Efecto del Camphenol Plus sobre el músculo liso vascular de vena porta en ratas Wistar / Effect of Camphenol Plus on portal vein vascular smooth muscle in Wistar rats

RESUMEN Introducción: uno de los antisépticos comúnmente empleado en Estomatología desde el pasado siglo y que mantiene su uso hasta la actualidad, lo constituye el Camphenol Plus. Son escasos los reportes científicos de su efecto sobre el endotelio y la dinámica contráctil del músculo liso vascular, en especial de tejidos venosos como la vena porta hepática. Objetivo: determinar el efecto del Camphenol Plus sobre el músculo liso vascular de la vena porta. Métodos: se realizó una investigación experimental preclínica, con la utilización de 21 venas porta obtenidas de ratas Wistar. Las preparaciones realizadas se colocaron en baño de órganos, se registró la tensión desarrollada por el músculo liso vascular tras la adición de diez microlitros de Camphenol Plus, en diferentes concentraciones y durante diferentes intervalos de tiempo. Resultados: el Camphenol Plus, tras la preactivación del musculo liso vascular de la vena porta, indujo vasorelajación, la que se incrementó durante todo el tiempo de estudio y según el incremento de las concentraciones del medicamento. Existieron diferencias significativas entre los valores de tensión promedios registrados en los diferentes intervalos de tiempo con los de la tensión espontánea basal y la tensión base inicial. Conclusiones: el Camphenol Plus, indujo "in vitro", relajación de la musculatura lisa de la vena porta a través de un acoplamiento excitación-contracción de tipo farmacomecánico.

ABSTRACT Introduction: Camphenol Plus is one of the antiseptics commonly used in Dentistry since the last century and still in use today. There are few scientific reports of its effect on the endothelium and contractile dynamics of vascular smooth muscle, especially in venous tissues such as the hepatic portal vein. Objective: to determine the effect of Camphenol Plus on the vascular smooth muscle of the portal vein. Methods: a preclinical experimental investigation was carried out using 21 portal veins obtained from Wistar rats. The preparations were placed in an organ bath and the tension developed by the vascular smooth muscle was recorded after the addition of ten microliter of Camphenol Plus, at different concentrations and during different time intervals. Results: Camphenol Plus, after the preactivation of the vascular smooth muscle of the portal vein, induced relaxation, which increased throughout the study time and according to the increase in drug concentrations. There were significant differences between the average tension values recorded in the different time intervals with those of the basal spontaneous tension and the initial baseline tension. Conclusions: Camphenol Plus induced "in vitro" relaxation of portal venous smooth muscles through a pharmacomechanical excitation-contraction coupling.

Mechanisms for Nogo-B mediating mitochondrial reactive oxygen species generation during vascular remodeling in mice / 中华老年医学杂志

Objective:To investigate the role of Nogo-B in mitochondrial reactive oxygen species(m-ROS)production and vascular remodeling in vascular smooth muscle cells(VSMCs), and to further clarify the molecular mechanism for Nogo-B in m-ROS production via regulating ATP synthase expression.Methods:A mouse model of vascular remodeling was established.The expression of Nogo-B in mouse smooth muscle cells was detected.Forty mice were divided into the AAV-NC+ control, AAV-NC+ angiotensin Ⅱ(AngⅡ), AAV-Nogo-B+ control and AAV-Nogo-B+ AngⅡ groups(n=10 for each group). VSMCs cultured in vitro were divided into the control group and the Nogo-B overexpression group(Ad-Nogo-B). The expression of Nogo-B in VSMCs in vitro stimulated with AngⅡ was detected.Through experiments conducted in vivo, Nogo-B was overexpressed in VSMCs, then VSMCs were stimulated with AngⅡ, and m-ROS production, tissue fibrosis and mitochondrial function were examined.The regulatory effects of Nogo-B on m-ROS production were explored.Molecular mechanisms for NoGO-B in vascular remodeling via regulating ATP synthase/m-ROS production were examined. Results:Compared with the control group, the expression of Nogo-B was decreased in VSMCs treated with AngⅡ(0.36±0.13 vs.1.00±0.13, t=8.44, P<0.05). The production of m-ROS, fibrosis and mitochondrial dysfunction were increased in VSMCs during vascular remodeling( P<0.05), while overexpression of Nogo-B significantly reduced m-ROS production, fibrosis and mitochondrial dysfunction( P<0.05). ATP synthase expression in VSMCs was positively regulated by Nogo-B.ATP synthase expression was higher in the AAV-Nogo-B+ AngⅡ group than in the AAV-NC+ AngⅡ group(0.86±0.14 vs.0.49±0.17, t=-3.97, P<0.05). The production of m-ROS was reduced by Nogo-B and was lower in the AAV-Nogo-B+ AngⅡ group than in the AAV-NC+ AngⅡ group(1.28±0.34 vs.3.26±0.57, t=7.18, P<0.05). Meanwhile, the ability of Nogo-B to mitigate the deleterious effects of oxidative stress in VSMCs could be offset by oligomycin. Conclusions:Nogo-B participates in vascular remodeling by regulating ATP synthase-mediated m-ROS production.

Diagnostic accuracy of retrospective application of the Vesical Imaging-Reporting and Data System: preliminary results / Resultados preliminares da aplicação retrospectiva do Vesical Imaging-Reporting and Data System

Abstract Objective: To evaluate the retrospective accuracy of the Vesical Imaging-Reporting and Data System (VI-RADS) in detecting muscle invasion in bladder cancer. Materials and Methods: We investigated patients who underwent pelvic magnetic resonance imaging and were submitted to transurethral resection of a bladder tumor between 2015 and 2018. Thirty cases were reviewed by radiologists blinded to the final clinical stage. The VI-RADS score was applied and compared with the histopathological findings in the surgical specimen. Results: Of the 30 patients with suspicious bladder lesions, 5 (16.6%) had benign histopathological findings, 17 (56.6%) had non-muscle-invasive bladder cancer, and 8 (26.6%) had muscle-invasive bladder cancer. The optimal criterion to detect muscle-invasive bladder cancer was a final VI-RADS score > 3, for which the sensitivity and specificity were 100% (95% CI: 56.0-100%) and 90.9% (95% CI: 69.3-98.4%), respectively. Conclusion: The VI-RADS appears to estimate correctly the degree of muscle invasion in suspicious bladder lesions. However, prospective studies evaluating larger samples are needed in order to validate the method.

Resumo Objetivo: O objetivo deste estudo foi avaliar retrospectivamente a acurácia do Vesical Imaging-Reporting and Data System (VI-RADS) para detectar invasão muscular em câncer de bexiga. Materiais e Métodos: Foram inseridos pacientes submetidos a ressonância magnética pélvica e a ressecção transuretral de bexiga entre 2015 e 2018. Trinta casos foram revisados, sem o conhecimento do estágio clínico final. O escore do VI-RADS foi aplicado e comparado aos achados histopatológicos da ressecção transuretral de bexiga. Resultados: Entre os 30 pacientes com lesões vesicais suspeitas, 5 (16,6%) tinham achados histopatológicos benignos, 17 (56,6%) tinham câncer de bexiga não músculo invasivo e 8 (26,6%) tinham câncer de bexiga músculo invasor. O critério ideal para detectar câncer de bexiga músculo invasor foi o escore final do VI-RADS > 3, em que sensibilidade e especificidade foram, respectivamente, 100% (IC 95%: 56,0-100%) e 90,9% (IC 95%: 69,3-98,4%). Conclusão: O VI-RADS parece estimar corretamente o grau de invasão muscular em lesões suspeitas da bexiga; no entanto, estudos maiores e prospectivos são necessários para validar o método.

Role of DNA Methylation in the Development and Differentiation of Intestinal Epithelial Cells and Smooth Muscle Cells

The mammalian intestine contains many different cell types but is comprised of 2 main cell types: epithelial cells and smooth muscle cells. Recent in vivo and in vitro evidence has revealed that various alterations to the DNA methylation apparatus within both of these cell types can result in a variety of cellular phenotypes including modified differentiation status, apoptosis, and uncontrolled growth. Methyl groups added to cytosines in regulatory genomic regions typically act to repress associated gene transcription. Aberrant DNA methylation patterns are often found in cells with abnormal growth/differentiation patterns, including those cells involved in burdensome intestinal pathologies including inflammatory bowel diseases and intestinal pseudo-obstructions. The altered methylation patterns being observed in various cell cultures and DNA methyltransferase knockout models indicate an influential connection between DNA methylation and gastrointestinal cells' development and their response to environmental signaling. As these modified DNA methylation levels are found in a number of pathological gastrointestinal conditions, further investigations into uncovering the causative nature, and controlled regulation, of this epigenetic modification is of great interest.

Effects of ezetimibe and rosuvastatin on the expression of Caveolin-1 mRNA in smooth muscle derived foam cells / 中国医师杂志

Objective To investigate the influence of ezetimibe combined with rosuvastatin on expression of Caveolin-1 in smooth muscle derived foam cells induced by oxidized low density lipoprotein (oxLDL).Methods The rat thoracic aortic smooth muscle cells (VSMCs) were selected from generations 3-5 in logarithmic growth cycle.The rat vascular smooth cells were induced using oxidized low density lipoprotein(ox-LDL 50 μg/ml for 48 h) to establish foam cell model.The normal cultured rat thoracic aortic smooth muscle cells were used as blank control group.Foam cells were divided into foam cell group,different concentrations of ezetimibe group,different concentrations of rosuvastatin group,combination group.The foam cells were incubated with different doses of ezetimibe (3.0,10.0,30.0 μmol/L) or rosuvastatin (0.1,1.0,5.0 μmol/L) for 24 h,or cultured with rosuvastatin 5.0 μmol/L + ezetimibe 30.0 μmol/L in combination groups.Oil red O staining was used to identify foam cell models.The expression of Caveolin-1 mRNA was detected by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR).Results Compared with blank control group,the mRNA expression of Caveolin-1 in foam cell group were decreased significantly [(0.248 7 ± 0.042 0) vs (1.004 1 ± 0.017 1),P ＜ 0.05].Compared with foam cell group,the mRNA expression of Caveolin-1 was increased in a dose-dependent manner in ezetimibe group and rosuvastatin group [(0.371 3 ±0.025 2),(0.489 8 ±0.027 9),(0.726 1 ±0.029 1) vs (0.248 7 ±0.042 0);(0.460 2±0.022 8),(0.623 7 ±0.028 8),(0.751 8 ±0.043 1) vs (0.248 7 ±0.042 0),P ＜0.05].Compared with the ezetimibe (30.0 μmol/L) and the rosuvastatin (5.0 μmol/L),the mRNA expression of Caveolin-1 in combined group were increased,the difference was statistically significant [(0.726 1 ±0.029 1),(0.751 8 ± 0.043 1) vs (0.937 6 ± 0.029 7),P ＜ 0.05].Conclusions Ezetimibe and rosuvastatin can promote the reverse transport of cholesterol (RCT) in smooth muscle derived foam cells by upregulating expression of Caveolin-1 mRNA.And the combination of ezetimibe (30.0 μmol/L) and rosuvastatin (5.0 μmol/L) has more significant effect.

Inhibitory effects of solid lipid nanoparticles of rhynchophylline on proliferation of rat vasculars mooth muscle cells induced by TGF-β1 / 国际中医中药杂志

Objective To observe the effect of Rhy-SLN on the proliferation of rat vascular smooth muscle cells (VSMC) induced by TGF-β1, and explore the mechanism. Methods The primary culture of rat thoracic aortic vascular smooth muscle cells was studied by tissue block culture method. The cells were divided into the control group, TGF-β1 group, TGF-β1+ the high, medium and low dosage groups of Rhy-SLN. In addition to the control group, the cells of the other groups were involved in the intervention of TGF-β1 of 20 g/L, and the high, medium and low dosage groups of Rhy-SLN cells were involved in the intervention of 25, 50, 100 mg/L of the hook teng solid lipid nanoparticles. After 24 hours of culture, MTT assay was used to detect cell proliferation inhibition rate in each group, and the cell cycle was detected by flow cytometry. The expression of c-myc and c-Fos protein in each group was detected by Western blot method. Results Compared with the TGF-β1 group, the absorbance value (0.457 ± 0.046 vs. 0.975 ± 0.049) of TGF-β1+ rhy-sln high dose group significantly decreased (P<0.01); the number of S phase cells (15.87% ± 2.47%, 15.23% ± 1.69%, 17.02% ± 2.87% vs.38.58% ± 2.68%)of TGF-β1+rhy-sln in each dose group significantly decreased(P<0.01);The c-myc(48.65 ±3.65,50.69 ± 4.16,55.29 ± 3.67 vs.68.21 ± 3.25)and c-Fos(38.78 ± 4.25,43.56 ± 3.69,46.58 ± 3.57 vs.66.54 ± 4.09) of the TGF-β1+ rhy-sln each dose group significantly decreased (P<0.01). Conclusions The Rhy-SLN can inhibit the proliferation of VSMC in rats induced by TGF-β1.Its mechanism is related to the conversion of G0/G1 phase to the S phase and the expression of the reduction of c-myc and c-fos protein.

The protective mechanism of D-pinitol on advanced glycation end products-induced proliferation and migration in vascular smooth muscle cells / 中华老年医学杂志

Objective To investigate the effect and its mechanism of D-pinitol on advanced glycation end products(AGEs)-induced proliferation and migration in mouse aortic vascular smooth muscle cells (VSMC).Methods VSMCs were isolated from mouse aorta and cultured in vitro.Effects of different concentrations of D-pinitol on proliferation and migration of VSMCs were observed by using the AGEs-induced glycosylation injury model of VSMCs.Cell proliferation and migration were detected by CCK-8 assay and cell scratch,respectively.The protein expression levels of transforming growth factor-β1(TGF-β1),p-smad2,p-smad3 and asporin were determined by Western blot.Results Compared with control group,AGEs group showed the increased protein expression levels of asporin,TGF-β1,p-smad2 and p-smad3 (40.06 ± 4.50 vs.17.47 ± 0.57),(55.25 ± 2.07 vs.14.42± 2.07),(0.97 ± 0.02 vs.0.47 ± 0.02),(0.45±0.01 vs.0.26 ± 0.02),all P＜ 0.01.Compared to AGEs group,D-pinitol group could inhibit the cell proliferation and migration and cause dose-dependent decreases of protein expressions of TGF-β1,p-smad2,p-smad3 and asporin(P ＜ 0.05 or P＜0.01).Conclusions D-pinitol can inhibit AGEs-induced cell proliferation and migration in mouse aortic VSMCs.Asporin may participate in the VSMCs extracellular matrix remodeling via TGF-β/smad pathway.

Research advances in signal pathway and molecular mechanism of vascular calcification / 中华老年医学杂志

Vascular disease,such as atherosclerosis and diabetic vasculopathy,is frequently complicated by vascular calcification.Previously believed to be an end-stage process of unregulated mineral precipitation,it is now well established to be a multi-faceted disease.The numerous regulatory signaling pathways that influence vascular calcification,and these pathways are directly or indirectly related to bone remodeling.This review provides a brief overview of the research progresses of vascular calcification-related molecular signaling pathways from the perspective of osteoblastic differentiation.

Effects of a Single Bout of Resistance Exercise in Different Volumes on Endothelium Adaptations in Healthy Animals / Efeitos de uma Sessão de Exercício Resistido em Diferentes Volumes sobre Adaptações Endoteliais em Animais Saudáveis

Abstract Background: Resistance exercise (RE) has been recommended for patients with cardiovascular diseases. Recently, a few studies have demonstrated that the intensity of a single bout of RE has an effect on endothelial adaptations to exercise. However, there is no data about the effects of different volumes of RE on endothelium function. Objective: The aim of the study was to evaluate the effects of different volumes of RE in a single bout on endothelium-dependent vasodilatation and nitric oxide (NO) synthesis in the mesenteric artery of healthy animals. Methods: Male Wistar rats were divided into three groups: Control (Ct); low-volume RE (LV, 5 sets x 10 repetitions) and high-volume RE (HV, 15 sets x 10 repetitions). The established intensity was 70% of the maximal repetition test. After the exercise protocol, rings of mesenteric artery were used for assessment of vascular reactivity, and other mesenteric arteries were prepared for detection of measure NO production by DAF-FM fluorescence. Insulin responsiveness on NO synthesis was evaluated by stimulating the vascular rings with insulin (10 nM). Results: The maximal relaxation response to insulin increased in the HV group only as compared with the Ct group. Moreover, the inhibition of nitric oxide synthesis (L-NAME) completely abolished the insulin-induced vasorelaxation in exercised rats. NO production showed a volume-dependent increase in the endothelial and smooth muscle layer. In endothelial layer, only Ct and LV groups showed a significant increase in NO synthesis when compared to their respective group under basal condition. On the other hand, in smooth muscle layer, NO fluorescence increased in all groups when compared to their respective group under basal condition. Conclusions: Our results suggest that a single bout of RE promotes vascular endothelium changes in a volume-dependent manner. The 15 sets x 10 repetitions exercise plan induced the greatest levels of NO synthesis.

Resumo Fundamentos: O exercício resistido (ER) tem sido recomendado para pacientes com doenças cardiovasculares. Recentemente, alguns estudos demonstraram que a intensidade de uma sessão de ER exerce um efeito sobre a disfunção endotelial. No entanto, não há dados sobre os efeitos de diferentes volumes de ER sobre a função endotelial. Objetivo: O objetivo deste estudo foi avaliar os efeitos de diferentes volumes de ER, realizados em uma única sessão, sobre a vasodilatação dependente do endotélio e síntese de óxido nítrico (NO) em artéria mesentérica de animais saudáveis. Métodos: Ratos Wistar machos foram divididos em três grupos: Controle (Ct); baixo volume (BV, 5 séries x 10 repetições) e alto volume de ER (AV, 15 séries x 10 repetições). Foi estabelecida a intensidade de 70% do teste de repetição máxima. Após o protocolo de exercício, anéis de artéria mesentérica foram utilizados na avaliação da reatividade vascular, e outras artérias mesentéricas foram preparadas para a detecção da produção de NO por fluorescência com para do DAF-FM. A resposta à insulina pela síntese de NO foi avaliada estimulando-se os anéis vasculares com insulina (10nM). Resultados: A resposta máxima do relaxamento induzido por insulina foi aumentada somente no grupo AV em comparação ao grupo Ct. Além disso, a inibição da síntese do NO (L-NAME), aboliu completamente o relaxamento vascular induzido por insulina em ratos exercitados. A produção de NO mostrou um aumento dependente do volume no endotélio e no músculo liso. No endotélio, apenas os grupos Ct e BV mostraram aumento significativo na síntese de NO quando comparado aos seus respectivos grupos sob condição basal. No entanto, no músculo liso, a fluorescência foi aumentada em todos os grupos quando comparados aos seus respectivos grupos sob a condição basal. Conclusões: Nossos resultados sugerem que uma única sessão de ER foi capaz de promover adaptações no endotélio vascular. Além disso, nós observamos que este efeito é volume-dependente e o volume de 15 séries x10 repetições induziu o maior aumento na síntese de NO.

Expression of TRPC1-BK Signal Complex in Aortic Smooth Muscle Tissue in Experimental Mice of Atherosclerosis / 中国循环杂志

Objective: To observe the expression of transient receptor potential channel-1—large conductance Ca2+-activated K+channel (TRPC1-BK) signal complex in aortic smooth muscle tissue in experimental mice of atherosclerosis (AS). Methods: Our research included in 2 groups: Control group, wild C57BL/6J mice were fed by normal diet and AS group, ApoE-/- mice were fed by high fat diet to establish AS model. All animals were treated for 10 weeks, n=10 in each group. The total RNA and protein were extracted from aortic smooth muscle tissue in sacrificed mice. The mRNA and protein expressions of TRPC1, BKα and BKβ1 subunit were measured by RT-PCR, Western blot analysis and immunohistochemistry staining. Results: By RT-PCR examination, compared with Control group, AS group showed increased mRNA expression of TRPC1, P<0.05 and decreased mRNA expressions of BKα and BKβ1 subunit, both P<0.01. By Western blot analysis, compared with Control group, AS group had elevated protein expression of TRPC1, P<0.05 and reduced protein expressions of BKα and BKβ1 subunit, both P<0.01; by immunohistochemistry staining, AS group had the higher protein expression of TRPC1, P<0.01 and the lower protein expressions of BKα and BKβ1 subunit, both P<0.05. Conclusion: The mRNA and protein expressions of TRPC1-BK signal complex were affected in aortic smooth muscle tissue in AS mice, which speculated that TRPC1-BK signal complex might be become a new target for treating the relevant vascular disease.

Bier's spots with onset in childhood

Abstract Bier spots are small, irregular, hypopigmented macules that are usually found on the arms and legs. The macules disappear when the limb is raised. Bier spots have been reported in association with a number of conditions but there is no consistent association to specific desease. Although they usually affect young adults, we report a case of Bier spots that began in childhood. As an asymptomatic and possibly transitional condition, the disease does not require treatment.

Jabuticaba-Induced Endothelium-Independent Vasodilating Effect on Isolated Arteries / Efeito Vasodilatador Independente do Endotélio Induzido pela Jabuticaba (M. cauliflora) em Artérias Isoladas

Abstract Background: Despite the important biological effects of jabuticaba, its actions on the cardiovascular system have not been clarified. Objectives: To determine the effects of jabuticaba hydroalcoholic extract (JHE) on vascular smooth muscle (VSM) of isolated arteries. Methods: Endothelium-denuded aortic rings of rats were mounted in isolated organ bath to record isometric tension. The relaxant effect of JHE and the influence of K+ channels and Ca2+ intra- and extracellular sources on JHE-stimulated response were assessed. Results: Arteries pre-contracted with phenylephrine showed concentration-dependent relaxation (0.380 to 1.92 mg/mL). Treatment with K+ channel blockers (tetraethyl-ammonium, glibenclamide, 4-aminopyridine) hindered relaxation due to JHE. In addition, phenylephrine-stimulated contraction was hindered by previous treatment with JHE. Inhibition of sarcoplasmic reticulum Ca2+ ATPase did not change relaxation due to JHE. In addition, JHE inhibited the contraction caused by Ca2+ influx stimulated by phenylephrine and KCl (75 mM). Conclusion: JHE induces endothelium-independent vasodilation. Activation of K+ channels and inhibition of Ca2+ influx through the membrane are involved in the JHE relaxant effect.

Resumo Fundamentos: Embora a jabuticaba apresente importantes efeitos biológicos, suas ações sobre o sistema cardiovascular ainda não foram esclarecidas. Objetivos: Determinar os efeitos do extrato de jabuticaba (EHJ) sobre o músculo liso vascular (MLV) em artérias isoladas. Métodos: Aortas (sem endotélio) de ratos foram montadas em banho de órgãos isolados para registro de tensão isométrica. Foram verificados o efeito relaxante, a influência dos canais de K+ e das fontes de Ca2+ intra- e extracelular sob a resposta estimulada pelo EHJ. Resultados: Artérias pré-contraídas com fenilefrina apresentaram relaxamento concentração-dependente (0,380 a 1,92 mg/mL). O tratamento com bloqueadores de canais de K+ (tetraetilamônio, glibenclamida, 4-aminopiridina) prejudicaram o relaxamento pelo EHJ. A contração estimulada com fenilefrina também foi prejudicada pelo tratamento prévio com EHJ. A inibição da Ca2+ATPase do reticulo sarcoplasmático não alterou o relaxamento pelo EHJ. Além disso, o EHJ inibiu a contração causada pelo influxo de Ca2+ estimulado por fenilefrina e KCl (75 mM). Conclusão: O EHJ induz vasodilatação independente do endotélio. Ativação dos canais de K+ e inibição do influxo de Ca2+ através da membrana estão envolvidas no efeito relaxante do EHJ.

Effect of large conductance Ca2+-activated K+ channel current and cytosolic calcium concentrations in retinal artery smooth muscle cells on diabetic retinal artery tension / 中华实验眼科杂志

Background Diabetic retinopathy (DR) is a common microvascular complications of the retina,retinal vascular smooth muscle cells of large conductance calcium-activated potassium channels (BK) is a major factor in regulating vasomotor and hemodynamic.Currently,functional changes of BK channel in retinal artery smooth muscle cells (RASMCs) and its role in DR were rarely reported.Objective This study was to investigate the early vascular damage mechanisms in DR by detecting the changes of BK channels current,calcium concentration and open probability (NP0) of BK channel with different calcium concentration in RASMCs of normal and diabetic rats.Method Fifty SPF SD 8-12 weeks old rats were randomly divided into normal control group and diabetic model group.Forty diabetic rats was intraperitoneally injected with 60 mg/kg streptozotocin to form type 1 diabetic model,10 rats (the normal control group) were injected sodium citrate solution with the same manner.Fluorescent probe was applied to detect calcium concentration in rat RASMCs;RASMCs were isolated by using enzyme digestion,and BK-channel electric currents and calcium concentrations in the RASMCs were measured by whole-cell patch clamp technique and fluorescence assay,respectively.The NP0 of BK channel was measured by single patch clamp technique.Results Diabetic models were successfully established in 36 rats with the success rate 90％.When stimulation voltage is greater than 60 mV,the current density of BK channel in RASMCs of diabetic model group decreased;when stimulating voltage was 100 mV,the BK channel currents of RASMCs in the normal control group and diabetic model group were (100±23) PA/PF and (50 ± 7) PA/PF,the difference was statistically significant (t =19.80,P ＜ 0.05).After adding specific BK channel blocker African scorpion toxin 100 nmol,the BK channel current in the normal control group significantly reduced,and that in the diabetes model group was not significantly changed;the calcium ion concentrations in RASMCs were (123±11)nmol/L and (255± 10)nmol/L in the normal control group and diabetic model group,the difference was statistically significant (t =32.50,P＜0.05).When stimulation voltage was 60 mV,with increasing calcium ion concentration,the NP0 of BK channel increased (F =15.28,P＜0.05).Conclusions The electric current and NP0 of BK-channel are obviously reduced and the calcium concentration is evidently elevated in RASMCs of diabetic rats,suggesting that the abnormal of BK-channel is probably one of the important causes of retinal artery abnormal contraction in diabetic rats.

Dynamic alteration of microRNA in high phosphorus induced calcification of vascular smooth muscle cell / 北京大学学报(医学版)

Objective:To study the change of microRNA during the early stage of high phosphorus in-duced vascular smooth muscle cell (VSMC)calcification and its related mechanism.Methods:The in vitro calcification model was created through stimulating VSMC cell line A7r5 with high Pi (2.6 mmol /L)for 7 d.The calcification was validated through ocresolphthalein complexone colorimetry to detect the cellular calcium content,real-time PCR to measure the calcification-related gene expression and alizarin red staining to observe the formation of calcium nodules.Based on the cell calcification model,micro-RNA microarray array was applied to screen the profiles of microRNA expression in VSMC following high Pi stimulation for different periods (0,3 and 12 h).The array data were analyzed by TAMtool to explore the activated signaling pathway.Results:The calcium content of A7r5 cells induced by high Pi was in-creased 9.6 times high as cells without Pi treatment (P <0.05 ).VSMC contractile phenotype genes (SM-αactin,SM22)were down-regulated (P <0.05 ),while calcification-related genes (BMP2, MSX2,Runx2)were up-regulated (P <0.05)in VSMC stimulated by high Pi.The calcium nodules were obviously formed in cells after 7 d high Pi treatment.In microarray experiment,680 individual mi-croRNAs were detected in high Pi-treated VSMCs at different time points (0,3 and 12 h).Among these genes,miR-183,miR-664 and miR-9 * were increased whereas miR-542-5P,let-7f and miR-29a were decreased in time-dependent manners.Twenty-six kinds of signaling pathways,including cell apoptosis, differentiation and proliferation,were significantly activated.All these activated pathways were associated with calcification.Conclusion:This study implies that microRNA changed in high Pi-induced VSMCs may involve in the process of calcification.

Effects of One Resistance Exercise Session on Vascular Smooth Muscle of Hypertensive Rats / Efeitos de uma Sessão de Exercício Resistido sobre o Músculo Liso Vascular de Ratos Hipertensos

AbstractBackground:Hypertension is a public health problem and increases the incidence of cardiovascular diseases.Objective:To evaluate the effects of a resistance exercise session on the contractile and relaxing mechanisms of vascular smooth muscle in mesenteric arteries of NG-nitro L-arginine methyl ester (L-NAME)-induced hypertensive rats.Methods:Wistar rats were divided into three groups: control (C), hypertensive (H), and exercised hypertensive (EH). Hypertension was induced by administration of 20 mg/kg of L-NAME for 7 days prior to experimental protocols. The resistance exercise protocol consisted of 10 sets of 10 repetitions and intensity of 40% of one repetition maximum. The reactivity of vascular smooth muscle was evaluated by concentration‑response curves to phenylephrine (PHEN), potassium chloride (KCl) and sodium nitroprusside (SNP).Results:Rats treated with L-NAME showed an increase (p < 0.001) in systolic blood pressure (SBP), diastolic blood pressure (DBP) and mean arterial pressure (MAP) compared to the initial period of induction. No difference in PHEN sensitivity was observed between groups H and EH. Acute resistance exercise reduced (p < 0.001) the contractile response induced by KCl at concentrations of 40 and 60 mM in group EH. Greater (p < 0.01) smooth muscle sensitivity to NPS was observed in group EH as compared to group H.Conclusion:One resistance exercise session reduces the contractile response induced by KCl in addition to increasing the sensitivity of smooth muscle to NO in mesenteric arteries of hypertensive rats.

ResumoFundamento:A hipertensão é um problema de saúde pública e faz aumentar a incidência das doenças cardiovasculares.Objetivo:Avaliar os efeitos de uma sessão de exercício resistido sobre os mecanismos contráteis e relaxantes do músculo liso vascular em artéria mesentérica de ratos hipertensos induzidos por L-NAME.Métodos:Ratos Wistar foram divididos em três grupos: Controle (C), Hipertenso (H) e Hipertenso Exercitado (HE). A hipertensão foi induzida pela administração de 20 mg/kg de NG-nitro L-arginina metil éster (L-NAME) durante sete dias antes dos protocolos experimentais. O protocolo de exercício resistido consistiu em dez séries de dez repetições e intensidade de 40% de uma repetição máxima. A reatividade do músculo liso vascular foi avaliada através de curvas concentração-resposta para a fenilefrina (FEN), cloreto de potássio (KCl) e nitroprussiato de sódio (NPS).Resultados:Os ratos tratados com L-NAME apresentaram aumento (p < 0,001) da Pressão Arterial Sistólica (PAS), da Pressão Arterial Diastólica (PAD) e da Pressão Arterial Média (PAM) quando comparados ao período inicial da indução. Não foi observada diferença na sensibilidade da FEN entre os grupos H e HE. O exercício resistido agudo reduziu (p < 0,001) a resposta contrátil induzida pelo KCl nas concentrações de 40 e 60 mM do grupo HE quando comparado ao grupo H. Foi observado maior (p < 0,01) sensibilidade do músculo liso ao NPS no grupo HE quando comparado ao grupo H.Conclusão:Uma sessão de exercício resistido reduz as respostas contráteis induzidas pelo KCl, além de aumentar a sensibilidade do músculo liso ao NO em artéria mesentérica de ratos hipertensos.

Renal hemangiopericytoma: case report and literature review / Hemangiopericitoma renal: relato de caso e revisão de literatura

Hemangioperycytoma is a rare perivascular tumor that seldom involves the urogenital system. This tumor often appears with an unspecific clinical picture, and sometimes is associated with hematuria or hypertension. Diagnosis is based on a combination of histological and immunohistological findings. We report a case of a 52-year-old patient with renal hemangiopericytoma who underwent surgical treatment at our service. This report also includes a literature review on the subject.

Hemangiopericitoma é um raro tumor perivascular que raramente envolve o sistema urogenital. Esses tumores geralmente se manifestam com quadro clínico inespecífico, por vezes associado a hematúria ou hipertensão. O diagnóstico baseia-se numa combinação de alterações histológicas e imuno-histológica. Este artigo relatou o caso de uma paciente de 52 anos de idade com um hemangiopericitoma renal submetida a tratamento cirúrgico em nosso serviço e incluiu uma revisão de literatura sobre o assunto.

The effects of Slit2 protein on proliferation and migration of rats vascular smooth muscle cells stimulated by TNF-α / 重庆医学

Objective To observe the effects of Slit2 protein on the proliferation and migration of VSMCs .Methods The VSMCs was cultured in our laboratory .The experiment was divided into two parts ,part one:VSMCs were divided into normal con‐trol group and experimental groups(culture with 50 ,75 ,100 ,125 and 150 ng/mL Slit2 respectively);part two :VSMCs were divided into normal control group ,positive control group(culture with TNF‐α10 ng/mL) and experimental groups(culture with TNF‐α10 ng/mL+Slit2 50 ng/mL ,TNF‐α10 ng/mL+Slit2 75ng/mL ,TNF‐α10 ng/mL+ Slit2 100 ng/mL ,TNF‐α10 ng/mL+ Slit2 125 ng/mL and TNF‐α10 ng/mL+Slit2 150 ng/mL respectively) .To detect proliferation and migration of VSMCs by CCK‐8 and tran‐swell experiment .Results The difference of OD value and numbers of VSMCs has no statistical significance in the presence of Slit 2 (P=0 .516 ,P=0 .52) .The numbers of VSMCs has statistical significance between control and positive control groups (P=0 .00) . The numbers of VSMCs in experimental groups were fewer than positive control group (P<0 .05) ,whereas the difference of OD value still has no statistical significance between experimental and positive groups (P= 0 .173) .Conclusion Recombinant Slit2 could inhibits migration in VSMCs induced by TNF‐α,whereas it has no effect on proliferation of VSMCs .

Effect of aging on transcription and protein expressions of procollagen α polypeptide gene of vascular smooth muscle cells in rat / 中华老年医学杂志

Objective To investigate the effects of aging on procollagen α polypeptide gene transcription and protein expression in rat vascular smooth muscle cells.Methods Vascular smooth muscle cells from thoracoabdominal aorta in neonate and 9 months old healthy Wistar rats were cultured in vitro.Results Transcription polymerase chain reaction(RT PCR) and Western blotting were used to detect type Ⅰ and Ⅲ pro-collagen α polypeptide mRNA and protein.The RT-PCR displayed that type Ⅰ procollagen α polypeptide mRNA expression had no significant difference between young group and adult group [(76.62±1.05) vs.(78.37±2.42),P＞0.05].Type Ⅲ procollagen α polypeptide mRNA expression was (105.40 ± 2.66) in young group and (123.10 ± 3.81) in adult group(P＞0.05).Type Ⅰ procollagen α polypeptide mRNA expression was (3.13 ±0.54) in young group and (4.63 ± 1.03) in adult group (P=0.05).Type Ⅲ procollagen α polypeptide mRNA expression had no significant difference between the adult and young groups[(6.86 ±0.41) vs.(7.68±0.63),P＞0.05].Type Ⅰ and type Ⅲ procollagen α polypeptide protein expressions were increased significantly in adult group as compared with the young group [(0.10 ± 0.03) vs.(0.06±0.03),(0.58±0.06) vs.(0.40±0.02),both P＜0.05].Conclusions Aging increases the procollagen α polypeptide level in vascular smooth muscle cell,which may involve in the development of vascular remodeling and atherosclerosis.

Study on the action mechanism of basic fibroblast growth factor on vascular smooth muscle cells in spontaneously hypertensive rats / 中华老年医学杂志

Objective To explore the action mechanism of basic fibroblast growth factor (bFGF) on vascular smooth muscle cells (VSMCs) in spontaneously hypertensive rats (SHR).Methods Ts were obtained from SHR and SD rats.The aortic VSMCs were cultured in vitro by tissue explant method.VSMCs were treated with different concentration of exogenous bFGF (0 ng/ml,20 ng/ml,40 ng/ml,60 ng/ml,80 ng/ml,100 ng/ml) for 48 h,then cell proliferation was detected by 3 (4,5 dimethylthiazol)2,5 diphenyltetrazolium bromide (MTT) colorimetric assay.VSMCs from SHR in control group were treated with bFGF (100 ng/ml) for 48h.VSMCs from SHR in treatment group were treated with bFGF (100 ng/ml) plus Proteinkinase C(PKC) inhibitor (staurosporine) for 48 h.Results After treatment with different concentration (0 ng/ml,20 ng/ml,40 ng/ml,60 ng/ml,80 ng/ml,100 ng/ml) of bFGF for 48 h,the values measured by MTT colorimetric method were 0.402 ± 0.103,0.605 ±0.090,0.696 ± 0.131,0.812 ± 0.080,0.901 ± 0.065,1.056±0.078 respectively in aortic VSMCs from SD rats,and 0.404±0.065,0.507±0.078,0.608±0.057,0.704 ± 0.107,0.812 ± 0.097,0.908 ± 0.032 respectively in aortic VSMCs from SHR.Compared with control group,the values measured by MTT colorimetric method were decreased in treatment group (P＜0.05).The proliferative effect of bFGF in aortic VSMCs from SHR was attenuated after administration of PKC inhibitor staurosporine.Conclusions Exogenous bFGF administration promotes VSMCs proliferation in SHR and SD rats in a concentration-dependent manner.PKC plays an important role in the signal transduction mechanism in VSMCs proliferation by exogenous bFGF.