miR-145抑制VSMC增殖的作用及其相关信号通路研究

目的探讨miR-145对PDGF-BB诱导的大鼠原代血管平滑肌细胞(VSMC)的作用及丝裂原激活的蛋白激酶(MAPK)信号转导途径的作用。方法体外培养大鼠原代VSMC,再将细胞分为空白对照组、miR-NC组、PDGF-BB+miR-145组及PDGF-BB组。CCK-8法检测各组细胞的增殖情况;实时RT-PCR方法检测PCNA、c-Jun及SM22a的表达水平;Western印迹方法检测ERK1/2和p-ERK1/2的表达、JNK和p-JNK的表达以及p38MAPK和p-p38MAPK的表达。结果 miR-145过表达后能够抑制PDGF诱导的大鼠原代VSMC增殖,并下调VSMC增殖相关基因PCNA、c-Jun的表达、上调分化相关基因SM22a的表达;PDGF-BB诱导VSMC后,ERK、JNK、p38MAPK的磷酸化水平均明显上调,而转染miR-145慢病毒后再加PDGF刺激,ERK、JNK、p38MAPK的磷酸化水平均明显下调。结论 miR-145能够抑制去分化型VSMC中的MAPK信号通路,进而抑制VSMC的增殖。     

p38MAPK反义寡核苷酸对鼠血管平滑肌细胞增殖的影响

目的：探讨丝裂素活化蛋白激酶p38(p38mitogen—activated protein kinase，p38MAPK)反义寡聚脱氧核苷酸(antisense oliodeoxynucleotide，AODN)对血管平滑肌细胞增殖的影响。方法：培养大鼠胸主动脉平滑肌细胞。通过脂质体帮染p38MAPK AODN到血管平滑肌细胞(VMSC)。另设p38MAPK正义寡聚脱氧核苷酸(SODN)对照组和空白对照组。用流式细胞仪检测细胞增殖。结果：AODN明显抑制血管紧张素I刺激的血管平滑肌细胞增殖(P＜0.05～＜0．01)，其抑制作用呈剂量依赖性。结论：p38MAPK反义寡聚脱氧核苷酸能抑制大鼠的VSMC增殖，提示VSMC增殖与p38信号途径有关。     

干扰趋化因子CC受体-2调控p38MAPK信号通路抑制人口腔黏膜上皮细胞系增殖的机制

目的探讨干扰趋化因子CC受体(CCR)2基因的表达对口腔黏膜上皮细胞增殖的影响及机制。方法 Western印迹检测CCR2在口腔黏膜下纤维性变(OSF)中的表达;用CCR2的siRNA转染人口腔黏膜上皮细胞系(HOMC)细胞,转染48 h后用Western印迹检测各组细胞中CCR2蛋白表达;细胞增殖与活性检测(CCK8)实验检测细胞增殖;Western印迹检测p38丝裂原活化蛋白激酶(p38MAPK)、磷酸化(p)-p38MAPK蛋白表达。结果 CCR2在OSF组织中的表达显著高于正常口腔黏膜组织(P<0.01);阴性对照(NC)-siRNA组CCR2蛋白表达、细胞存活率及p38MAPK、p-p38MAPK蛋白表达与对照组比较差异无统计学意义(P>0.05);CCR2-siRNA组CCR2蛋白表达、细胞存活率及p-p38MAPK蛋白表达均显著低于对照组(P<0.01)。结论 CCR2在OSF中高表达,抑制CCR2的表达可通过调控p38MAPK信号通路抑制HOMC的增殖。     

p38MAPK对轻度热应激大鼠脾脏巨噬细胞免疫功能的影响

目的: 探讨p38MAPK在Bip蛋白介导的体外轻度热应激大鼠巨噬细胞功能改变中的信号作用.方法: p38MAPK抑制剂预处理大鼠脾脏巨噬细胞,将细胞置于41℃恒温箱中,使细胞轻度热应激,1 h后恢复到37℃(抑制组),以未应激(对照组)和41℃热应激1 h后60 min巨噬细胞(应激组)为对照,分别检测3组巨噬细胞吞噬、杀伤、趋化功能,同时检测p38MAPK蛋白和Bip蛋白的表达.结果: p38MAPK抑制剂预处理大鼠脾脏巨噬细胞,与应激组比较,轻度热应激后巨噬细胞吞噬、趋化和杀伤活性明显降低(0.17±0.01 vs 0.74±0.03,33.32±3.55 vs 82.07±5.17,24.20%±2.39% vs 60.80%±4.02%,均P<0.01);应激组p38MAPK蛋白表达明显上调,p38MAPK抑制剂预处理后,抑制组p38 MAPK蛋白表达受到抑制,与应激组比较差异有显著性(p38/β-actin: 2.863±0.794 vs 4.752±1.386,P<0.01);Bip蛋白的表达(Bip/β-act in)也因p38MAPK抑制剂预处理而由应激组的1.270±0.535降至抑制组的1.028±1.061( P<0.05).结论: p38MAPK抑制剂可显著抑制轻度热应激大鼠巨噬细胞吞噬、趋化和杀伤功能以及p38MAPK和Bip蛋白的表达.     

右美托咪啶对大鼠肺IRI中p38MAPK信号通路及肺组织HMGB1表达的影响

目的研究右美托咪啶对大鼠肺缺血再灌注损伤(IRI)中p38丝裂素活化蛋白激酶(p38MAPK)及肺组织高迁移率族盒蛋白1(HMGB1)表达的影响,为分析右美托咪啶的肺保护作用奠定基础。方法将36只SD大鼠随机分为三组各12只:A组行假手术处理;B组建立大鼠急性肺IRI模型;C组在造模前以盐酸右美托咪啶预处理。手术3h后处死,取肺组织病理染色后观察组织病理学变化、行酶联免疫吸附试验检测肺组织髓过氧化酶(MPO)活性、行Western Blotting法检测肺组织磷酸化p38MAPK蛋白及HMGB1蛋白的表达。结果与A组比较,B组肺组织出现明显病理学改变,MPO活性明显更强,磷酸化p38MAPK蛋白及HMGB1蛋白表达水平亦明显增加,上述差异均有统计学意义(P0.05);与B组相比,C组上述各指标均明显下降,差异亦有统计学意义(P0.05)。磷酸化p38MAPK蛋白及HMGB1蛋白表达水平与肺组织病理损伤评分及MPO活性均呈显著正相关(P0.05)。结论肺IRI时可能出现p38MAPK信号通路异常活跃、肺组织HMGB1过表达,右美托咪啶则能有效抑制p38MAPK信号通路及肺组织HMGB1表达,这可能是其发挥肺保护作用的机制之一。     

姜黄素对血管平滑肌细胞增殖及cyclinD1和p21wafl/cipl蛋白表达的影响

目的 观察姜黄素(curcumin)抑制大鼠血管平滑肌细胞(VSMC)增殖和诱导细胞周期停滞的作用,以及对细胞周期蛋白cyclinDl,p21wafl/cipl表达的影响.方法 采用组织贴块法体外培养大鼠VSMC,MTT法测定姜黄素对VSMC增殖的抑制作用,流式细胞仪分析细胞周期分布,Western印迹法检测cyclinDl,p21wafl/cipl蛋白的变化.结果 MTT示不同浓度姜黄素(7.5～120 μmol/L)在24～72 h范围内,浓度和时间依赖性抑制VSMC增殖;30 μmol/L以上浓度姜黄素显著抑制增殖的VSMC细胞周期进程,使S及G2/M期减少(P<0.05),G0/G1期增加(P<0.05);30 μmol/L的姜黄素可抑制cyclinDl表达,促进p21wafl/cipl蛋白表达.结论 姜黄素具有明确的抑制VSMC增殖和细胞周期停滞的作用,其与cyclinDl,p21wafl/cipl蛋白变化有关.     

硫化氢通过p38MAPK信号通路对肝纤维化大鼠肝细胞增殖、凋亡的影响

目的:探讨硫化氢(hydrogen sulfide,H2S)对肝纤维化大鼠肝细胞增殖、凋亡的调节作用以及P-p38丝裂原活化蛋白激酶(p38mitogen-activated protein kinase,p38MAPK)蛋白的表达的影响.方法:用四氯化碳诱导肝纤维化SD大鼠模型,将提取的肝纤维化大鼠肝细胞分组:对照组、H2S组(对照组基础上加H2S的供体N a H S至最适浓度)、S B组(对照组基础上加SB203580至最适浓度)、SB+H2S组(对照组基础上加Na HS、SB203580至最适浓度).M T T法检测N a H S及S B203580对肝纤维化大鼠肝细胞的增殖、增殖抑制率的影响;Annexin V-FITC/PI双染流式细胞术检测肝纤维化大鼠肝细胞凋亡率;Western blot技术检测P-p38MAPK蛋白在各组中的表达水平.结果:与对照组相比,低浓度H2S(50μmol/L)促进肝纤维化大鼠肝细胞增殖明显(P=0.000),对肝纤维化大鼠肝细胞凋亡无影响;SB203580可抑制肝纤维化大鼠肝细胞的增殖,伴随药物浓度的升高细胞存活率降低(P=0.000),并诱导肝纤维化大鼠肝细胞凋亡(P=0.000);P-p38MAPK蛋白在各组中均有表达,H2S组表达水平高于对照组(P=0.000),SB组、SB+H2S组与对照组、H2S组相比,P-p38MAPK蛋白的表达量均减少(均P=0.000).结论:低浓度H2S对肝纤维化大鼠肝细胞凋亡无诱导作用,但能通过激活p38MAPK信号转导通路促进其细胞增殖.     

血管平滑肌细胞细胞周期素依赖性激酶抑制因子的调节

目的 阐明有丝分裂源激活的蛋白激酶(MAPK)和磷脂酶C(PLC)通路在调节细胞周期紊依赖性激酶抑制因子(CKI)p27，p21，p57蛋白表达量中的作用及其对血管平滑肌细胞(VSMC)增殖的影响。方法 分离SD大鼠主动脉中层平滑肌，贴壁法培养平滑肌细胞，无血清培养基培养静止后，分别加入血小板源性生长因子(PDGF)(20ng／m1)、PDGF PD98059(20ng／ml 20mmol／L)、PDGF 硫酸新霉紊(20ng／ml 10mmol／L)等刺激因素，以无血清培养基培养的VSMC作对照。在刺激后90min收集细胞，用免疫沉淀法检测MAPK(p44／p42)活性的改变。在刺激后6和24h收集细胞，用Western蛋白印迹法检测p27、p57和p21蛋白表达量。结果 PDGF刺激后，MAPK活性明显升高(较对照组增加46．6％)，同时VSMC明显增殖。刺激24h后，细胞增殖程度为对照组的1．43倍，p27蛋白的表达量显著下降至对照组的71％；p21和p57蛋白表达量却明显增加；加用PD98058(MAPK抑制剂)和新霉紊(PLC抑制剂)可明显抑制PDGF引起的上述改变，MAPK活性分别较PDGF。刺激组下降了46％和37％，p27蛋白表达量则分别为PDGF刺激组的1．77倍和1．49倍，细胞增殖程度分别降为后者的68％和65％。结论 MAPK(-14／42)变化的幅度是决定CKI表达量和VSMC增殖的关键因素。PLC通路在PDGF刺激VSMC增殖信号转导中同样起关键作用。     

p300/CBP相关因子对大鼠血管平滑肌细胞增殖的影响及机制研究

目的探讨p300/CBP相关因子(PCAF)对大鼠血管平滑肌细胞(VSMCs)增殖的作用及机制。方法组织块贴壁法获取大鼠原代VSMCs,使用Ad-PCAF RNAi腺病毒转染VSMCs抑制PCAF表达。选择1μg/mL的脂多糖(LPS)作为VSMCs增殖诱导剂。将实验分为6组:对照组(A组)、对照组+1μg/mL脂多糖刺激组(B组)、Ad-GFP腺病毒转染组(C组)、Ad-GFP腺病毒转染+1μg/mL脂多糖刺激组(D组)、Ad-PCAF RNAi腺病毒转染组(E组)、Ad-PCAF RNAi腺病毒转染+1μg/mL脂多糖刺激组(F组),使用CCK-8和流式细胞周期检测细胞增殖情况,各组中PCAF表达采用实时荧光定量聚合酶链式反应检测,蛋白免疫印迹法检测各组丝裂原活化蛋白激酶p38(p38 MAPK)、磷酸化丝裂原活化蛋白激酶p38(p-p38 MAPK)、丝氨酸/苏氨酸蛋白激酶(AKT)、磷酸化丝氨酸/苏氨酸蛋白激酶(p-AKT)表达水平。结果 1μg/mL脂多糖明显促进VSMCs增殖;同时,下调PCAF表达后,VSMCs增殖活性明显降低(P0.05)。流式细胞周期检测结果显示,脂多糖可促进VSMCs由S期向G2期转化,而下调PCAF后可明显抑制脂多糖的作用(P0.05)。同时,脂多糖诱导后VSMCs中PCAF mRNA表达水平明显增强;转染Ad-PCAF RNAi腺病毒后可明显降低VSMCs内PCAF mRNA的表达水平(P0.05)。此外,脂多糖诱导后VSMCs中p-p38 MAPK、p-AKT表达水平均明显升高(P0.05);而下调PCAF表达后,p-p38 MAPK、p-AKT表达均明显降低(P0.05)。结论下调PCAF的表达对VSMCs增殖具有明显的抑制作用,其机制可能与抑制细胞内p-p38 MAPK、p-AKT表达有关。     

山楂叶总黄酮对慢性脑缺血大鼠p38MAPK信号通路的影响

目的探讨山楂叶总黄酮(TFHL)对慢性脑缺血大鼠脑组织p38丝裂原活化蛋白激酶(MAPK)信号通路的影响。方法健康雄性SD大鼠,随机分为假手术组、模型组、TFHL组、银杏叶片组。采用永久性双侧颈总动脉结扎法制备慢性脑缺血模型。Morris水迷宫检测大鼠学习记忆能力;Western印迹法检测p38蛋白的表达。结果与模型组比较,TFHL可明显改善血管性痴呆大鼠学习记忆能力,缩短大鼠逃避潜伏期(P<0.05),增加大鼠穿越平台次数(P<0.05),抑制大鼠缺血脑组织p38蛋白的表达(P<0.05)。结论 TFHL对慢性脑缺血大鼠的神经功能具有保护作用,其机制可能与抑制p38蛋白表达,减轻脑神经元凋亡有关。     

Kallistatin stimulates vascular smooth muscle cell proliferation and migration in vitro and neointima formation in balloon-injured rat artery

Kallistatin, a serine proteinase inhibitor (serpin), is expressed in the endothelial and smooth muscle cells of blood vessels. The potential function of kallistatin in vascular biology was investigated by studying its role in the proliferation and migration of cultured primary aortic vascular smooth muscle cells (VSMCs) in vitro and in neointima formation in rat artery after balloon angioplasty in vivo. Exogenous kallistatin induced a >2-fold increase of VSMC proliferation and cell growth as measured by [(3)H]thymidine incorporation and cell counts and a 2.3-fold increase of cell migration in modified Boyden chambers. In balloon-injured vessels, endogenous kallistatin mRNA and protein levels increased up to 10-fold as determined by competitive polymerase chain reaction and by ELISA. Intense staining of kallistatin mRNA was identified in the proliferating VSMCs of balloon-injured arteries during cell migration from media to neointima by in situ hybridization histochemistry and immunohistochemistry. We observed an induction of kallistatin expression by platelet-derived growth factor (PDGF) and upregulation of p42/44 mitogen-activated protein kinase (MAPK) activity by kallistatin in cultured VSMCs. Conversely, adenovirus-mediated transfer of kallistatin antisense cDNA into cultured VSMCs inhibited PDGF-induced p42/44 MAPK activity and cell proliferation. Furthermore, local delivery of adenovirus carrying kallistatin antisense cDNA significantly downregulated kallistatin mRNA levels and attenuated neointima formation in balloon-injured rat arteries in vivo. These results indicate that kallistatin may play an important role in mediating PDGF-induced MAPK pathway on VSMC proliferation and in neointima formation after balloon angioplasty.     

雌激素膜快速效应对兔血管平滑肌细胞iNOS快速激活的影响

目的 探讨17β-雌二醇（17β-estradiol,E2）通过快速激活诱导型一氧化氮合成酶（iNOS）的活性,调节细胞外信号调节激酶（mitogen-activated protein kinase,p42/44 MAPK）表达抑制血管平滑肌细胞（vascular smooth muscle cells,VSMCs）增殖的机制。方法 在培养的VSMCs的基础上,采用放射免疫测定法(RIA)和Western blot检测E2预处理前后胎牛血清（fetal calf serum,FCS）对VSMCs中诱导型iNOS的活性及磷酸化p42/44 MAPK蛋白和p42/44 MAPK总蛋白表达的影响。采用比色法检测E2预处理后VSMCs中NO含量的变化。结果 E2预处理5 min,即可逆转FCS诱导的iNOS活性下降,此效应在E2预处理15 min时达到高峰,处理30 min后此效应减弱。基因转录抑制剂放线菌素D（actinomycin D,Act D）预处理后对此过程无影响,而三苯氧胺预处理可明显逆转E2诱导的iNOS活性增加。比色法检测上清液中NO的含量显示,E2预处理15 min,NO的含量明显增加。Western blot的结果显示,E2预处理15 min,可明显抑制磷酸化p42/44 MAPK蛋白的表达,而对总p42/44 MAPK蛋白的表达无影响。NO合酶(NOS)抑制剂L-硝基左旋精氨酸甲酯(L-NAME)预处理,可逆转E2诱导的磷酸化p42/44 MAPK的表达下降。结论 E2可通过快速激活iNOS增加NO释放,下调磷酸化p42/44 MAPK的活性,抑制VSMCs增殖。     

Nuclear accumulation of the AT1 receptor in a rat vascular smooth muscle cell line: effects upon signal transduction and cellular proliferation

The objective of the study was to identify the functional outcome of intracellular versus extracellular angiotensin II-AT(1) receptor interactions in vascular cells. Rat vascular smooth muscle cell line A10 was transfected, independently and concurrently, with plasmids encoding fluorescent fusion proteins of rat angiotensin II (pECFP/AII, encodes AII fused downstream of enhanced cyan fluorescent protein) and the rat AT(1a) receptor (pAT(1)R/EYFP, encodes the rat AT(1a) receptor fused upstream of enhanced yellow fluorescent protein). The AII fluorescent fusion protein possesses no secretory signal peptide and deconvolution microscopy established that is maintained within these cells predominantly in the nucleus. AT(1)R/EYFP was absent from the nucleus when expressed exclusively or in untreated cells but accumulated in the nucleus following exogenous AII treatment or when co-expressed with ECFP/AII. Furthermore, expression of ECFP/AII stimulated proliferation of A10 vascular smooth muscle cells (VSMCs) 1.6-fold (P < 0.05). Transfection of a control, pECFP/AII(C) (which encodes a scrambled AII peptide fused to ECFP) had no growth effect. In light of the intracellular growth effects of ECFP/AII, we sought to elucidate the underlying signaling pathways. We found that extracellular AII treatment of A10 cells activated cAMP response element-binding protein (CREB) as determined by one-hybrid assays and immunoblots. Expression of intracellular ECFP/AII similarly activated CREB. However, intracellular and extracellular AII activated CREB through different phosphorylation pathways. Exogenous AII treatment of A10 cells activated p38MAPK and ERK1/2 phosphorylation as determined by Western blot analyses and one-hybrid assays. The p38MAPK inhibitor, SB203580, and the ERK kinase inhibitor, PD98059 each partially inhibited exogenous AII-conferred CREB activation confirming that p38MAPK and ERK1/2 mediate CREB phosphorylation in this system. In contrast, expression of ECFP/AII (intracellular AII) in A10 VSMCs activated p38MAPK but not ERK1/2; inhibition of p38MAPK by SB203580 inhibited intracellular AII-induced CREB phosphorylation. In summary, extracellular AII stimulates at least one pathway common to intracellular AII. This common pathway, in the case of exogenous AII, likely reflects intracellular signaling following internalization of receptor-ligand complex. Extracellular AII also stimulates a unique pathway, apparently reflecting interaction with plasma membrane-associated AT(1)R.     

Chlamydia pneumoniae and chlamydial heat shock protein 60 stimulate proliferation of human vascular smooth muscle cells via toll-like receptor 4 and p44/p42 mitogen-activated protein kinase activation

An early component of atherogenesis is abnormal vascular smooth muscle cell (VSMC) proliferation. The presence of Chlamydia pneumoniae in many atherosclerotic lesions raises the possibility that this organism plays a causal role in atherogenesis. In this study, C pneumoniae elementary bodies (EBs) rapidly activated p44/p42 mitogen-activated protein kinases (MAPKs) and stimulated proliferation of VSMCs in vitro. Exposure of VSMCs derived from human saphenous vein to C pneumoniae EBs (3x10(7) inclusion forming units/mL) enhanced bromodeoxyuridine (BrdU) incorporation 12+/-3-fold. UV- and heat-inactivated C pneumoniae EBs also stimulated VSMC proliferation, indicating a role of direct stimulation by chlamydial antigens. However, the mitogenic activity of C pneumoniae was heat-labile, thus excluding a role of lipopolysaccharide. Chlamydial hsp60 (25 microg/mL) replicated the effect of C pneumoniae, stimulating BrdU incorporation 7+/-3-fold. Exposure to C pneumoniae or chlamydial hsp60 rapidly activated p44/p42 MAPK, within 5 to 10 minutes of exposure. In addition, PD98059 and U0126, which are two distinct inhibitors of upstream MAPK kinase 1/2 (MEK1/2), abolished the mitogenic effect of C pneumoniae and chlamydial hsp60. Toll-like receptors (TLRs) act as sensors for microbial antigens and can signal via the p44/p42 MAPK pathway. Human VSMCs were shown to express TLR4 mRNA and protein, and a TLR4 antagonist abolished chlamydial hsp60-induced VSMC proliferation and attenuated C pneumoniae-induced MAPK activation and VSMC proliferation. Together these results indicate that C pneumoniae and chlamydial hsp60 are potent inducers of human VSMC proliferation and that these effects are mediated, at least in part, by rapid TLR4-mediated activation of p44/p42 MAPK.     

JAK2/STAT3信号通路在阿托伐他汀抗大鼠血管平滑肌细胞增殖凋亡中的作用研究

目的探讨阿托伐他汀对大鼠血管平滑肌细胞(VSMCs)非调脂作用机制与转录信号传导子和激活子3(Stat3)信号传导通路的关系,明确其细胞内信号传导机制。方法分别将阿托伐他汀、酪氨酸激酶抑制剂AG490及二者联合作用于大鼠VSMCs,应用MTT法检测细胞增殖状态;流式细胞仪观察药物对细胞凋亡的影响,进一步用蛋白免疫印迹法(Western blot)检测药物作用前后 Stat3通路相关蛋白JAK2、STAT3、Cyclin D1、Bcl-2的表达及磷酸化活性。结果阿托伐他汀及AG490 都呈浓度依赖性方式抑制大鼠VSMCs增殖水平,促进其凋亡,二者联合应用则具协同作用。阿托伐他汀及AG490处理VSMCs后p-JAK2、p-Stat3、Cyclin D1、Bcl-2蛋白表达水平随作用时间延长而下降 (P<0．05),并且联合组蛋白表达水平较单独处理组相比有显著性差异(P<0．01)。结论阿托伐他汀对大鼠VSMCs非调脂作用机制与癌基因Star3信号通路高度相关,与酪氨酸激酶抑制剂AG490联合应用则具协同作用,可能通过作用于JAK2／Stat3下游靶基因Cyclin D1、Bcl-2影响其增殖及凋亡。     

信号转导子和激活子3通路调控大鼠血管平滑肌细胞增殖迁移机制研究

目的研究转录信号转导子和激活子3（stat3）通路与大鼠血管平滑肌细胞（VSMCs）增殖迁移的关系,明确VSMCs增殖的信号转导过程。方法应用脂质体转染Stat3反义寡核苷酸作用于大鼠VSMCs,应用酶联免疫法（ELISA）检测Stat3水平变化及MTF法检测细胞增殖状态,蛋白免疫印迹法（Western blot）检测stat3、磷酸化stat3及其靶基因产物Cyclin D1、Bcl—XL的表达。结果转染Stat3反义寡核苷酸后,大鼠VSMCs中Stat3水平明显下降（P〈0．01）,同时其增殖水平降低,而相应空白对照组、脂质体组、转染正义寡核苷酸组变化不明显。转染Stat3反义寡核苷酸的VSMCs中stat3、p-Stat 3、Cyclin D1蛋白表达水平随作用时间延长而下降（P〈0．01）,而Bcl—xL水平无明显变化。结论癌基因stat3信号通路与大鼠VSMCs增殖高度相关,可能通过其下游靶基因Cyclin D1影响其增殖,而阻断此通路则可抑制VSMCs的增殖。     

缬沙坦对血管平滑肌细胞迁移及丝裂原活化蛋白激酶的影响

目的观察缬沙坦（Val）对血管紧张素Ⅱ（AngⅡ）刺激下大鼠血管平滑肌细胞（VsMC）迁移及磷酸化42／44丝裂原活化蛋白激酶（p42／44MAPK）表达的影响。方法组织贴块法培养大鼠胸主动脉平滑肌细胞,tran-sweⅡ小室检测细胞的迁移能力,免疫印迹法检测p42／44MAPK蛋白表达的水平。结果（1）AngⅡ能明显促进VSMC迁移,该作用可被Val和MAPK激酶的特异性抑制剂PD98059所抑制。（2）AngⅡ刺激VSMC5min时,p42／44MAPK的表达量最大,该作用可被Val和PD98059所抑制。（3）Val单独作用于VSMC时,对细胞的迁移及p42／44MAPK的表达均无明显影响。结论Val抑制AngⅡ诱导的VSMC迁移与其抑制AngⅡ诱导的p42／44MAPK的表达相关。