联合培养的成纤维细胞对平滑肌细胞增殖和形态的影响

目的:观察体外联合培养的成纤维细胞对平滑肌细胞形态和增殖的影响,为研究成纤维细胞和平滑肌细胞生物学特性和相互关系以及体外构建含此两种细胞的组织工程皮肤提供理论和实践基础。方法:在体外分别分离培养成纤维细胞和平滑肌细胞并常规传代培养。模拟组织工程皮肤的结构及成纤维细胞和平滑肌细胞间的相互影响途径,按照不同比例(平滑肌细胞:成纤维细胞=1:1,1:2,1:3)建立以胶原凝胶为基质的成纤维细胞与平滑肌细胞联合培养模型。应用荧光标记、组织学观察和MTT法对平滑肌细胞的形态和代谢进行研究。结果:联合培养模型法可以有效地建立起成纤维细胞和平滑肌细胞的相互影响模式。1:1组的成纤维细胞对联合培养的平滑肌细胞增殖有促进作用,1:3组的促增殖作用则不明显。结论:1:1联合培养的成纤维细胞对平滑肌细胞增殖有促进作用,对它们体外培养和扩增以及相互关系的研究对阐明这两种细胞组合作为真皮替代物种子细胞具有重要的意义。     

Aortic endothelial and smooth muscle cell co-culture: an in vitro model of the arterial wall.

Interactions between vascular endothelial (EC) and smooth muscle cells (SMC) contribute both to the normal function of the vascular wall and to the pathogenesis of lesions such as atherosclerosis and fibrointimal hyperplasia. However, study of these interactions has been hampered by the difficulty in growing these two cell types in simultaneous culture. Methods using conditioned media, shared media, and bilayer culture have been described, but none is well suited to the study of vascular cell interactions. We report a method for EC-SMC co-culture that preserves bilayer morphology, allows independent study of the cells and their matrices after intervention, remains stable over long periods in culture, and permits study of changes in cell-cell interaction with growth of the cells to confluence. This simple bilayer co-culture system simulates the in vivo situation and may enhance our understanding of EC-SMC interactions.     

体外共培养体系中胆管癌细胞对内皮细胞的作用

目的 在体外共培养体系中研究胆管癌细胞对内皮细胞的作用.方法 建立胆管癌QBC939细胞与内皮细胞的体外共培养体系设为共培养组,同期单独培养的内皮细胞设为内皮细胞组,同样数量的内皮细胞和胆管癌细胞分别单独培养的上清液混合液设为混合组.对内皮细胞进行光镜及扫描电镜观察,并用免疫荧光法检测内皮细胞蛋白水解酶pp125焦点激酶(pp125FAK)、基质金属蛋白酶2和9(MMP-2、MMP-9)、人尿激酶型纤维蛋白溶解酶原激活物(uPA)表达的变化,明胶酶谱法测定内皮细胞MMP-2及MMP-9的活性.采用配对t检验分析其结果.结果 与内皮细胞组比较,共培养组内皮细胞之间的间隙增大.内皮细胞组ppl25FAK、MMP-2、MMP-9、uPA的平均荧光强度分别为394±51、455±82、377±48、422±55,而共培养组表达增强,分别为1096 ±128、931 ±72、815 ±76、801 ±56,两组比较差异有统计学意义(t=6.53,4.32,3.61,3.45,P<0.05).混合组MMP-2、MMP-9灰度扫描值分别为240.2±15.2和2.4±0.8.共培养组的MMP-2、MMP-9分别为687.4±43.6和150.9±13.2,两组比较差异有统计学意义(t=4.89,5.43,P<0.05).结论 共培养后的内皮细胞之间缝隙增大,蛋白水解酶表达增强,它可能参与降解内皮细胞外基质,促进了胆管癌的转移.     

Differential effects of orbital and laminar shear stress on endothelial cells

OBJECTIVE: Laminar shear stress is atheroprotective for endothelial cells (ECs), whereas nonlaminar, disturbed, or oscillatory shear stress correlates with development of atherosclerosis and neointimal hyperplasia. The effects of orbital and laminar shear stress on EC morphology, proliferation, and apoptosis were compared. METHODS: ECs were exposed to orbital shear stress with an orbital shaker (210 rpm) or laminar shear stress (14 dyne/cm 2) with a parallel plate. Shear stress in the orbital shaker was measured with optical velocimetry. Cell proliferation was assessed with direct counting and proliferating cell nuclear antigen staining; apoptosis was assessed with transferase-mediated deoxyuridine triphosphate nick end labeling staining. Cell surface E-selectin and intercellular adhesion molecule expression were assessed with fluorescence-activated cell sorting. Akt phosphorylation was assessed with Western blotting. RESULTS: Orbital shear stress increased EC proliferation by 29% and 3 [H]thymidine incorporation two-fold compared to 16% and 38% decreases, respectively, in ECs treated with laminar shear stress (P < .0001 and P = .03, analysis of variance). Cells in the periphery of the culture well aligned to the direction of shear stress similar to the shape change seen with laminar shear stress, whereas ECs in the center of the well appeared unaligned similar to ECs not exposed to shear stress. Shear stress at the bottom surface of the culture well was reduced in the center of the well (5 dyne/cm 2) compared to the periphery (11 dyne/cm 2); the Reynolds' number was 2066. ECs were seeded differentially in the center and periphery of the wells. ECs in the center of the well had increased proliferation, increased apoptosis, reduced Akt phosphorylation, increased intercelluar adhesion molecule expression, and reduced E-selectin down-regulation, compared with ECs in the periphery of the well. CONCLUSION: Although the orbital shaker does not apply uniform shear stress throughout the culture well, arterial magnitudes of shear stress are present in the periphery of the well. ECs cultured in the center of the well exposed to low magnitudes of orbital shear stress might be a model of the "activated" EC phenotype. CLINICAL RELEVANCE: The perfect in vitro model to study and assess treatments for atherosclerosis and neointimal hyperplasia does not exists. An extensive body of literature describing effects of laminar shear stress on endothelial cells has contributed to our understanding of the interactions between shear stress and blood vessels. Laminar shear stress is atheroprotective, whereas oscillatory or disturbed shear stress correlates with areas of atherosclerosis and neointimal hyperplasia in vivo. This study describes the orbital shear stress model, its effects on endothelial cell proliferation and apoptosis, and suggests that activation of the intracellular Akt pathway is associated with these differing effects of laminar and orbital shear stress on endothelial cells.     

Vascular endothelial growth factor increases the migration and proliferation of smooth muscle cells through the mediation of growth factors released by endothelial cells

BACKGROUND: Vascular endothelial growth factor (VEGF), a highly specific chemotactic and mitogenic factor for vascular endothelial cells (EC), appears to be involved in the development of atherosclerosis. The purpose of our study was to assess if VEGF might indirectly stimulate SMC migration and proliferation in a EC-SMC coculture system, through the mediation of growth factors released by EC. METHODS: Bovine aortic SMC were cocultured with bovine aortic EC treated with hrVEGF, to assess SMC proliferation and migration. The release and mRNA expression of basic fibroblast growth factor (bFGF) and transforming growth factor beta(1) (TGFbeta(1)) were assessed by ELISA and PCR analysis. RESULTS: hrVEGF (10 ng/ml), added to EC cocultured with SMC, induced a significant increase in tritiated thymidine uptake by SMC as compared to controls (P < 0.01) and a significant increase in SMC migration in respect to control (27%; P < 0.01). EC stimulated with hrVEGF increased the release and the expression of bFGF and decreased the release and the expression of TGFbeta(1) with a statistically significant difference in respect to controls (P < 0.001). CONCLUSIONS: VEGF indirectly stimulates SMC proliferation and migration through the modulation of bFGF and TGFbeta(1) released by EC.     

鼠双微体-2基因在醛固酮增多症大鼠模型主动脉平滑肌中的表达

目的 观察醛固酮对大鼠主动脉平滑肌鼠双微体-2基因(MDM2)表达的影响.方法 将SD大鼠随机分为3组,每组8只.采用皮下给药的方法建立醛固酮增多症模型,其中醛固酮组经微量渗透泵持续释放醛崮酮(1μg/h);醛同酮+螺内酯组除给予等量醛固酮外,每日行螺内酯灌胃(每日100mg/kg);对照组仅泵空白溶剂.尾套法检测大鼠血压,4周后处死所有大鼠,测定血钾、钠、醛固酮浓度及血浆肾素活性.分别采用逆转录.聚合酶链反应(RT-PCR)、Westerill blot、免疫组织化学检测主动脉平滑肌MDM2基因表达.结果 (1)成功建立醛同酮增多症大鼠模型:醛固酮组2周后血压明显升高,血钾下降,呈现低肾素、高醛固酮特征,与对照组比较差异有统计学意义(P<0.01);(2)醛固酮组主动脉平滑肌MDM2基因表达明显高于对照组(P<0.01).螺内酯可抑制醛固酮的上述作用(P<0.01).结论 醛固酮可促进血管平滑肌细胞中MDM2的表达,而螺内酯可拮抗其效应.     

apelin-13穿越非对称性二甲基精氨酸损伤的血管内皮层对血管平滑肌细胞的作用

目的 探讨尿毒症毒素非对称性二甲基精氨酸（ADMA）致内皮完整性破坏的情况下,血管活性肽apelin-13对血管平滑肌细胞收缩作用的影响。方法 利用Transwell小室建立单层内皮细胞屏障结构,分别设立实验组和对照组。实验组的内皮细胞经ADMA刺激后,在两组的上室中加入FITC标记的apelin-13,测出不同时间下室apelin-13的浓度,计算通透系数Pa值,并通过免疫荧光染色观察内皮细胞形态的变化。随后,利用Transwell小室建立上室内皮细胞、下室平滑肌细胞的双室模型。设立4个实验组：空白对照组、单纯ADMA作用组（ADMA组）、apelin-13穿越组（apelin组）、ADMA作用后apelin-13穿越组（ADMA+ apelin组）。内皮细胞经ADMA刺激后,于上室中加入apelin-13,分别用免疫荧光及Western印迹法对平滑肌细胞中磷酸化肌球蛋白轻链（p-MLC）进行定性及定量检测。 结果 ADMA可改变内皮细胞骨架及细胞间连接的结构,并显著增加单层内皮细胞对apelin-13的通透性。实验组与对照组的Pa百分比随apelin-13穿越时间而改变,在20 min时达最大值,与0时间点差异有统计学意义[（176.3±9.2）%比（100.3±1.5）%,P < 0.05]。在双室模型中,ADMA+apelin组p-MLC表达量最高,与apelin组差异有统计学意义,ADMA组中也有少量p-MLC表达,3组与空白对照组差异均有统计学意义。 结论 ADMA可通过影响细胞骨架及细胞间连接,导致单层血管内皮细胞对apelin-13的通透性增高。apelin-13穿越损伤的内皮层引起平滑肌p-MLC水平升高,可能参与了尿毒症高血压的发生。     

Induction of endostatin expression in meniscal fibrochondrocytes by co-culture with endothelial cells

Introduction  The ultimate goal after meniscus damage is the preservation of the original meniscal tissue, which is often impossible due to the limited healing capacity of meniscal lesions, especially in the avascular zone. Factors produced by endothelial cells of meniscal vessels may contribute to better wound healing in vascularized zones. We therefore investigated the expression of different angiogenic factors, growth hormones and cytokines in human fibrochondrocytes and in fibrochondrocytes upon co-culture with endothelial cells, to examine mechanisms of repair of meniscal injury in more detail and to investigate the potential use of endothelial cells in co-cultures for autologous meniscal repair utilizing tissue engineering technology. Materials and methods  Gene expression of SMAD-4, iNOS, IL-1β, VEGF, MMP-1, MMP-3, MMP-13, aggrecan, biglycan, vimentin, collagen-I, -II, -III, -IV, -VI, -X, -XVIII, angiopoietin-1, angiopoietin-2, and thrombostatin-1 were investigated in fibrochondrocytes in comparison to cells in co-culture with human umbilical vein endothelial cells (HUVEC). The expression of endostatin was enumerated in cell supernatants. A proliferation assay was used to investigate the mitotic activity of the cells. Results  In presence of HUVEC, meniscal fibrochondrocytes expressed SMAD-4, iNOS, IL-1β, VEGF, MMP-1, MMP-3, MMP-13, aggrecan, biglycan, vimentin, collagen-I, -II, -III, -VI, and -XVIII at rates comparable to cells without HUVECs. Note that the expression of endostatin was significantly higher in the co-culture when compared to the separate fibrochondrocyte cultures and the proliferation rate of endothelial cells was significantly decreased in co-culture. Conclusion  We conclude that the expression of the anti-angiogenic factor endostatin increased in the fibrochondrocytes. This may limit the regeneration capacities of meniscal injury in vivo.     

反义寡核苷酸对血管平滑肌细胞c-myc、增殖细胞核抗原蛋白表达的影响

目的　研究反义c myc、增殖细胞核抗原 (PCNA)基因片断对血管平滑肌细胞 (VSMCs)增殖的作用。　方法　使正义、反义及错配反义c myc、PCNA ,分别作用于体外培养的VSMCs ,应用蛋白印迹和免疫组化法测定并经计算机图像分析 ,观察其对c myc、PCNA蛋白表达的影响。　结果　 10μmol/L浓度的反义c myc、PCNA作用于VSMCs后 1～ 5d ,相应c myc和PCNA蛋白表达减弱 ,计算机图像分析表明其表达产物与对照组相比 ,差异有显著意义 (P <0 0 5 ) ,而正义和错配反义寡核苷酸无此效应 (P >0 0 5 )。　结论　反义c myc、PCNA寡核苷酸可通过阻遏其相应基因表达进而抑制蛋白表达及VSMCs增殖。     

The effects of thrombin on bovine aortic endothelial and smooth muscle cells

Recent evidence suggests that thrombin interacts with various cell types, stimulating cellular proliferation and protein and prostanoid production. To further delineate its role in vascular healing, we have studied the effects of thrombin on proliferation and matrix production by the cells of the vessel wall. The addition of thrombin (1 unit/ml) to cultures of bovine aortic smooth muscle cells resulted in an increase in cell proliferation (p less than 0.01) and number (p less than 0.03), whereas in cultures of bovine aortic endothelial cells thrombin produced a decrease in cell proliferation (p less than 0.01) and number (p less than 0.02). Thrombin also altered matrix composition in cultures of these cells. In both bovine aortic endothelial cells and bovine aortic smooth muscle cell cultures grown in the presence of thrombin, total protein content was significantly increased when compared to controls (p less than 0.03). In bovine aortic endothelial cell cultures the addition of thrombin resulted in a decrease in collagen content (p less than 0.01) and an increase in sulfated glycosaminoglycan content (p less than 0.02). In contrast, in bovine aortic smooth muscle cell cultures thrombin resulted in an increase in collagen content (p less than 0.03), whereas glycosaminoglycan content was unaffected. These findings suggest that thrombin may significantly influence vascular healing and function by altering cell number and matrix composition.     

高磷对血管平滑肌细胞骨钙素mRNA表达和钙沉积的影响

目的 观察高磷对培养的牛主动脉平滑肌细胞骨钙素(OC)mRNA表达和钙沉积的影响,探讨高磷是否为促进血管钙化的独立因素。方法 用不同磷浓度[正常组(Pi 1.5 mmol/L)、高磷组(2.0 mmol/L)]的培养液,培养牛主动脉平滑肌细胞,观察72 h骨钙素的表达,以及不同时间(第3、6、9天)血管平滑肌细胞的钙沉积。上清液中骨钙素浓度用放射免疫法测定。RT-PCR观察骨钙素mRNA的表达。平滑肌细胞钙含量用甲O-酚酞络合酮方法测定。BCA法测定蛋白含量,用蛋白含量标化骨钙素浓度和钙含量。结果 3d后与正常磷组相比,高磷组上清液骨钙素水平明显增高[高磷组(15.03±2.60)pg/μg蛋白比正常磷组(2.98±0.84)pg/μg蛋白,P<0.001];高磷组平滑肌细胞骨钙素mRNA的表达也显著增加(OC/GAPDH:1.91±0.13比0.75±0.04,P<0.001)。高磷组钙沉积显著增加[培养第6天,高磷组(77.19±11.69)μg/mg蛋白比正常磷组(25.77±1.75)μg/mg蛋白,P<0.01],呈时间和剂量依赖性。von Kossa钙染色,高磷组平滑肌细胞中见大量的黑色颗粒沉积。结论 高磷可直接刺激平滑肌细胞的钙沉积和骨钙素产生增加,促进血管平滑肌细胞钙化。高磷血症是促发血管钙化的独立因素。     

Diabetes attenuates the alterations in both venous endothelial and smooth muscle cell function induced by prolonged hypercholesterolemia in an experimental model.

The systemic effects of the combination of diabetes and hypercholesterolemia on venous vasomotor function are poorly understood. This study examines in vitro vasomotor responses of New Zealand White rabbit jugular veins from control, diabetic, hypercholesterolemic, and hypercholesterolemic with diabetes groups. Hypercholesterolemia was induced with a diet supplemented with 1% cholesterol, while diabetes was induced by alloxan. Cumulative dose response curves to norepinephrine, bradykinin, and histamine were performed. After precontraction with norepinephrine to give 80% maximal contraction, relaxation in response to acetylcholine and sodium nitroprusside was determined. Potency of the agonist responses were compared. The contractile responses to all agonists were significantly increased in hypercholesterolemia. Only the response to norepinephrine was increased in diabetes. However, when diabetes and hypercholesterolemia were combined the contractile response to bradykinin was increased, the response to histamine was significantly decreased, but the norepinephrine response was unchanged. There was dose-dependent, endothelium-mediated relaxation in precontracted control and diabetic veins. Hypercholesterolemia and hypercholesterolemia with diabetes interfered with endothelium-mediated relaxation, producing a multiphasic response: relaxation at low concentrations followed by contraction at higher concentrations. Non-endothelium-dependent relaxation to sodium nitroprusside of precontracted veins was unaffected by the presence of diabetes or hypercholesterolemia. This study suggests that the combined presence of diabetes and hypercholesterolemia attenuated the altered contractile responses induced by hypercholesterolemia alone without further alterations in endothelial-mediated responses. The mechanism for these alterations remains to be determined.     

Differential responsiveness of early- and late-passage endothelial cells to shear stress

BACKGROUND: The incidence of vascular disease increases with age. Because atherosclerosis and neointimal hyperplasia colocalize in areas of disturbed shear stress, the effects of orbital shear stress (SS) on endothelial cell proliferation, protein kinase B (Akt) activation, and functional activity were analyzed using a senescence model. METHODS: Early- (p3 to 7) and late- (p28 to 32) passage bovine aortic endothelial cells were exposed to orbital SS (210 rpm) or static conditions (0 to 5 days). Cell proliferation was directly counted and confirmed with proliferating cell nuclear antigen reactivity. Phosphorylated and total Akt were assessed with Western blotting. Endothelial cell-induced smooth muscle cell migration was assessed with a Boyden chamber. RESULTS: Late-passage endothelial cells demonstrated no increase in orbital SS stimulated proliferation compared with early-passage cells (P = .42). Late-passage endothelial cells demonstrated decreased Akt phosphorylation in response to SS compared with early passage cells (n = 6, P = .01). Late-passage cells induced 26% less smooth muscle cell migration than early-passage cells (n = 3, P = .03). CONCLUSIONS: Late-passage endothelial cells demonstrate decreased proliferation, Akt phosphorylation, and secretion of smooth muscle cell chemoattractants in response to orbital SS compared with early passage cells. These results suggest that late-passage endothelial cells respond to SS differently than early-passage cells and confirm the utility of the in vitro senescence model.     

轻度血尿酸升高对大鼠肾小球内皮细胞及血管平滑肌细胞功能的影响

目的 通过观察轻度血尿酸升高对大鼠肾小球内皮细胞功能损伤及血管平滑肌细胞增殖的影响,探讨轻度血尿酸升高是否能导致肾脏损害及降尿酸治疗对肾脏的保护作用.方法 用雄性SD大鼠为研究对象,随机分为4组:对照组(n=15)、氧嗪酸组(n=15)、别嘌呤醇组(n=12)和氧嗪酸+别嘌呤醇组(n=12).予以低盐饮食,每隔10天监测各组大鼠的动脉血压.于试验后20 d及40 d用ELISA法分别测定各组大鼠内皮细胞功能受损的指标[一氧化氮(NO)、内皮素(ET)1、纤溶酶原激活物(PAI)1]、血管平滑肌细胞增殖的指标[血小板衍生因子(PDGF)、环氧化酶(COX)2、单核细胞趋化蛋白(MCP)1的含量]以及炎性反应指标[白细胞介素(IL)18、肿瘤坏死因子(TNF)α];同时观察各组大鼠肾功能及肾脏组织病理变化;免疫组化法检测各组大鼠肾组织中PDGF、一氧化氮合酶(NOS)的表达.结果 与对照组相比,氧嗪酸组大鼠血浆NO浓度显著降低(P＜0.05),ET-1、PAI-1、PDGF、MCP-1、COX2、TNF-α、IL-18浓度均显著升高(均P＜ 0.05).光镜下,各组大鼠肾组织均未见尿酸结晶形成,氧嗪酸组肾小血管管壁增厚,内膜增生,管腔狭窄;免疫组化结果显示,与对照组相比,氧嗪酸组NOS的表达显著减少(7.33％±2.11％比25.75％±2.33％,P＜0.05),PDGF的表达显著增多(31.18％±2.83％比8.09％±1.81％,P＜ 0.05).经别嘌呤醇降尿酸干预治疗后大鼠血清中内皮细胞损伤指标NO上调(P＜0.05),而ET-1及PAI-1均下调(均P＜0.05);而血管平滑肌增殖指标及炎性指标均下调(均P＜ 0.05).结论 轻度血尿酸升高可导致肾小球内皮细胞功能受损、血管平滑肌细胞增殖;降尿酸治疗能改善内皮细胞功能,减轻血管平滑肌细胞的增殖.     

Transfer and expression of the interferon gamma gene in human endothelial cells inhibits vascular smooth muscle cell growth in vitro

Intimal hyperplasia in blood vessels is primarily caused by the migration and proliferation of vascular smooth muscle cells. Excessive intimal thickening characterizes atherosclerosis as well as bypass graft and angioplasty failures. Endothelial cell-smooth muscle cell interactions and local cytokine production are important regulators of smooth muscle cell growth. Interferon gamma (γ-IFN), a product of T lymphocytes found in atherosclerotic lesions, inhibits smooth muscle cell proliferation in vitro. To determine if local delivery of γ-IFN may be useful in the treatment or prevention of vascular proliferative diseases, we transferred the human γ-IFN gene into endothelial cells isolated from human arteries and microvessels using a retroviral vector. Biologically active γ-IFN was produced and secreted by γ-IFN transduced endothelial cells, but not by control, nontransduced cells, or cells identically transduced with E. coli beta galactosidase (β-gal). To more closely approximate the microenvironment of blood vessels, subconfluent smooth muscle cells were plated in coculture with control, nontransduced endothelial cells, γ-IFN transduced endothelial cells, or β-gal transduced endothelial cells. Smooth muscle cell growth was inhibited 30–70% by coculture with γ-IFN transduced endothelial cells compared to coculture with β-gal transduced or control endothelial cells (p < 0.05). Our results suggest endothelial cells modified to produce γ-IFN may be a useful therapy in proliferative vascular diseases.     

剪切力对支架边缘内皮细胞的生物学影响

经皮冠状动脉介入(PCI)是治疗冠心病重要手段,但其不可避免的会损伤内皮细胞,引起内皮细胞功能失调和/或结构损害。血管内皮细胞在冠状动脉支架再狭窄中扮演了非常重要角色,而一直作用于内皮的血液流动剪切力对内皮细胞形态及功能的改变起重要作用,本文就剪切力对支架边缘内皮细胞的生物学影响研究概况作一综述。     

Lidocaine attenuates cytokine-induced cell injury in endothelial and vascular smooth muscle cells

Local anesthetics have been reported to attenuate the inflammatory response and ischemia/reperfusion injury. Therefore, we hypothesized that pretreatment with local anesthetics may protect endothelial and vascular smooth muscle (VSM) cells from cytokine-induced injury. Human microvascular endothelial cells and rat VSM cells were pretreated with lidocaine or tetracaine (5-100 microM for 30 min) and then exposed to the cytokines tumor necrosis factor-alpha, interferon-gamma, and interleukin-1beta for 72 h. Cell survival and integrity were evaluated by trypan blue exclusion and lactate dehydrogenase release. The role of adenosine triphosphate-sensitive potassium (KATP) channels, protein kinase C, or both in modulating local anesthetic-induced protection was evaluated with the mitochondrial KATP antagonist 5-hydroxydecanoate, the cell-surface KATP antagonist 1-[5-[2-(5-chloro-o-anisamido)ethyl]-2-methoxyphenyl]sulfonyl-3-methylthiourea (HMR-1098), and the protein kinase C inhibitor staurosporine. Lidocaine attenuated cytokine-induced cell injury in a dose-dependent manner. Lidocaine (5 microM) increased cell survival by approximately 10%, whereas lidocaine (100 microM) increased cell survival by approximately 60% and induced a threefold decrease in lactate dehydrogenase release in both cell types. In contrast, tetracaine did not attenuate cytokine-induced cell injury. 5-hydroxydecanoate abolished the protective effects of lidocaine, but staurosporine and HMR-1098 had no effect on the lidocaine-induced protection. This study showed that lidocaine, but not tetracaine, attenuates cytokine-induced injury in endothelial and VSM cells. Lidocaine-induced protection appears to be modulated by mitochondrial KATP channels. IMPLICATIONS: This study demonstrates that lidocaine attenuates cytokine-induced injury of endothelial and vascular smooth muscle cells via mechanisms involving adenosine triphosphate-sensitive potassium channels. Protection of the vasculature from cytokine-induced inflammation may preserve important physiological endothelial and vascular smooth muscle functions.