Fasudil inhibits tissue factor and plasminogen activator Inhibitor-1 secretion by peripheral blood mononuclear cells in CAPD patients

Disturbances in hemostasis are common complications of kidney diseases and correlate well with cardiovascular mortality. Little is known about the effects of fasudil on tissue factor (TF) and plasminogen activator inhibitor-1 (PAI-1) expression in peripheral blood mononuclear cells (PBMCs) in CAPD patients. PBMCs were isolated from 13 individuals with CAPD and 13 healthy subjects. After 4?h of incubation with or without LPS (10?ng/mL), TF and PAI-1 mRNA of PBMCs were detected by RT-PCR. The levels of TF and PAI-1 in culture supernatants of PBMCs were determined by ELISA. Compared with healthy controls, CAPD patients had increased TF, PAI-1 protein and mRNA expression by PBMCs at baseline and after stimulated by LPS (10?ng/mL) [p?<?0.001]. The fasudil treatment resulted in a significant effect in decreasing TF and PAI-1 [p?<?0.05] synthesis in PBMCs. TF and PAI-1 mRNA expression and activities in PBMCs were increased in CAPD patients. Fasudil reduced LPS-mediated TF and PAI-1 expression and activity in PBMCs. These effects may partially be relevant to the clinical benefits of fasudil in the treatment of CAPD patients.     

法舒地尔对SH-SY5Y细胞氧糖剥夺后突触损伤的影响

目的:探讨法舒地尔(Fasudil)对人神经母细胞瘤细胞(SH-SY5Y)氧糖剥夺(OGD)后突触损伤的影响。方法:培养SH-SY5Y细胞,分为空白对照组、OGD组、OGD+法舒地尔组,各3皿。相差光学显微镜下观察细胞突触损伤及修复的细胞形态;Western-Blot检测ROCKⅡ、磷酸化肌球蛋白磷酸酶(p-MYPT1)、突触后致密物-95(PSD-95)、突触素(Synaptophysin)等蛋白的表达情况。结果:形态学显示法舒地尔可有效修复OGD诱导的神经突触损伤。OGD组ROCKⅡ及p-MYPT1的表达明显高于空白对照组(P0.05),突触素及PSD-95的表达明显低于空白对照组(P0.01)。OGD+法舒地尔组ROCKⅡ及p-MYPT1表达水平低于OGD组(P0.05),而与对照组差异无统计学意义(P0.05);突触素和PSD-95表达水平高于OGD组(P0.01);PSD-95表达水平高于空白对照组(P0.05),突触素的表达与空白对照组差异无统计学意义(P0.05),结论:法舒地尔可有效修复OGD诱导的神经突触损伤,增加PSD-95、突触素的表达,抑制ROCKⅡ及p-MYPT1的表达。     

银杏叶片联合法舒地尔治疗冠心病慢性心力衰竭的临床研究

目的探讨银杏叶片联合盐酸法舒地尔注射液治疗冠心病慢性心力衰竭的临床疗效。方法选取2016年6月—2017年9月凉山彝族自治州第一人民医院收治的冠心病合并慢性心力衰竭患者98例为研究对象,患者随机被分为对照组和治疗组,每组各49例。对照组静脉滴注盐酸法舒地尔注射液,30 mg加入到5%葡萄糖溶液100 mL中,2次/d,10 d为1个疗程,每个月治疗1个疗程。治疗组在对照组基础上口服银杏叶片,1片/次,3次/d。两组患者均连续治疗3个月。观察两组的临床疗效,比较两组的6 min步行距离、心功能分级、超声心动图指标、血清生化指标。结果治疗后,对照组和治疗组的总有效率分别为77.6%、91.8%,两组比较差异具有统计学意义(P0.05)。治疗后,两组6 min步行距离均明显增加,心功能分级均明显改善,同组治疗前后比较差异具有统计学意义(P0.05);且治疗组这些观察指标的改善程度明显优于对照组,两组比较差异具有统计学意义(P0.05)。治疗后,两组左心室舒张末期内径(LVEDD)、左心室收缩末期内径(LVESD)均显著降低,左室射血分数(LVEF)、每搏量(SV)均显著升高,同组治疗前后比较差异具有统计学意义(P0.05);且治疗组这些观察指标的改善程度明显优于对照组,两组比较差异具有统计学意义(P0.05)。治疗后,两组血清脑钠肽(BNP)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)水平均显著降低,同组治疗前后比较差异具有统计学意义(P0.05);且治疗组这些观察指标明显低于对照组,两组比较差异具有统计学意义(P0.05)。结论银杏叶片联合盐酸法舒地尔注射液治疗冠心病慢性心力衰竭具有较好的临床疗效,可改善患者的心功能,降低炎性指标,提高运动能力,安全性较好,具有一定的临床推广应用价值。     

法舒地尔联合纳洛酮治疗外伤性脑梗死的疗效观察

目的探讨法舒地尔联合纳洛酮治疗外伤性脑梗死的临床疗效。方法选择2013年1月—2018年1月山东省立第三医院收治的外伤性脑梗死患者110例,随机分为对照组和治疗组,每组各55例。对照组静脉滴注盐酸纳洛酮注射液,0.8mg加入250 mL生理盐水,1次/d。治疗组在对照组基础上静脉滴注盐酸法舒地尔注射液,30 mg加入250 mL生理盐水,1次/d。两组患者连续治疗14 d。观察两组患者临床疗效,同时比较治疗前后两组患者神经功能和运动功能FM评分。结果治疗后,对照组和治疗组临床有效率分别为76.36%和92.73%,两组比较差异具有统计学意义(P0.05)。治疗后,两组患者Barthel指数和欧洲版神经功能缺损评分(ESS)评分均显著升高(P0.05),美国国立卫生研究卒中量表(NIHSS)显著降低(P0.05),且治疗组神经功能评分明显优于对照组(P0.05)。治疗后,两组患者上肢和下肢及总评分均显著升高(P0.05),且治疗组患者治疗后运动功能FM评分明显高于对照组(P0.05)。结论法舒地尔联合纳洛酮治疗外伤性脑梗死能够显著提高临床疗效,具有一定的临床推广应用价值。     

前列地尔与法舒地尔治疗急性脑梗死的对比研究

目的比较前列地尔与法舒地尔治疗急性脑梗死的临床疗效。方法选取2013年1月—2015年7月榆林市第一医院收治的急性脑梗死患者92例,随机分为对照组(44例)和治疗组(48例)。对照组静脉滴注盐酸法舒地尔注射液,60 mg加入到生理盐水250 m L中,1次/d。治疗组静脉滴注前列地尔注射液,20μg加入到生理盐水250 m L中,1次/d。两组患者均连续治疗2周。观察两组的临床疗效,比较两组NISS评分、正五聚体蛋白3(PTX-3)及颈动脉粥样斑块的变化。结果治疗后,对照组和治疗组的总有效率分别为77.3%、93.8%,两组比较差异有统计学意义(P0.05)。治疗后,两组NISS评分均显著降低,同组治疗前后比较差异有统计学意义(P0.05);且治疗组NISS评分的下降程度明显优于对照组,两组比较差异具有统计学意义(P0.05)。治疗后,两组PTX-3均显著降低,同组治疗前后比较差异有统计学意义(P0.05);且治疗组PTX-3的下降程度明显优于对照组,两组比较差异具有统计学意义(P0.05)。治疗后,两组斑块形成率显著下降,而稳定斑块比例上升,治疗组同组治疗前后比较差异有统计学意义(P0.05)。对照组和治疗组斑块形成率分别为35.4%、56.8%,稳定斑块所占比例分别为52.0%、94.1%,两组比较差异具有统计学意义(P0.05)。结论前列地尔治疗急性脑梗死具有较好的临床疗效,可改善临床症状,降低PTX-3,提高斑块稳定性,安全性较好,具有一定的临床推广应用价值。     

法舒地尔对人肺动脉平滑肌细胞增殖抑制作用研究

目的:探讨法舒地尔对血小板源性生长因子(platelet-derived growth factor,PDGF)诱导的人肺动脉平滑肌细胞(human pulmonary artery smooth muscle cell,HPASMC)增殖的抑制作用及机制。方法:以体外培养的HPASMC为研究对象,PDGF-BB诱导增殖,法舒地尔进行预处理,MTT法检测细胞增殖;流式细胞仪检测细胞周期;Western blot检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)和p27kip1蛋白的表达。结果:细胞培养24 h后与空白组比较,PDGF组HPASMC细胞增殖比例,S期细胞比例和PCNA蛋白表达明显增加,p27kip1蛋白表达则明显降低;用法舒地尔预处理后,与PDGF组比较,细胞增殖比例,S期细胞比例和PCNA蛋白表达明显下降,p27kip1蛋白表达则明显增加。结论:法舒地尔可通过上调p27kip1蛋白表达来抑制PDGF诱导的HPASMC增殖和细胞周期进程。     

法舒地尔和RhoA沉默对神经干细胞增殖的影响

背景：Rho及其相关分子在神经轴突生长、分化、延伸及突触形成中起重要作用,阻断和抑制RhoA/ROCK通路可促进神经干细胞的增殖与生长。 目的：观察Rho激酶抑制剂法舒地尔和RNAi介导的RhoA基因沉默对大鼠神经干细胞增殖的影响。 方法：体外培养Wistar胎鼠神经干细胞,分6组干预：空白对照组,5,10,15,20 μmol/L 法舒地尔组,siRNA 沉默RhoA基因组。干预后第3天,采用RT-PCR,Western blot检测各组神经干细胞RhoA基因及蛋白的表达。应用MTT比色法观察神经干细胞增殖情况;采用流式细胞术测定神经干细胞周期分布的变化。 结果与结论：15,20 μmol/L 法舒地尔组、siRNA 沉默RhoA基因组神经干细胞RhoA基因及蛋白表达量较5,10 μmol/L 法舒地尔组、空白对照组明显降低(P < 0.05),细胞的生长速度较5,10 μmol/L 法舒地尔组、空白对照组明显增快(P < 0.05),细胞周期G0/G1期减少(P < 0.05),S期细胞数增多(P < 0.05)。当法舒地尔浓度增加到20 μmol/L时对细胞的作用并非随浓度的增加而增强,与15 μmol/L组的差异无显著性意义(P > 0.05)。15,20 μmol/L 法舒地尔组与siRNA 沉默RhoA基因组相比差异无显著性意义(P > 0.05)。说明Rho激酶抑制剂法舒地尔和RNAi介导的RhoA基因沉默在体外均能促进神经干细胞增殖,法舒地尔最佳作用浓度为15 μmol/L。     

Fasudil protects cultured N1E-115 cells against lysophosphatidic acid-induced neurite retraction through inhibition of Rho-kinase

The aim of this study was to investigate the possible effects of the Rho-kinase inhibitor, fasudil, on the lysophosphatidic acid (LPA)-induced neurite retraction in N1E-115 cells. In cultured N1E-115 cells, LPA produced a marked increase in the population of rounded cells. Fasudil or hydroxyfasudil, an active metabolite of fasudil, blocked cell rounding in a concentration-dependent manner at levels between 1 and 10 μM, with IC₅₀ values of 1.7 or 1.6 μM, respectively. Fasudil or hydroxyfasudil concentration-dependently inhibited phosphorylation of the myosin binding subunit of myosin phosphatase in N1E-115 cells. These results indicate that Rho-kinase is essential for LPA-induced neurite retraction in N1E-115 cells and that inactivation of Rho-kinase by a Rho-kinase inhibitor, such as fasudil, eliminates cell rounding and promotes neurite outgrowth, thus improving neurological function in the brain damage.     

法舒地尔在肝癌血管介入治疗中的应用

目的 探讨解除肝癌介入化疗栓塞过程中血管痉挛的方法.方法 肝癌栓塞化疗过程中,痉挛血管近侧局部经导管推注法舒地尔2.5 mg,然后行DSA造影确定血管扩张情况,同时观察并记录其不良反应.结果 30例次血管痉挛给予法舒地尔后全部解除,顺利完成超选择插管并给予足量超液化碘油充填,患者无特殊不适主诉.结论 导管内推注法舒地尔...     

法舒地尔对肺血管重构的肺动脉高压大鼠的影响

目的 观察法舒地尔对发生肺血管重构的肺动脉高压(PAH)大鼠的影响.方法 将32只雄性Wistar大鼠随机均分为4组:对照3周组,模型3、6周组及治疗组;予野百合碱建模;治疗组于3周后予法舒地尔腹腔内注射.于3、6周末测定各组大鼠右心室收缩压(RVSP)、内皮素-1(ET-1)、N端脑钠素前体(NT-proBNP)、肺小动脉病理等.结果 (1)造模后RVSP随时间升高(31.2±7.6)、(43.3±8.3)、(56.9±6.9)mm Hg(1 mm Hg=0.133 kPa),治疗组(47.3±6.3)mmHg较模型6周组明显降低.(2)ET-1、NT-proBNP与RVSP呈正相关(r=0.721、0.454).(3)病理:造模后肺动脉平滑肌增生,管壁狭窄,至6周时近于闭塞,治疗组则明显好转.结论 对于已发生肺血管重构的PAH大鼠,法舒地尔能阻止肺动脉高压进展,ET-1、NT-proBNP可成为较理想的评估指标.

Abstract：

Objective To investigate the effects of Rho kinase inhibitor fasudil on monocrotaline (MCT)-induced pulmonary arterial hypertension (PAH) with vascular remodeling in rats. Methods Thirty-two male Wistar rats were randomly and equally divided into control group ( C3 ), MCT-PAH 3- and 6-week model groups (H3, H6 ) and fasudil-treated group (F6). The right ventricle systolic pressure( RVSP), right ventricle/left ventricle and septum[RV/( LV + S)], plasma endothelin-1 and NT-proBNP were measured; small pulmonary arterial morphometry, ratio of vascular wall thickness to vascular external diameter (WT) and ratio of vascular wall area to the total area (WA) were also observed. Results ( 1 )The RVSP was highest in H6 group, followed by F6 group, H3 group, and C3 group[(56. 9 ±6. 9), (47. 3±6.3), (43.3±8.3) and (31.2±7.6) mm Hg (1 mm Hg=0. 133 kPa)]. But no significant difference was found between H3 and F6 groups; (2) The levels of plasma ET-1 and NT-proBNP were positively associated with RVSP, RV/(LV +S), WT and WA (r for ET-1 was 0.721, 0.607, 0.652, 0.707,P＜0.01; r for NT-proBNP was 0. 454, 0. 330, 0. 365, 0. 398,P ＜0. 05); (3) Pathematological assessment showed that the thickness and muscularization of small pulmonary artery were increased in H3 group as compared with C3 group, most significantly in H6 group, and those were significantly decreased in F6 group as compared with H6 group. Conclusion Fasudil can improve the MCT-induced PAH with vascular remodeling in rats. The plasma ET-1 and NT-proBNP could be used as good makers to evaluate the severity of PAH.