蛋白激酶Cα、β及Rho激酶对宫缩乏力性产后出血产妇子宫平滑肌收缩功能影响研究

目的探讨蛋白激酶C(PKC)α和PKCβ特异性抑制剂Go6976、PKCβ特异性抑制剂LY333531和Rho激酶特异性抑制剂Y-27632对宫缩乏力性产后出血产妇子宫平滑肌收缩的抑制作用。方法 2010年11月至2011年12月于福建省妇幼保健院选择剖宫产分娩的初产妇60例,分宫缩乏力性产后出血组(病例组,n=30)和宫缩良好无产后出血组(对照组,n=30)。采用肌条等长张力测定方法检测两组产妇子宫下段平滑肌分别在Go6976、LY333531和Y-27632作用下自发性收缩功能变化及病例组在LY333531、Y-27632作用下梯度浓度缩宫素诱导的收缩潜能变化。结果 Go6976、LY333531和Y-27632均显著抑制两组产妇子宫平滑肌收缩幅度及活动力(均P0.05);对病例组抑制作用均小于对照组(均P0.05),而对两组的收缩频率无影响(P0.05)。LY333531和Y-27632均可显著降低病例组收缩潜能(P0.05)。结论 PKCα、PKCβ和Rho激酶影响子宫平滑肌收缩,可能参与宫缩乏力性产后出血的发生。     

The Inhibitory Effect of Shikonin on the Agonist-Induced Regulation of Vascular Contractility

Shikonin, a natural flavonoid found in the roots of Lithospermum erythrorhizon, has been shown to possess many biological functions. The present study was undertaken to investigate the influence of shikonin on vascular smooth muscle contractility and to determine the mechanism involved. Denuded aortic rings from male rats were used and isometric contractions were recorded and combined with molecular experiments. Shikonin significantly relaxed fluoride-, thromboxane A₂- or phorbol ester-induced vascular contraction suggesting as a possible anti-hypertensive on the agonist-induced vascular contraction regardless of endothelial nitric oxide synthesis. Furthermore, shikonin significantly inhibited fluoride-induced increases in pMYPT1 levels and phorbol ester-induced increases in pERK1/2 levels suggesting the mechanism involving the inhibition of Rho-kinase activity and the subsequent phosphorylation of MYPT1 and the inhibition of MEK activity and the subsequent phosphorylation of ERK1/2. This study provides evidence regarding the mechanism underlying the relaxation effect of shikonin on agonist-induced vascular contraction regardless of endothelial function.     

Administration of the Rho-Kinase Inhibitor Fasudil Before Ischemia or just After Reperfusion,but not 30 min After Reperfusion,Protects the Stunned Myocardium in Swine

OBJECTIVES: We assessed the effect of administration time for fasudil treatment of the stunned myocardium in 40 anesthetized open chest swine. MATERIALS AND METHODS: All swine were subjected to 12 min ischemia followed by reperfusion to generate stunned myocardium. Group A (n = 11) received saline in place of fasudil both before ischemia and after reperfusion. Group B (n = 10) received 30 min intravenous fasudil at a rate of 13 mug/kg/min starting 45 min before ischemia and received saline after reperfusion. Groups C (n = 10) and D (n = 9) received saline before ischemia, and received fasudil at a rate of 13 mug kg(-1) min(-1) starting just before reperfusion in group C and 30 min after reperfusion in group D. In both groups, treatment lasted 30 min. Myocardial contractility was assessed by percent segment shortening (%SS). RESULTS AND DISCUSSION: Three swine in group A, 2 swine in each of groups B and C, and one swine in group D had ventricular fibrillation or tachycardia after reperfusion and were excluded from further analysis. The changes of %SS from baseline at 90 min after reperfusion in groups B and C were 68 +/- 8% and 75 +/- 8%, respectively, which were significantly higher than in group A or D (47 +/- 10% or 43 +/- 8%). CONCLUSION: We conclude that fasudil administered before ischemia or just after reperfusion, but not 30 min after reperfusion, protects the stunned myocardium.     

白介素-1β诱导小型猪冠状动脉内膜增殖时Rho激酶表达与雷帕霉素干预的研究

目的通过小型猪模型观察白介素-1β(IL-1β)诱导冠状动脉内膜增殖时Rho激酶和p27mRNA表达的变化及雷帕霉素干预的作用,探讨Rho激酶表达对冠状动脉狭窄的作用及可能机制。方法通过开胸手术分离小型猪冠状动脉左前降支(LAD)及回旋支(LCX)近端,包裹IL-1β。2周后行冠状动脉造影,然后取标本做病理学检查,并用RT-PCR法测血管壁组织Rho激酶和p27mRNA表达。结果正常冠状动脉血管壁可见Rho激酶mRNA表达及较高水平的p27mRNA表达;用一定量IL-1β诱导冠状动脉内膜增殖,可迅速造成血管管腔狭窄。在此过程中Rho激酶mRNA表达增加3倍以上(30.80±4.10对128.20±15.89),而p27mRNA的表达则明显减弱。雷帕霉素抑制小型猪血管内膜增殖,减少炎细胞浸润,还能减少Rho激酶mRNA表达,并增强p27mRNA的表达。结论Rho激酶在炎症因子诱导的冠状动脉狭窄过程中表达明显增强,通过向下调节p27的活性调控血管内膜的增殖。雷帕霉素通过增强p27mRNA的表达,而减弱Rho激酶对p27的调节,这可能是雷帕霉素抑制血管内膜增殖的新机制。     

Rho/Rho激酶信号通路与冠心病

Rho/Rho激酶信号通路是体内普通存在的一条信号转导通路,它通过调节细胞内肌动蛋白骨架的聚合状态参与多种细胞功能,包括细胞收缩、迁移、黏附、增殖、凋亡及基因表达等。Rho/Rho激酶信号通路的异常激活在冠心病的发病机制和病理生理中发挥了重要作用,对此信号转导通路的研究可以为冠心病的预防和治疗提供新的靶点。现就Rho/Rho激酶信号通路的特征及其与冠心病的关系作一综述。     

Rho激酶与支架内再狭窄

目的 支架内再狭窄与血管平滑肌分化、迁移,细胞外基质的过度增值所致新生内膜增生密切相关.Rho激酶参与支架置入引起的新生内膜增生的调节.长期抑制Rho激酶的表达可阻止新生内膜的增生,可能成为防止支架内再狭窄的一种方法.     

Leads for the treatment of pulmonary hypertension

Background: Knowledge of the causative reasons for pulmonary arterial hypertension, a major category of pulmonary hypertension, has expanded dramatically over the past 10 years. This has led to heightened research across a range of potential new mechanistic approaches and resulted in the identification of further treatment options, together with several promising leads and prototypes. Objective: This review aims to summarise and assess the most relevant research fields, covering key publications and recent patent literature. Methods: Searching revealed in excess of 700 patents claiming uses that relate to pulmonary hypertension. These patents were filtered into key therapeutic approaches based on pharmacological reviews of the pulmonary arterial hypertension field. Results/conclusions: Endothelin antagonists and phosphodiesterase 5 inhibitors have emerged as recently approved treatment options and are proving extremely beneficial. Further new mechanistic approaches have yielded promising leads, some of which have the potential to be disease modifying; notably, tyrosine kinase inhibitors, soluble guanylate cyclase agonists, Rho-kinase inhibitors, vasoactive intestinal peptide analogues and 5-HT antagonists.     

Endothelium-Independent Effect of Fisetin on the Agonist-Induced Regulation of Vascular Contractility

Fisetin, a natural flavonoid found in a variety of vegetables and fruits, has been shown to possess many biological functions. The present study was undertaken to investigate the influence of fisetin on vascular smooth muscle contractility and to determine the mechanism involved. Denuded aortic rings from male rats were used and isometric contractions were recorded and combined with molecular experiments. Fisetin significantly relaxed fluoride-, thromboxane A₂- or phorbol ester-induced vascular contraction suggesting as a possible anti-hypertensive on the agonist-induced vascular contraction regardless of endothelial nitric oxide synthesis. Furthermore, fisetin significantly inhibited fluoride-induced increases in pMYPT1 levels and phorbol ester-induced increases in pERK1/2 levels suggesting the mechanism involving the inhibition of Rho-kinase activity and the subsequent phosphorylation of MYPT1 and MEK activity and the subsequent phosphorylation of ERK1/2. This study provides evidence regarding the mechanism underlying the relaxation effect of fisetin on agonist-induced vascular contraction regardless of endothelial function.     

Rho激酶抑制剂在急性冠脉综合征介入治疗中的应用价值

目的应用SYNTAX评分,观察Rho激酶抑制剂法舒地尔对不同评分组的急性冠脉综合征患者介入术后左心功能、一年支架内再狭窄(ISR)和主要心脏不良事件(MACE)的影响。方法入选急性冠脉综合征患者238例,均经冠脉造影证实存在单支以上的病变,均拒绝行冠脉搭桥手术,按随机数字表法分为治疗组(法舒地尔,n=102)和对照组(n=136),根据SYNTAX评分,每组再分为评分低值亚组(0~22分)、评分值中等亚组(23~32分)和评分值高亚组(≥33分),正规冠心病二级预防治疗,并行冠脉造影和支架植入治疗,随访一年,对比观察2组以及2组的亚组间在术中慢血流或无复流、急性或亚急性支架血栓、左心功能、一年内支架内再狭窄率和MACE事件的差异。结果治疗组和对照组观察的各项指标无显著差异(P0.05),2组对应的评分低值组亚组间以及评分中等值亚组间观察的各项指标无显著差异(P0.05);与对照组的评分高值亚组相比,治疗组的评分高值亚组术中发生慢血流或无复流的比例、1年支架内再狭窄比例和MACE事件的发生率显著降低(P0.05),3个月的左心功能明显改善(P0.05)。结论 Rho激酶抑制剂可显著减少SYNTAX评分高值(≥33分)的ACS患者介入术中慢血流或无复流的发生,改善左心功能,并降低1年内支架内再狭窄率和MACE事件的发生率。     

The Rho-kinase inhibitor inhibits proliferation and metastasis of small cell lung cancer

The purpose of this study was to investigate the effects of Rho-kinase inhibitor on the growth, proliferation, apoptosis, adhesion, invasion and migration of NCI-H446 small cell lung cancer cells and to explore the underlying molecular mechanisms involved in this process. After treatment to NCI-H446 small cell lung cancer cells with Fasudil, a Rho-kinase inhibitor, cell biological behaviors were observed. Matrix metalloproteinase activity and Western blot assay were used to evaluate underlying molecular mechanisms. The IC50 of Fasudil to NCI-H446 small cell lung cancer cells was approximately 0.86 mg/ml (95% confidence limits: 0.65–1.17 mg/ml). After treatment with 0.75 mg/ml Fasudil, the ability of NCI-H446 small cell lung cancer cells, including growth, proliferation, adhesion, migration, and invasion were decreased, while their apoptosis was increased significantly. On the molecular level, the total amounts of active MMP2 and MMP9 were decreased about 20.5% (P < 0.05) and 57.5% (P < 0.01) respectively. Myosin phosphatase targeting subunit 1 phosphorylation (P-MYPT1) was reduced by 27.9% (P < 0.05). The activation of caspase-3, and PARP cleavage in experimental group were significantly higher than those in normal control group (P < 0.01). Meanwhile, treatment with Fasudil led to a remarkable reduction of the phosphorylated STAT3 (P-STAT3) (P < 0.01). Taken together, our findings show that Fasudil prevents the growth, metastasis and induces apoptosis of NCI-H446 small cell lung cancer cells by inhibiting the Rho/Rho-kinase pathway. Changes in MMP2, MMP9, P-MYPT1, caspase-3, PARP cleavage and P-STAT3 may be one of its molecular mechanisms.