphate receptor 2 (S1PR2), which was confirmed by SPR assay and Co-IP analysis. Pretreatment with CCG-1423 (Rho signaling inhibitor), JTE-013 (S1PR2 antagonist) or Ripasudil (ROCK inhibitor) circumvented the inhibitory effects of MYDGF on VSMC phenotypic switching through inhibiting S1PR2 or its downstream RhoA-actin monomers (G-actin) /actin filaments (F-actin)-MRTF-A signaling. In summary, this study proves that MYDGF relieves neointimal formation of carotid arteries in response to balloon injury in rats, and suppresses VSMC dedifferentiation induced by PDGF-BB via S1PR2-RhoA-G/F-actin-MRTF-A signaling pathway. In addition, our results provide evidence for cross talk between bone marrow and vasculature., (© 2023. The Author(s), under exclusive licence to Shanghai Institute of Materia Medica, Chinese Academy of Sciences and Chinese Pharmacological Society.)

Ginsenoside Re (GS-Re) is one of the main ingredients of ginseng, a widely known Chinese traditional medicine, and has a variety of beneficial effects, including vasorelaxation, antioxidative, anti-inflammatory, and anticancer properties. The aims of the present study were to observe the effect of GS-Re on balloon injury-induced neointimal hyperplasia in the arteries and to investigate the mechanisms underlying this effect. A rat vascular neointimal hyperplasia model was generated by rubbing the endothelium of the common carotid artery (CCA) with a balloon, and GS-Re (12.5, 25 or 50 mg/kg/d) were subsequently continuously administered to the rats by gavage for 14 days. After GS-Re treatment, the vessel lumen of injured vessels showed significant increases in the GS-Re 25.0 and 50.0 mg/kg/d (intermediate- and high-dose) groups according to H.E. staining. Additionally, a reduced percentage of proliferating cell nuclear antigen (PCNA)-positive cells and an increased number of SM α-actin-positive cells were detected, and the levels of NO, cyclic guanosine monophosphate (cGMP), and eNOS mRNA as well as the phos-eNOS ser1177 /eNOS protein ratio were obviously upregulated in the intermediate- and high-dose groups. Moreover, the promotive effects of GS-Re on NO and eNOS expression were blocked by L-NAME treatment to different degrees. These results suggested that GS-Re can suppress balloon injury-induced vascular neointimal hyperplasia by inhibiting VSMC proliferation, which is closely related to the activation of the eNOS/NO/cGMP pathway. [ABSTRACT FROM AUTHOR]

Hepatic growth factor (HGF) has been widely used in studies on arterial remodeling after injury, and results turn out to be inconsistent. The changes of endogenous HGF expression after injury also remain controversial. This study clarified the role of exogenous human HGF (hHGF) gene transfer in neointimal hyperplasia and investigated the associated alterations of endogenous HGF and c-Met expressions under endothelial denudation with or without hHGF gene transfer using a balloon-injured rabbit aorta model. Sixty-one rabbits were randomly divided into normal controls, endothelial injury, endothelial injury with hHGF, or the control vector gene transfer groups. On weeks 1, 2, 4, and 8 after injury, neointimal hyperplasia and endothelialization were evaluated by the ratio of neointimal area to medial area (N/M ratio), CD31-positive staining, α-smooth muscle actin, and endothelial nitric oxide synthase expressions using histological analysis, immunohistochemistry staining, or real-time quantitative reverse transcriptase polymerase chain reaction. Endogenous rabbit HGF (rHGF) and c-Met expressions were detected with immunohistochemistry staining and quantitative reverse transcriptase polymerase chain reaction. It was found that expressions of endogeneous rHGF and c-Met in endothelial injury upregulated with peak levels on week 2 or week 4 after injury ( p  < 0.01). On week 1 after hHGF transfer, neointimal hyperplasia was significantly inhibited ( p  < 0.001), with decreased α-smooth muscle actin expression ( p  < 0.05) and improved endothelial cells regeneration and function ( p  < 0.01). More remarkable overexpression of endogenous rHGF and c-Met mRNAs were detected, and lowered positive staining of rHGF and c-Met was shown in the neointima ( p  < 0.05). These results demonstrated hHGF gene transfer induced further overexpression of endogenous rHGF and c-Met mRNAs but lowered immunoreactivities of rHGF and c-Met in the neointima, thus leading to significant attenuation of neointimal hyperplasia. [ABSTRACT FROM AUTHOR]

Intimal hyperplasia is the main cause of restenosis after carotid artery injury, and the underlying mechanism involves the proliferation and migration of vascular smooth muscle cells (VSMCs). Angiotensin II Type 1 Receptor-Associated Protein (ATRAP) has been reported to withstand intimal hyperplasia by inhibiting VSMCs proliferation and migration; however, whether the beneficial effect of ATRAP associates with VSMCs apoptosis remains unclarified. We demonstrated that the adenoviral-mediated overexpression of ATRAP induced VSMC apoptosis, alleviating the balloon injury-induced neointima formation in rats. Under the condition of Angiotensin-II stimulation, ATRAP overexpression induced the apoptosis of rat VSMCs by depressing the PI3K-Akt signaling; whereas up-regulation of Akt by PTEN inhibitor abolished the apoptotic death. Thus, ATRAP regulates carotid intimal hyperplasia through controlling the PI3K-Akt signal-mediated VSMCs apoptosis. [ABSTRACT FROM AUTHOR]

The aims of the present study were to determine the effects of heparin-derived oligosaccharides (HDOs) on vascular intimal hyperplasia (IH) in balloon-injured carotid artery and to elucidate the underlying mechanisms of action. An animal model was established by rubbing the endothelia within the common carotid artery (CCA) in male rabbits. The rabbits were fed a high-cholesterol diet. Arterial IH was determined by histopathological changes to the CCA. Serum lipids were detected using an automated biochemical analysis. Expressions of mRNAs for vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF), vascular cell adhesion molecule-1 (VCAM-1), monocyte chemoattractant protein-1 (MCP-1), scavenger receptor class B type I (SR-BI), and ATP-binding cassette transporter A1 (ABCA-1) were analyzed using reverse transcription polymerase chain reaction assays. Expressions of VEGF, VCAM-1, MCP-1, SR-BI and ABCA-1 proteins were analyzed by Western blotting. Enzyme-linked immunosorbent assays were used to quantify expression levels of VEGF and bFGF. Our results showed that administration of HDO significantly inhibited CCA histopathology and restenosis induced by balloon injury. The treatment with HDOs significantly decreased the mRNA and protein expression levels of VEGF, bFGF, VCAM-1, MCP-1, and SR-BI in the arterial wall; however, ABCA-1 expression level was elevated. HDO treatment led to a reduction in serum lipids (total cholesterol, triglycerides, high-density and low-density lipoproteins). Our results from the rabbit model indicated that HDOs could ameliorate IH and underlying mechanism might involve VEGF, bFGF, VCAM-1, MCP-1, SR-BI, and ABCA-1. [ABSTRACT FROM AUTHOR]

Percutaneous transluminal angioplasty is frequently used in patients with severe arterial narrowing due to atherosclerosis. However, it induces severe arterial injury and an inflammatory response leading to restenosis. Here, we studied a potential activation of the endocannabinoid system and the effect of FA amide hydrolase (FAAH) deficiency, the major enzyme responsible for endocannabinoid anandamide degradation, in arterial injury. We performed carotid balloon injury in atherosclerosis-prone apoE knockout (apoE−/−) and apoE−/−FAAH−/− mice. Anandamide levels were systemically elevated in apoE−/− mice after balloon injury. ApoE−/−FAAH−/− mice had significantly higher baseline anandamide levels and enhanced neointima formation compared with apoE−/− controls. The latter effect was inhibited by treatment with CB1 antagonist AM281. Similarly, apoE−/− mice treated with AM281 had reduced neointimal areas, reduced lesional vascular smooth-muscle cell (SMC) content, and proliferating cell counts. The lesional macrophage content was unchanged. In vitro proliferation rates were significantly reduced in CB1−/− SMCs or when treating apoE−/− or apoE−/−FAAH−/− SMCs with AM281. Macrophage in vitro adhesion and migration were marginally affected by CB1 deficiency. Reendothelialization was not inhibited by treatment with AM281. In conclusion, endogenous CB1 activation contributes to vascular SMC proliferation and neointima formation in response to arterial injury.