ABSTRACT
Objective:To detect and verifica the gene profile difference of microRNA-146a (miR-146a) and its role in the pro-liferation of vascular smooth muscle cells (VSMCs) by gene chip technology. Methods: Artificially synthesized miR-146a mimics(50 nmol/L) ,miR-146 inhibitor ( 50 nmol/L ) , scramble ( 50 nmol/L ) and PBS were transfected into cultured primary rat VSMCs in vitro. After transfection,Real time PCR was used to measure the levels of miR-146a and the cell counting kit 8(CCK8) was employed to investigate the proliferation of VSMCs. The VSMCs interfered by miR-146a inhibitor or miR-146a control were examined by gene chips and the profile of gene were analyzed by bioinformatics technology to detect the different genes and signal transduction pathway. The changes in mRNAs and proteins were accessed separately by Real time PCR and Western blot. Results: Compared with sham and control VSMCs,miR-146a expression level was significantly decreased in treatment with miR-146a inhibitor(P0. 05),however,the mRNA and protein expression levels of cyclin D1 significantly increased in treatment with miR-146a mimics VSMCs group and decreased in miR-146a inhibitor VSMCs group ( compared with sham VSMCs group, both P<0. 05 ) . Conclusion: Our data indicated that miR-146a may promote the proliferation of rat VSMCs by up-regulating cyclin D1 expression.
ABSTRACT
<p><b>OBJECTIVE</b>To assess the relationship between visit-to-visit blood pressure (BP) variability (BPV) and vascular endothelial function in a cohort of elderly hypertensive patients.</p><p><b>METHODS</b>A total of 174 elderly patients with essential hypertension were included in the study. The participants had their office BP measured during the 12-month follow-up. Right brachial artery diameter was assessed at rest, during reactive hyperemia (flow-mediated dilation, FMD), and after nitroglycerin administration (nitroglycerin-mediated dilation, NMD). The participants were divided into two groups according to FMD% or FMD/NMD ratio. The correlations between BPV and endothelial function were analyzed by univariate analysis and multiple linear regression analysis.</p><p><b>RESULTS</b>The participants classified as having a decreased endothelial function according to FMD/NMD ratio had significantly lower FMD% and higher BPV and NMD% (P<0.05). The percentage of CCBs use in normal endothelial function group was significantly higher than that in decreased endothelial function group (79.55% vs 63.95%, P<0.05). Multiple linear regression analysis revealed a significant negative association between FMD/NMD ratio and BPV, and this association remained significant after adjustment for age, body mass index, and mean BP levels.</p><p><b>CONCLUSIONS</b>FMD/NMD ratio is a better marker of endothelial function than FMD, and an increased visit-to-visit variability of BP is associated with a decreased endothelial function.</p>
Subject(s)
Aged , Humans , Blood Pressure , Brachial Artery , Endothelium, Vascular , Essential Hypertension , Hypertension , Multivariate Analysis , Nitroglycerin , Regression Analysis , VasodilationABSTRACT
BACKGROUND:Skeletal muscle satelite cels are muscle-derived stem cels with proliferation and differentiation potential distributing between the muscle cel membrane and the base film. Studies have shown that skeletal muscle satelite cels are of efficacy and safety, but the survival rate of the transplanted stem cels is very low, which greatly limits the application of skeletal muscle satelite cels. OBJECTIVE: To observe the effects of Fasudil on apoptosis of skeletal muscle satelite cels induced by H2O2. METHODS: Skeletal muscle satelite cels cultured in vitro were randomly divided into three groups including H2O2group, H2O2+Fasudil group (Fasudil group) and control group. Apoptosis rates were observed by flow cytometry. The concentrations of interleukin-4 and tumor necrosis factor-a in each group were detected by ELISA. Western blot was employed to measure the protein level of Bax in each group. RESULTS AND CONCLUSION: Compared with the H2O2group, a significant decrease was found in the apoptosis rate of cels, protein level of Bax, and concentrations of interleukin-4 and tumor necrosis factor-a in the Fasudil group (alP < 0.05). These findings indicate that Fasudil can play anti-apoptosis protection by inhibiting Rho-kinase signaling pathway, which may be related to the reduced expression of Bax.
ABSTRACT
AIM:To investigate the proliferation property of stable chemerin gene knockdown vascular smooth muscle cells ( VSMCs) and to explore its mechanism .METHODS:The normal VSMCs , chemerin gene interfering control VSMCs and stable chemerin gene knockdown VSMCs were divided into normal group , PDGF group, control group and knockdown group .The VSMCs in PDGF group were given platelet-derived growth factor-BB ( PDGF-BB) to initiate proli-feration.The cell counting and BrdU assay were employed to investigate the proliferation property of VSMCs .The mitogen-activated protein kinase ( MAPK) signal pathway was determined by Western blot .RESULTS:The cell number and BrdU A value in PDGF-BB-treated VSMCs significantly increased as compared with the normal VSMCs ( P<0.05 ) .Compared with the normal VSMCs , the chemerin knockdown VSMCs showed obviously decreased cell number and BrdU A value ( P<0.05).Simultaneously, no significant difference in the proliferation of VSMCs between the normal VSMCs and the control VSMCs was observed.No significant difference of the protein levels of p-ERK1/2, ERK1/2, p-p38 MAPK and p38 MAPK among 4 kinds of VSMCs was found .The protein level of p-JNK in PDGF-BB-treated VSMCs was up-regulated, while it was down-regulated in chemerin knockdown VSMCs compared with the normal VSMCs .CONCLUSION: Chemerin pro-motes the proliferation of mouse vascular smooth muscle cells by up-regulating p-JNK production .
ABSTRACT
BACKGROUND:Previous studies have shown that a certain dose of acidic fibroblast growth factor can promote skeletal muscle satelite cel proliferationin vitro. OBJECTIVE:To investigate the effects of transfection with acidic fibroblast growth factor by electroporation on growth, proliferation and differentiation of skeletal muscle satelite cels. METHODS: Skeletal muscle satelite cels were cultured and purified, and then transfected with plasmid pSectag-GFP-aFGF by electroporation. The expression of green fluorescent protein was observed under fluorescence microscope, and the transfection efficiency was calculated. After transfection, cel cycle was analyzed by flow cytometry to draw the growth curve of skeletal muscle satelite cels. Western blot assay was employed to measure protein level of acidic fibroblast growth factor. RESULTS AND CONCLUSION: (1) Immunocytochemistry detection: The skeletal muscle satelite cels were positive for a-sarcomeric actin. (2) Transfection efficiency: At 12 hours after transfection with pSectag-aFGF, several cels showed green fluorescence, and the green fluorescent expression reached the peak at 72-96 hours after transfection with a positive rate of about 90%. (3) Cel cycle: After electrotransfection, the proportion of cels at S phase in the electroporation group was higher than that in the control group (P < 0.05). (4) Cel growth curve: At 3 days after electrotransfection, the cels entered logarithmic growth phase but the proliferation slowed down at 5 days. (5) Differentiation capacity: There were fewer myotubes and aging cels in the electroporation group than the control group. (6) Western blot assay: Acidic fibroblast growth factor protein was highly expressed in the cels transfected with target gene detected by western blot assay. These findings indicate that by using electroporation method, acidic fibroblast growth factor can be transferred into skeletal muscle satelite cels and have a high-efficiency and long-term expression, which can promote the proliferation of skeletal muscle satelite cels and inhibit formation of myotubes.
ABSTRACT
<p><b>OBJECTIVE</b>[corrected] To understand the role of miRNA-146a in the proliferation and migration of primarily cultured rat vascular smooth muscle cells (VSMCs) and investigate the mechanisms.</p><p><b>METHODS</b>Primarily cultured rat VSMCs were transfected with a synthesized miRNA-146 inhibitor, a scramble sequence or PBS via Lipofectamine2000. Cell counting kit 8 (CCK8) and transwell assay were employed to assess the proliferation and migration of the transfected cells, and the expressions of nuclear factor-κB p65 (NF-κBp65) and proliferation cell nuclear antigen (PCNA) were detected using Western blotting.</p><p><b>RESULTS</b>A 48-h transfection of the VSMCs with miRNA-146 inhibitor caused significantly lowered miRNA-146a expression as compared with that in VSMCs transfected with the scramble sequence or PBS (P<0.01), resulting also in lowered proliferative and migration ability of the cells (P<0.01). The expression levels of NF-κBp65 and PCNA were remarkably lower in cells transfected with miRNA-146 inhibitor than in the cells in the other two groups (P<0.05).</p><p><b>CONCLUSION</b>miRNA-146a is capable of promoting the proliferation and migration of rat VMSCs probably by enhancing the expression of NF-κBp65.</p>
Subject(s)
Animals , Male , Rats , Cell Movement , Cell Proliferation , MicroRNAs , Genetics , Muscle, Smooth, Vascular , Cell Biology , Metabolism , Myocytes, Smooth Muscle , Metabolism , Primary Cell Culture , Proliferating Cell Nuclear Antigen , Genetics , Metabolism , Rats, Sprague-Dawley , Transcription Factor RelA , Genetics , Metabolism , TransfectionABSTRACT
AIM: To observe the effects of H_2O_2 on apoptosis of skeletal muscle satellite cells (SMSC)and mitochondrial membrane potential (MMP), and protective effect of erythropoietin(EPO). METHODS: SMSC in vitro were divided into three groups: H_2O_2 group, H_2O_2+EPO group and control. Apoptosis rate and the means were obverted by monofluorescence flow cytometry. The morphological change of apoptosis cells were observed under fluorescence microscopy after Hoechst 33258 staining. RESULTS: The cells in H_2O_2 group show the highest apoptosis rate (22.13±1.79)%. In H_2O_2+EPO group, apoptosis rate were (16.47±2.53)%, (4.97±0.55)% and (2.93±0.47)% according to the EPO treated levels (10, 20 or 40 kU/L), respectively. MMP level in H_2O_2 group was the lowest 9.70±0.09. MMP levels in H_2O_2+EPO group were 12.67±0.32, 27.90±0.66, 44.53±0.93, respectively according to the EPO treated levels (10, 20 or 40 kU/L). In control group, apoptosis rate was 1.93±0.57 and MMP was 51.37±0.64. In H_2O_2 group and H_2O_2+ low dosage EPO group, Hoechst 33258 staining showed obvious apoptosis. CONCLUSION: EPO inhibits the apoptosis induced by H_2O_2 and stabilizes the MMP, which is related to the dosage of EPO.
ABSTRACT
BACKGROUND:Restenosis and lumina loss limit further application of balloon extension and stent implantation.Effect of tunica intima proliferation and apoptosis in restenosis and the intervention method are exploring.OBJECTIVE:To investigate the influence of Rosuvastatin on the vascular smooth muscle cells proliferation and apoptosis in rats with carotid artery injury established by Medtronic balloon.METHODS:The male SD rats were randomly and equally divided into injury group and treatment group.Each rat was subjected to balloon injury on the lift common carotid artery,and control artery without balloon injury on the right artery served as control group.Treatment group rats were given Rosuvastatin(dissolved in Nacl)5 mg/kg per day 3 days before injury,while the injury group rats were given 9 g/L NaCl.At 7 and 14 days after injury,the common carotid arteries were harvested for HE staining.SM α-actin and proliferating celI nuclear antigen were detected by immunohistochemistry.In addition,smooth muscle cells apoptosis was detected by TUNEL.RESULTS AND CONCLUSION:The neointimal area and the area ratio of neointimal/media were decreased in treatment grouP significantly at 14 days compared with injury group(P<0.05),and neointimal area increased by 26%:positive cell rate of proliferating cell nuclear antigen was decreased,but apoptosis cells were increased cornpared with the injury group(P<0.05).Results showed that Rosuvastatin prior to balloon injury inhibited neointimal proliferation and neointimal celI proliferation following balloon Injury,promoted smooth muscle cells apoptosis,ultimately reducing neointimaI formation and inhibiting restenosis.
ABSTRACT
Objective To investigate the effect of hydrogen peroxide (H_2O_2) on apoptosis and calcium ion concentration of skeletal muscle satellite cells (SMSCs) in rats, and to explore the protective effect of erythropoietin (EPO).Methods The cultured SMSCs were divided into five groups: control group,H_2O_2 group, 10, 20 and 40 U/ml EPO intervention groups.Apoptosis rates and calcium ion concentration of SMSCs were analyzed by flow cytometry, and the morphology of apoptotic cells was observed by Hoechst33258 staining.Results The apoptosis rates showed significant differences (all P<0.05) among (1.93±0.57)% in control group, (22.13±1.79)% in H_2O_2 group, (16.47±2.53)%, (4.97±0.55)% and (2.93±0.47)% in 10, 20 and 40 U/ml EPO intervention groups, respectively.And calcium ion concentrations in SMSCs were 12.67 ±0.32, 27.90±0.06 and 44.53±0.93 in 10, 20 and 40 U/ml EPO intervention groups, respectively.There was significant difference in calcium ion concentration between H_2O_2 group and control group (9.70±0.09 vs.51.37± 0.64, P< 0.05).Morphology of apoptosis was observed by Hoeehst33258 dye stains in 10, 20 U/ml EPO intervention group and H_2O_2 group, while there were less apoptotic bodies in 40 U/ml EPO intervention group and control group.Conclusions EPO might have protective effects on SMSCs injured by H_2O_2 through inhibiting apoptosis and calcium ion releasing from SMSCs.
ABSTRACT
BACKGROUND: Inflammation plays an important role in vessel proliferation after balloon injury. Reducing inflammatory reaction may lighten the ocurrence and development of the restenosis after angioplasty. Studies have demonstrated that PPAR_Y excitomotor has inhibitory effects on inflammation development. OBJECTIVE: To observe the changes in inflammatory factors after carotid artery balloon injury in rats and the intervention of PPARy excitomotor rosiglitazone. DESIGN, TIME AND SETTING: The randomized, controlled animal experiment was performed at the Central Laboratory of Shenzhen People's Hospital from January to June 2009. MATERIALS: Male SPF SD rats weighing about 350 g were selected to generate models of carotid balloon injury. METHODS: SD rats were equally and randomly divided into 3 groups: the control group, the balloon injury group and the rosiglitazone group. The left common carotid arteries were injured by balloon in the balloon injury group and the rosiglitazone group. The control group received sham operation. The rosiglitazone group was administered rosiglitazone daily by gavage,which began 4 days before operation and continued until harvesting.Accordingly,the control group and the balloon injury group were administered normal saline daily by gavage. MAIN OUTCOME MEASURES: All rats were executed under anesthesia at 14 days after operation, respectively to harvest left common carotid artery samples. The vessels were stained by hematoxylin-eosin, and Neointimal area (NIA) and media area (MA) as well as NIA/MA were calculated. Real time RT-PCR and Western Blot method were used to assay the expression of interleukin (IL)-6, IL-10, IL-17A mRNA and the distribution of nuclear factor (NF)-kB protein. expression levels of IL-6 and IL-17A mRNA in the rosiglitazone group were significantly lower than the balloon injury group, but higher than the Control group( P < 0.05), The expression levels of IL-10 mRNA in the rosiglitazone group were higher than the the rosiglitazone group was down-regulated, and lower than the balloon injury group, but higher than control group (P < 0.05). CONCLUSION: Rosiglitazone can regulate the expression of II-6 IL-10 IL-17A mRNA and the balance of inflammatory factors via NF-kB,inhibit the inflammatory reaction of injured vessels and may contribute to lighten the restenosis of injured vessels.
ABSTRACT
BACKGROUND:Inflammation plays an important role in vessel proliferation after balloon injury.Reducing inflammatory reaction may lighten the ocurrence and development of the restenosis after angioplasty.Studies have demonstrated that PPAR? excitomotor has inhibitory effects on inflammation development.OBJECTIVE:To observe the changes in inflammatory factors after carotid artery balloon injury in rats and the intervention of PPAR? excitomotor rosiglitazone.DESIGN,TIME AND SETTING:The randomized,controlled animal experiment was performed at the Central Laboratory of Shenzhen People's Hospital from January to June 2009.MATERIALS:Male SPF SD rats weighing about 350 g were selected to generate models of carotid balloon injury.METHODS:SD rats were equally and randomly divided into 3 groups:the control group,the balloon injury group and the rosiglitazone group.The left common carotid arteries were injured by balloon in the balloon injury group and the rosiglitazone group.The control group received sham operation.The rosiglitazone group was administered rosiglitazone daily by gavage,which began 4 days before operation and continued until harvesting.Accordingly,the control group and the balloon injury group were administered normal saline daily by gavage.MAIN OUTCOME MEASURES:All rats were executed under anesthesia at 14 days after operation,respectively to harvest left common carotid artery samples.The vessels were stained by hematoxylin-eosin,and Neointimal area(NIA) and media area(MA) as well as NIA/MA were calculated.Real time RT-PCR and Western Blot method were used to assay the expression of interleukin(IL)-6,IL-10,IL-17A mRNA and the distribution of nuclear factor(NF)-?B protein.RESULTS:Of the 36 rats,5 were excluded due to failed modeling or death,and 31 rats were included in final analysis.①The expression levels of IL-6 and IL-17A mRNA in the rosiglitazone group were significantly lower than the balloon injury group,but higher than the control group(P
ABSTRACT
Objective To assess the safety and efficiency of Angioseal device in patients undergoing coronary percutaneous procedure.Methods A prospective trial was carried out in 260 patients undergoing angiograph and angioplasty during october 2002 to July 2003.All patients were divided into two groups: Angioseal closure device and manual compression.Results In angiography,the time to hemostasis was (1.8?0.9)min by Angioseal and (25.3?13.4)min by manual compression(P
ABSTRACT
<p><b>OBJECTIVE</b>To assess the relationship between myocardial regional perfusion using second harmonic myocardial contrast echocardiography (MCE) by venous injection of Levovist and coronary artery stenosis detected by coronary angiography to determine whe ther MCE can be used to detect coronary artery disease (CAD) and its sensitivity and specificity for detecting CAD.</p><p><b>METHODS</b>Thirty-six patients who underwent coronary artery angiography and MCE formed the study groups. Ten myocardial segments (5 each in the apical two- and four-chamber views) from the images were scored for detecting myocardial perfusion as follows: 1, normal perfusion; 2, decreased perfusion; and 3, perfusion defect. The arteries were classified as normal or diseased. The diseased arteries were classified into three groups according to the perfusion scores.</p><p><b>RESULTS</b>There were significant differences in coronary diameter stenosis among the different perfusion score groups (P < 0.001). There were 10 total occluded arteries, and the myocardial perfusion scores were different because of different collateral circulation. In the normal perfusion group (Group A), the coronary diameter stenosis was 65% +/- 12%, and the myocardial perfusion score index was 1 +/- 0.00. In the decreased perfusion group (Group B), the average coronary diameter stenosis was 82% +/- 8%, and the myocardial perfusion score was 1.93 +/- 0.16. The diameter stenosis was less than 85% in 63% of the coronary arteries (including diameter stenosis < or = 75% in 12% of the vessels). The diameter stenosis was 85%-90% in 22% of the coronary arteries and > 90% in 15% of the arteries. In the perfusion defect group (Group C), the average diameter stenosis was 90% +/- 6%, and the myocardial perfusion score index was 2.89 +/- 0.24. The diameter stenosis was > or = 85% in 94% of the coronary arteries, and the diameter stenosis was < 85% and > 75% only in 6% of the coronary arteries. The overall sensitivity and specificity of MCE in identifying angiographic coronary diameter stenosis was 67% and 100%, respectively. The false negative rate was 32.6% for the 108 coronary arteries. Further subdivided analysis showed the sensitivities in Groups A, B and C were 0, 100%, and 100%, respectively. The sensitivity increased with increased lumen diameter stenosis of coronary arteries.</p><p><b>CONCLUSIONS</b>There is a close relationship between coronary artery stenosis and MCE perfusion scores. MCE with venous injection of new generation contrast can define the presence of CAD and lesion graded classifications. Some disagreements between perfusion score and coronary diameter of stenosis may indicate other factors such as different collateral circulation, which should be further investigated. As artery stenosis increases, the sensitivity of MCE is increased.</p>
Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Coronary Circulation , Coronary Disease , Diagnostic Imaging , Echocardiography , Sensitivity and SpecificityABSTRACT
Objective To evaluate the vascular remodeling features in the elderly hypertensives through detecting the changes of large arterial structure and function by ultrasonography. Methods High resolution ultrasound was applied to examine 52 lightly or moderate elderly hypertensives(LMEH), 52 elderly normal subjects(EN) and 25 adult normal subjects(AN). Carotid and humeral arterial intimal-medial thickness (IMT), lumen diameter(r) and IMT/r were determined as the index for the changes of arterial structure, and the humeral arterial endothelium-dependent vasodilation and cross-sectional compliance were used to reflect the changes of arterial functions. Results Carotid IMT and lumen diameter were larger in LMEH group than those in EN group(0.97?0.21)% vs. (0.82?0.12)%, (7.41?2.13)% vs. (6.38?0.45)%, all P
ABSTRACT
Objective To evaluate the safety and feasibility of the implication of Cypher~(TM) drug-eluting stent in primary PTCA of acute myocardial infarction.Methods From November,2002 to November,2004,77cases of acute myocardial infarction patients enrolled into our hospital were treated by PCI within 12 hours after the heart attack.All patients were divided into two groups:Cypher~(TM) drug-eluting stent group(38) and conventional stainless steel stent group(39).Results The success rate of PCI were 100% in either group.38 Cypher~(TM) drug-eluting stent and 39 conventional stainless steel stent were implanted respectively.There was no significant difference in CAG result or clinical state among these two groups.Conclusion Cypher~(TM) drug-eluting stent can be safely and efficiently used in patients with acute myocardial infarction.
ABSTRACT
Objective To study the results from Cutting balloon for stiff and uncompliant coronary lesion. Methods Thirty-three patients with coronary artery disease underwent conventional coronaryangioplasty with high pressure balloon. Cutting balloon was used for those lesions after they failed in high pressure balloon angioplasty. Results The total success rate for cutting balloon for stiff and uncompliant lesion was 87.9%. We got remarkable results from cutting balloon (MLD 2.47?0.54 mm; DS 16%?15%) with less dilation pressure and dilation time (8.93?1.23 atm; 1.65?0.44 min). Conclusion Our results show that cutting balloon can be used in the stiff lesion and some calcified lesion which was once considered to be contraindiction it could be a high procedural successful rate and very safe with less complications compared with other kinds of new technology.
ABSTRACT
Objective To study the puncture method of Angioseal closure device by femoral artery. Methods A prospective trial was carried out in 80 patients using Angioseal closure device in angiography and angioplasty. All patients were divided into tow groups, according to the horizontal distance between the entrance of skin and the entrance of the femoral artery: Group A, the distance 1.5 cm. Results 1. The rate of successful puncture was 92% in group A and 81% in group B (P