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Objective:To detect the characteristics of vascular remodeling after carotid balloon injury model in rats using ultrasound biomicroscopy(UBM), and to discuss the application value of UBM technique by comparing ultrasonic characteristics with histopathological results.Methods:Carotid balloon injury was performed in 10-week-old SD rats(11 female and 11 male) by 2F Fogarty balloon catheter. The left common carotid artery(CCA) was injured and the right side in the same animal was used as an uninjured control. Arterial structures and hemodynamics were evaluated pre-procedure and post-procedure at 7, 14 days.The intima-media thickness(IMT) inner diameter, outer diameter, lumen area, vessel area, peak systolic velocity, end diastolic velocity of CCA were measured by UBM, and the vascular resistance index, shear stress and blood flow were calculated to evaluate the vascular hemodynamics. The histological data were obtained by H&E staining in cross-sections at 14 days after balloon injury. The characteristics of arterial structure and hemodynamic changes at various time points were compared, the structural changes of CCA between injured and control side after injury were compared. The Spearman correlation and linear regression were used to test the correlation between ultrasonic and histological measurements 14 days after balloon injury.Results:①Compared with pre-procedure, the IMT at 14 days after balloon injury was increased, the inner diameter was decreased, the shear stress in ultrasound was increased(all P<0.05). H&E staining histological test showed that IMT and neointima area in male rats were larger than those of female rats (all P<0.001). ②After carotid balloon injury, the lumen area decreased, but the CCA underwent compensatory positive remodeling and the vessel area increased. ③Significant correlations were demonstrated between UBM and histology in IMT, inner diameter, outer diameter and vessel area of CCA( rs=0.819, 0.965, 0.896, 0.955; all P<0.001). The vessel area value measured by UBM was larger than that of histology( P=0.006). Conclusions:The CCA of rats can be showed clearly by UBM in males and females. The arterial structure cab be measured by UBM accurately with good correlation with histology, as did arterial hemodynamic parameters, which may be benefit for the study in carotid balloon injury model of rats.
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<p><b>OBJECTIVE</b>To investigate the effect and potential mechanisms of rutaecarpine (Rut) in a rat artery balloon-injury model.</p><p><b>METHODS</b>The intimal hyperplasia model was established by rubbing the endothelia with a balloon catheter in the common carotid artery (CCA) of rats. Fifty rats were randomly divided into five groups, ie. sham, model, Rut (25, 50 and 75 mg/kg) with 10 rats of each group. The rats were treated with or without Rut (25, 50, 75 mg/kg) by intragastric administration for 14 consecutive days following injury. The morphological changes of the intima were evaluated by hematoxylin-eosin staining. The expressions of proliferating cell nuclear antigen (PCNA) and smooth muscle (SM) α-actin in the ateries were assayed by immunohistochemical staining. The mRNA expressions of c-myc, extracellular signal-regulated kinase 2 (ERK2), MAPK phosphatase-1 (MKP-1) and endothelial nitric oxide synthase (eNOS) were determined by real-time reverse transcription-polymerase chain reaction. The protein expressions of MKP-1 and phosphorylated ERK2 (p-ERK2) were examined by Western blotting. The plasma contents of nitric oxide (NO) and cyclic guanosine 3',5'-monophosphate (cGMP) were also determined.</p><p><b>RESULTS</b>Compared with the model group, Rut treatment significantly decreased intimal thickening and ameliorated endothelial injury (P<0.05 or P<0.01). The positive expression rate of PCNA was decreased, while the expression rate of SM α-actin obviously increased in the vascular wall after Rut (50 and 75 mg/kg) administration (P<0.05 or P<0.01). Furthermore, the mRNA expressions of c-myc, ERK2 and PCNA were downregulated while the expressions of eNOS and MKP-1 were upregulated (P<0.05 or P<0.01). The protein expressions of MKP-1 and the phosphorylation of ERK2 were upregulated and downregulated after Rut (50 and 75 mg/kg) administration (P<0.05 or P<0.01), respectively. In addition, Rut dramatically reversed balloon injury-induced decrease of NO and cGMP in the plasma (P<0.05 or P<0.01).</p><p><b>CONCLUSION</b>Rut could inhibit the balloon injury-induced carotid intimal hyperplasia in rats, possibly mediated by promotion of NO production and inhibiting ERK2 signal transduction pathways.</p>
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Animals , Male , Actins , Metabolism , Carotid Arteries , Metabolism , Pathology , Carotid Artery Injuries , Drug Therapy , Genetics , Pathology , Cyclic GMP , Blood , Disease Models, Animal , Gene Expression Regulation , Hyperplasia , Indole Alkaloids , Pharmacology , Therapeutic Uses , Nitric Oxide , Blood , Phosphorylation , Proliferating Cell Nuclear Antigen , Metabolism , Quinazolines , Pharmacology , Therapeutic Uses , RNA, Messenger , Genetics , Metabolism , Rats, Sprague-Dawley , Tunica Intima , PathologyABSTRACT
OBJECTIVE: To study the effects of ginsenoside Rg3 on NF-κBp65 and its related inflammatory factors after carotid balloon injury in rats. METHODS: Sprague Dawley(SD) rats(n=40) were divided into 4 groups randomlysham operationgroup, model group, ginsenoside Rg3 5 mg·kg-1 and ginsenoside Rg3 10 mg·kg-1. 2.0 mm×12 mm Ballon catheters were used to establish the carotid artery intima injury model. On next day after modeling, all animals in model group were administered intragastrically with relative saline and different concentration of ginsenoside Rg3 for 14 d. After 14 d, the morphological changes of the injured arteries were observed by HE staining. The expression of NF-κBp65 in vascular tissue was detected by Western blot; Tumor necrosis factor-α(TNF-α), interleukin-1β(IL-1β) and IL-6 were detected by RT-PCR and Elisa. RESULTS: Compared with the sham-operation group, the neointimal in model group was significantly thicker(P<0.01). while the expression levels of IL-1β, IL-6, TNF-α and NF-κBp65 were increased in model group(P<0.01). The vascular intimal hyperplasia was alleviated distinctly(P<0.01) and the protein expression of NF-κBp65 in vascular tissue was significantly decreased(P<0.01) compared with model group. the expression level of IL-1β, IL-6 and TNF-α was significantly lower in intervention group(P<0.01). CONCLUSION: Ginsenoside Rg3 could reduce the vascular neointimal hyperplasia and inflammation induced by balloon injury, which may be related to its inhibitory role of the transcription factor NF-κB signaling pathway.
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Objective To clarify the role of transmembrane protein 66(TMEM66) in intima proliferation of carotid artery induced by balloon injury in rats.Methods SD rats were randomly divided into three groups (n=10), including control group, the left carotid artery balloon injury group and injury group added with lentiviruses, respectively.Accordingly, the intima proliferation of carotid artery were detected by H/E staining;the expressions of TMEM66 in carotid artery injuried by balloon were determined by Western blot,q-PCR and immunohistochemistry, and the migration and proliferation of VSMCs were measured by scratch test and CCK8 respectively.Results Compared with control group, the expression of TMEM66 in carotid artery was obviously decreased after balloon injury (P<0.05).Most importantly, the intima proliferation of carotid artery was significantly reversed by overexpression of TMEM66 via specific lentiviruses (P<0.05).Conclusions TMEM66 is able to attenuate the intima proliferation of carotid artery after balloon injury.It could be that upregulation of TMEM66 can alleviate the migration and proliferation of VSMCs by PDGF.
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[Objective]To study the effects of Caspase-1 specific inhibitor AC-YVAD-CMK on intimal hyperplasia after carotid artery balloon injury in rats and its possible mechanism.[Methods]A total of 33 male adult SD rats were randomly divided into sham group,balloon injury group and balloon injury+AC-YVAD-CMK group. Using the method of balloon injury to establish rat carotid ar?tery intimal hyperplasia animal model,rats were sacrificed and blood vessels were harvested 14 days after operation. Fifteen vascular segments embedded in OCT and the intima to media(I/M)area ratio of neointima was measured by hematoxylin-eosin(HE)staining;18 vascular segments were harvested and the expression of NLRP3 inflammasome,cleaved-Caspase-1,interleukin(IL)-1βand IL-18 were measured by Western blot.[Results]HE staining showed that AC-YVAD-CMK significantly inhibited the degree of intimal hyperplasia compared with the balloon injury group[(0.78 ± 0.13)vs(1.52 ± 0.14);P=0.000]. The expression of NLRP3 inflamma?some was increased in balloon injury group while the AC-YVAD-CMK attenuates the expression of NLRP3(P=0.009);The expres?sion of cleaved-Caspase-1 was in line with the expression of NLRP3(P=0.000). The levels of pro-inflammatory cytokines IL-1βand IL-18 in balloon injury+AC-YVAD-CMK group were significantly lower than those in the balloon injury group(P=0.000).[Conclusion]AC-YVAD-CMK can attenuate intimal hyperplasia after balloon injury of carotid artery in rats,which might be related to its effect on inhibiting the activation of Caspase-1,which could affect the release of pro-inflammatory cytokine of IL-1βand IL-18.
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Objective To investigate the inhibitory effect of a new antihypertensive drug on carotid artery intimal hyperplasia in balloon-injured rats and its possible mechanism.Methods A total of 48 SD rats were randomly divided as follows:sham operation group,model group,low-dose Azilsartan medoxomil group and high-dose Azilsartan medoxomil group(n =12).The rats in sham operateion group and model group were treated by by gavage with 3ml normal saline every day,and rats in low-dose Azilsartan medoxomil group and high-dose Azilsartan medoxomil group were treated by by gavage at adose of 2mg/kg and 4mg/kg Azilsartan medoxomil(3ml) every day.14 and 28 days after operation,we can observe the morphological changes of the injured arteries by HE staining and measure intimal area (IA),medial area(MA),the intimal/medial area ratio (IA/MA).The expression of PCNA,TLR4,NF-κB p65 in each group were detected by immunohistochemistry and the plasma level of TNF-α,IL-6 were detected by enzyme-linked immunosorbent assay (ELISA).Results Compared with the model group,the intimal area and the intimal / medial area ratio were significantly reduced in the low-dose group and hige-dose group (P ＜ 0.05).The expression of PCNA,TLR4 and NF-κB p65 in carotid artery were significantly lower (P ＜ 0.05).The plasma level of TNF-α、IL-6 were significantly lower (P ＜ 0.05).Compared with the sham operation group,the intimal area and the intimal/medial area ratio were significantly increased in the operation group (P ＜ 0.05) and the expression of PCNA,TLR4 and NF-κB p65 in carotid artery were significantly increased (P ＜0.05).The plasma level of TNF-α、IL-6 were significantly increased (P ＜ 0.05).Conclusion Azilsartan medoxomil can alleviate the incidence of vascular restenosis.The mechanism of action possibly was related to reducing the expression of inflammatory factors such as TNF-α,IL-6 by TLR4/NF-κB pathway and inhibiting the carotid artery intimal hyperplasia.
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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.
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Animals , Male , Rabbits , ATP Binding Cassette Transporter 1 , Carotid Artery Injuries , Drug Therapy , Pathology , Chemokine CCL2 , Heparin , Therapeutic Uses , Hyperplasia , Oligosaccharides , Therapeutic Uses , Tunica Intima , Pathology , Vascular Cell Adhesion Molecule-1 , Vascular Endothelial Growth Factor AABSTRACT
Objective: To investigate the effects of asragaloside IV (AS-IV) on cultured vascular smooth muscle cells (VSMCs) and neointima hyperplasia in the carotid artery of rats after ballon injury and explore the inhibitory mechanisms. Methods: For next related researches, primary cultures of VSMCs were prepared from the thoracoabdominal aorta of rats using explant method. Taking the recombinant rat tumor necrosis factor (TNF-α, 100 ng/mL ) as the stimulating factor, the model of VSMCs proliferation was established by TNF-α inducer in vitro and the effects of AS-IV (0, 0.5, 5, 25, and 50 μg/mL) on the VSMCs proliferation induced by TNF-α were determined by CCK-8 method. Rat carotid artery balloon injury model was prepared by Fogarty (2F) balloon catheter. Fifty healthy male Sprague-Dawley rats were randomly divided into five groups: a Sham-operation group (Sham), a model group (model), and three AS-IV-treated (20, 40, and 60 mg/kg) groups. Three days before the surgery, 1% CMC, AS-IV 20, 40, and 60 mg/kg were ig administered to each group once daily for continuous 17 d. Fifteen days after the surgery, rats were killed, and the carotid arterys were harvested. Hematoxylin-elsin staining was carried out to observe the pathomorphological change in vascular intima. The measurement of lumen area, intimal area, intimal area/medium film area were measured by computer image analysis system; Immunohistochemistry staining was performed to measure the expression of proliferating cell nuclear antigen (PCNA) and platelet-derived growth factor-BB (PDGF-BB). Results: The proliferative activity of VSMCs was obviously increased in the TNF-α group under the stimulation of TNF-α. There was a significant difference compared with the control group (P < 0.01). However, when pretreated with AS-IV ahead of time, we found that AS-IV significantly inhibited TNF-α-induced VSMCs proliferation in a dose- and time-dependent manner compared to the TNF-α group. Compared with the model group, the area of intima, the ratio of intima to media (I/M), and the expression of PCNA and PDGF-BB decreased significantly (P < 0.05) in AS-IV groups. Conclusion: AS-IV exerts the inhibitory effects on VSMCs proliferation in a dose- and time-dependent manner. AS-IV significantly inhibits the neointimal hyperplasia in rat carotid artery. It might be partially attributed to the inhibition of proliferation. It may be one of the mechanisms of inhibiting the proliferation of carotid intima in rats induced by balloon injury that AS-IV inhibits the expression of growth factor PDGF-BB.
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Objective To quickly establish the To quickly establish the atherosclerosis (AS) model in apoprotein E gene knockout (ApoE-/-) rats by short-term high-fat diet combined with balloon injury. Methods Eight-week-old male SD and ApoE-/- rats (10 each) were used in the experiments. After fed with high-fat diet for 2 weeks, the blood routine, hepatorenal function, blood lipid profile and C-reactive protein (CRP) were measured, and then all the left common carotid arteries were processed by balloon injury. After 2 weeks, rats euthanasia were carried out by excessive chloral hydrate, and then the operation side common carotid arteries were stained with HE, Masson trichrome and oil red O. While the common carotid arteries accepted CD68, α-SMA immunofluorescent staining. Results The blood lipid level was significantly higher in ApoE-/- rats than in SD rats [total cholestrol (TC): 18.56±2.82mmol/L vs 5.69±1.98mmol/L, P<0.01; low density lipoprotein (LDL): 6.86±1.47mmol/L vs 1.92±0.76mmol/L, P<0.01]. In the condition of serious blood lipid disorders, the ApoE-/- rats had been in a state of inflammation [CRP: 4.66±0.57mg/L vs 0.39±0.21mg/L, P<0.05; white blood cell: (21.79±5.10)×109/L vs (14.82±2.41)×109/L, P<0.01; neutrophil: (9.28±3.35)×109/L vs (2.10±0.96)×109/L, P<0.01]. HE and Masson staining showed that obvious hyperplasia formed and collagen fibers deposited slightly in the two groups. Oil red O staining revealed the obvious hyperplastic plaques in ApoE-/- rats, but a negative result in SD rats. Immunofluorescence staining showed the significant positive CD68 and weak positive α-SMA in the plaque of ApoE-/- rats, while in SD rats the positive -SMA was pointed out with no CD68 coloration. Conclusion The atherosclerosis model of ApoE-/- rat may be quickly established by short-term high-fat diet combined with balloon injury.
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PURPOSE: Appropriate animal models of atherosclerotic plaque are crucial to investigating the pathophysiology of atherosclerosis, as well as for the evaluation of the efficacy and safety of vascular devices. We aimed to develop a novel animal model that would be suitable for the study of advanced atherosclerotic lesions in vivo. MATERIALS AND METHODS: Atherosclerotic plaque was induced in 24 iliac arteries from 12 rabbits by combining a high cholesterol diet, endothelial denudation, and injection into the vessel wall with either saline (n=5), olive oil (n=6), or inflammatory proteins [n=13, high-mobility group protein B1 (HMGB1) n=8 and tumor necrosis factor (TNF)-α n=5] using a Cricket™ Micro-infusion catheter. Optical coherence tomography (OCT) was performed to detect plaque characteristics after 4 weeks, and all tissues were harvested for histological evaluation. RESULTS: Advanced plaque was more frequently observed in the group injected with inflammatory proteins. Macrophage infiltration was present to a higher degree in the HMGB1 and TNF-α groups, compared to the oil or saline group (82.1±5.1% and 94.6±2.2% compared to 49.6±14.0% and 46.5±9.6%, p-value<0.001), using RAM11 antibody staining. On OCT, lipid rich plaques were more frequently detected in the inflammatory protein group [saline group: 2/5 (40%), oil group: 3/5 (50%), HMGB1 group: 6/8 (75%), and TNF-α group: 5/5 (100%)]. CONCLUSION: These data indicate that this rabbit model of atherosclerotic lesion formation via direct injection of pro-inflammatory proteins into the vessel wall is useful for in vivo studies investigating atherosclerosis.
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Animals , Male , Rabbits , Cholesterol, Dietary/administration & dosage , Disease Models, Animal , Endothelium/surgery , HMGB1 Protein/adverse effects , Iliac Artery/diagnostic imaging , Injections, Intra-Arterial , Macrophages , Olive Oil/adverse effects , Plaque, Atherosclerotic/chemically induced , Sodium Chloride/adverse effects , Tomography, Optical Coherence , Tumor Necrosis Factor-alpha/adverse effectsABSTRACT
Objective To compare the effects of forming atherosclerosis by conducting ballon injury operation after 1th, 2th and 3th week of Vitamin D3(VD3) i.p., exploring the best method for atherosclerosis modeling .Methods 36 male rats were selected for balloon-injured carotid artery .SD rats were divided into 4 groups randomly:control group ( n=6), Model group1 (n=10), Model group2(n=10), Model group3 (n=10).Control group were fed up with common diet.Model groups were fed up with high-fat diet and injected 4.0 ×105 IU/kg VD3 through enterocoelia in the beginning , followed by the balloon-injured left carotid artery operation after 1th, 2th and 3th week respectively and 1.0 ×105 IU/kg VD3injection at 0th, 2th week after operation.The rats were killed at 4th week after operation.The serum levels of TG, TC, HDL-C and LDL-C were checked .ELISA was used to detect the content of hsCRP , IL-6 and TNFα.HE staining was used to observe the pathological changes in the thoracic aorta , and the thoracic aorta thickness , plaque area ( PA) , cross-sectional area of vessel ( CVA) and the ratio of PA to CVA ( PA/CVA) were analyzed .Results After 4 weeks of operation , levels of TC and LDL-C were significantly increased in Model group 2 and 3 compared with that of the control group ( P<0.05).Furthermore, contents of hsCRP, IL-6 and TNFαof model groups were also seriously higher than that of the control group (P<0.05), and that of Model group 3 were the highest.Typical AS plagues were observed in different degrees in model groups, and thoracic aorta thickness and PA/CVA were obviously increased than that of control group (P<0.05). Model group 3 turned out masses of lipid foam cells accumulated , and PA, CVA and PA/CVA were significantly increased than that of Model group2 or 3.Conclusion The AS model can be established successfully in rats with ballon injury after 3 weeks of high-fat diet plus VD3 i.p., which is the ideal method to induced atherosclerosis model .
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OBJECTIVE: To investigate the inhibitory effect of ginsenoside Re on vascular neointimal hyperplasia induced by balloon-injury and probe its molecular mechanism in rats. METHODS: The rat carotid artery neointimal hyperplasia model was established by rubbing the endothelia with a balloon in male Sprague-Dawley rats, then animals were intrapritoneally injected with distilled water in model group and sham operation group or with ginsenoside Re 6, 12 and 24 mg·kg-1·d-1 in other endothelia rubbed groups. After 14 consecutive days, the injuried artery was taken for H&E staining, the histopathological observation and detecting the neointimal area as well as the ratio of neointimal area/media area were taken to evaluate the vescular intimal hyperplasia level. For probing the molecular mechanism, the expression of mitogen-activated protein kinase phosphatase-1 (MKP-1) was detected at the transcript levels by real time RT-PCR, and the protein expressions of MKP-1 and phosphorylation extracellular signal-regulated kinase 1/2 (pERK12) were examined by immunohistochemistry and analyzed with Image-Pro Plus. RESULTS: Compared with the endothelia rubbing model group, ginsenoside Re 6, 12, 24 mg·kg-1·d-1 medications significantly improved the histopathological changes induced by baloon-injury, decreased the elevated neointimal area and the ratio of neointimal area/media area induced by baloon-injury; ginsenoside Re administration could also down-regulate the elevated pERK1/2 protein expression, and significantly up-regulated the decreased MPK-1 mRNA and protein expressions induced by baloon-injury. CONCLUSION: Ginsenoside Re inhibits the vascular neointimal hyperplasia induced by baloon-injury in rats, the molecular mechanism is related to its inhibition effect on ERK signaling.
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OBJECTIVE: To examine the inhibitory effects of evodiamine on vascular neointimal hyperplasia in balloon-injured carotid artery of rats and explore the possible mechanisms. METHODS: Healthy male Sprague Dawley rat vascular neointimal hyperplasia model was made by rubbing the endothelia of common carotid artery with a balloon, and then the rats were randomly divided into sham operation control group, model control group, evodiamine low(40 mg · kg-1) and high(80 mg· kg-1) dose groups. Evodiamine was intragastric administration for 14 consecutive days, and the sham operation or control rats were given with distilled water. After consecutive 14 d, the neointimal hyperplasia degree was observed by histopathological alterations and by calculating the proliferating cell nuclear antigen(PCNA) positive cells expression percentage in the common carotid arter-y. The levels of nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) in plasma of rats were measured respectively. The mRNA expressions of PCNA, endothelial nitric oxide synthase(eNOS) and SM α-actin in carotid artery wall were analyzed separately by real time RT-PCR. RESULTS: The neointimal hyperplasia was very serious, as evidenced by thickened neointima and narrowed lumen in carotid artery balloon-injured rats (model control group). Compared with the model control group, evodiamine 40 and 80 mg · kg-1 administration could significantly ameliorate the histopathology of common carotid artery, decrease the positive expression rate of PCNA, but increase the levels of NO, cGMP in plasma of rats, downregulate the PCNA mRNA expression and upregulate mRNA expressions of eNOS and SM α-actin, respectively. CONCLUSION: Evodiamine can depress the vascular neointimal hyperplasia induced by artery endothelia rubbing in rats, the mechanism may be related, at least partly, to the promotion of NO production.
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Objective To study the effects of protein kinase A (PKA) phosphorylation site' s mutation of mitofusin-2 (Mfn2) on intimal proliferation after balloon injury of carotid artery of rats. Method Rat model of carotid artery balloon injury was established and infected with Adv-LacZ, Adv-Mfn2, AdvMfn2-S442A or Adv-Mfn2-S442D from the peri-arterial sheathes of vessels, while phosphate buffered solution (PBS) used instead of above infectious adenovirus as uninfected group and sham operation as control group. The rats were sacrificed 14 days after balloon injury of carotid artery in order to measure the level of Mfn2 protein and the prol9iferation cell nuclear antigen (PCNA) with immunohistochemistry staining. The morphology of vessels was observed with HE staining. All data were statistically analyzed with ANOVA and Dunnett-t test. Results Fourteen days after surgery, the levels of Mfn2 protein significantly increased in arteries infected with Adv-Mfn2, Adv-Mfn2-S4442A and Adv-Mfn2-S442D compared with those in control group, sham operation group and Adv-LacZ infected group. The ratio of intimal area/medial area (I/M) and percentage of PCNA positive cells in both Adv-Mfn2 and Adv-Mfn2-S442A groups markedly decreased compared with control group (P ＜0. 01 ) . Compared with the Adv-Mfn2 group, the I/M and the percentage of PCNA positive cells reduced more significantly in Adv-S442A group (P ＜ 0. 01 ) . However,the I/M and the percentage of PCNA positive cells in Adv-LacZ and Adv-S442D groups were not significantly different from those found in the control group. Conclusions The over-expression of Mfn2 gene may effectively inhibit intimal proliferation after balloon injury of carotid artery of rats. The inhibitory effects of Mfn2-S442A are more obvious than those of Mfn2. However, the Mfn2-S442D is out of the inhibitory effect on neo-intimal proliferation.
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Objective To investigate the effect of Yiqi Huoxue prescriptions on prevention of neointima hyperplasia after carotid artery vascular injury in rats.Methods Ninty male SD rats were randomly divided into 6 groups:the model group,the aspirin group,the low-dose Yiqi Huoxue group,the mid-dose Yiqi Huoxue group and the high-dose Yiqi Huoxue group.All of the rats were fed with hypercholesterolemic diet for 30 days,then the 2.0F balloon catheter injury was perfomed through the right femoral artery into the left common carotid artery of rats,the balloon was inflated and deflated 3 times.The rats was given drugs from 1 d before injury and to 5 d,14 d,28 d after injury.Around 6 rats from each group were euthanized by exsanguination at appropriate time point.The injuried left common carotid arteries were carefully removed and harvested for HE staining.Results The neointima hyperplasia was faint at 5 d after the operation.Morphological analysis demonstrated that there were markedly differences in the area of neointima hyperplasia at 14 d or 28 d,the average thickness of neointima hyperplasia,the surplus area of lumen/the total area of lumen between the high-dose Yiqi Huoxue group and the model group(P
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Background The ubiquitin-proteasome pathway(UPP)involves 80%-90% degradation of all in- tracellular proteins.Both ubiquitin and rat component 3 of proteasome(RC3)are hence considered to be central me- diators for cell biology.Objective To evaluate the effect of simvastatin on neointimal hyperplasia and the expres- sion of ubiquitin and rat component 3 of proteasome(RC3)after balloon injury in carotid artery.Methods Thirty- two male SD rats were randomly to receive,low dose(0.4 mg/ng,n=8),or moderate(4 mg/ng,n=8),or high- dose(40 mg/ng,n=8)of simvastatin treatment for 28 days.Common carotid aortic artery was injuried rat ballon. The ratio of intima-media(I/M)thickness was determined.The expression level of ubiquitin and RC3 mRNA were assessed by RT-PCR.The expression level of ubiquitin protein was examined with immunohistochemistry. Results Simvastatin inhibited the expression of ubiquitin and RC3 mRNA and ubiquitin protein in dose dependent manner.The intima-media ratio(-50.2 %)and the expression of ubiquitin(-60.3 %)and RC3 mRNA and ubiq- uitin protein(-60.5 %)was reduced by the high dose simvastatin [40 mg/(kg?d)](P
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Objective:To observe the effect of Radix Paeoniae Rubra on intimal proliferation,activating of angiotensin Ⅱ(AgⅡ) and expression of Mitogen-activated protein kinase phosphatase-1(MKP-1) after carotid artery balloon injury in cholesterol-fed rabbits.Methods: Male rabbits were randomly divided into Radix Paeoniae Rubra(PPR) groups: high dose group(75、50、25g.kg-1.d-1;n=8),middle dose group(2g.kg-1.d-1),low dose group(1g.kg-1.d-1;n=8) and control group.Both groups received high fat forage(2% cholesterol + 5% lard).Balloon injury of carotid artery was performed.Carotid artery were harvested at the end of 10 weeks.Expression level of AgⅡwas measured by radioimmunoassay.MKP-1 expression was determined by RT-PCR.Immunohistochemical staining and morphological detection were adopted.Results: Compare with control group,expression of AgⅡ decreased obviously(P
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Objective:To observe the effect of Radix Paeoniae Bubra(RPB)on NADPH oxidase p22phox,monocyte chemotactic protein-1(MCP-1)mRNA expression and neointimal hyperplasia after carotid artery balloon injury in cholesterol-fed rabbits.Methods:The rabbit model of carotid balloon injury was established adopting Clowes method,and treated with extract of RPB.Component of neointima and expression of proliferating cell nuclear antigen(PCNA)and macrophage was determined by immunochemical stain.The collagen of typeⅠwas detected by special staining for blood vessels and the area of neointima was measured by image assay system.Expression of NADPH oxidase p22phox mRNA,MCP-1 mRNA was detected by in situ hybridization and transcription-polymerase chain reaction.Results:RPB can attenuate the neointima area and proliferation of collagen typeⅠinduced by balloon-injury,remarkable prevent the formation of restenosi,and down-regulate expression of NADPH oxidase p22 and MCP-1mRNA significantly and decrease the degree of macrophages infiltration especially in vessel wall of injuring carotid artery.Conclusions:RPB inhibited NADPH oxidase P22Phox and MCP-1 mRNA expression,and modestly reduced neointimal hyperplasia,which might be partly attributed to its antioxidant and inflammatory effects.
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Objective To investigate the role of interferon ?on smooth muscle cells proliferation and migration after balloon injury. Methods Animal model of rabbit iliac artery balloon injury was set up, smooth muscle cells derived from injured artery were cultured. Smooth muscle cells were divided into four groups ( control, TGF ? 1, IFN ?, TGF ? 1 and IFN ?).Cells from each group were treated with medium or TGF ? 1 (10 ng/ml) and/or IFN ?(500 u/ml) for 72 h separately. Smooth muscle cell proliferation was determined by cell count and MTT, migration of smooth muscle cells was also detected. Matrix metalloproteinase 2 was also detected by zymography. Results Our results showed that, compared with group control(2.875?0.323?10 5 cells/ml,279.9?8.129 ?m), TGF ? 1 increased cell count (4.188?0.239?10 5 cells/ml, P
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Objective To explore the expression of lectin-like oxidized low-density lipoprotein receptor-1 ( LOX-1) in a rat model after balloon arterial injury and investigate the effect of cilostazol on the mRNA and protein expressions of LOX-1. Methods Twenty-four male SPF SD rats were randomly divided into 3 groups ( n =8 for each group) ,sham operation group,balloon arterial injury group and cilostazol treatment group. For the rats from balloon arterial injury group and cilostazol treatment group,the left common carotid artery were in- juried by a balloon catheter. The expression of LOX-1 mRNA and protein in neointimal hyperplasia after balloon arterial injury were examined by RT-PCR and immunohistochemistry respectively. The contents of MDA in blood serum was examined by TBA. Results LOX-1 mRNA and protein were expressed in the neointimal hyperplasia after balloon arterial injury but not in the sham-operation group. The expression level of LOX-1 was significantly lower in the cilostazol treatment group than that in the model group ( P