Experimental Studies on Preventive Effects of API_（0134） on Vessel Reocclusion after Thrombolysis

ＥｘｐｅｒｉｍｅｎｔａｌＳｔｕｄｉｅｓｏｎＰｒｅｖｅｎｔｉｖｅＥｆｆｅｃｔｓｏｆＡＰＩ_（０１３４）ｏｎ　ＶｅｓｓｅｌＲｅｏｃｃｌｕｓｉｏｎａｆｔｅｒＴｈｒｏｍｂｏｌｙｓｉｓＸＩＯＮＧＹｉ－ｌｉ（熊一力）；ＺＨＡＯＨｕａ－ｙｕｅ（赵华月）（Ｄｅｐａｒ?..     

Protective effects of API0134 on myocardial ischemia and reperfusion injury

Summary Previous studies have demonstrated that a crude extract from Chinese medicinal herbAndrographis Paniculata Nees (APN) could prevent myocardial ischemia and reperfusion injury. A refined extract API₀₁₃₄ was studied further. LAD was ligated for 90 min in 20 dogs and then reperfused for 120 min. The animals were randomly divided into 2 groups, API₀₁₃₄ treated group(n = 10), 45 min after ischemia receiving a slow i. v. bolus of 1 mg/kg and then an infusion of 80 μg D kg⁻¹/min for 60 min and control group (n=10) which was given only 5% glucose in saline. Result showed that the hemodynamics in API₀₁₃₄ treated group showed better effects of preventing the increase of the LVEDP and maintaining relatively normal CO as compared with control group. Ischemic ECGs were significantly milder. Malignant arrhythmia did not appear in API₀₁₃₄ treated group. After reperfusion, the infarct size was smaller (5. 06±2. 67% vs 10. 45±3. 11%,P < 0. 01), the damages found in myocardial ultrastructure were significantly milder. It is concluded that API₀₁₃₄ may protect the myocardium from ischemic reperfusion injury.     

OxLDL-induced cytotoxicity and its inhibition by API0134 in cultured porcine aortic endothelial cells

Summary Protective effects of API₀₁₃₄ on endothelial cells (EC) damaged by oxidatively modified low density lipoprotein (oxLDL) were studied. The results showed that the content of endothelia (ET) and malondialdehyde (MDA) in the media of porcine aortic EC incubated with oxLDL were increased and the cGMP was decreased significantly, and the activity of Superoxide dismutase (SOD) was inhibited. The effect of cytotoxicity of oxLDL can be eliminated by API₀₁₃₄. These results suggest that API₀₁₃₄ may protect EC against damages elicited by oxLDL. This project is supported by the National Natural Science Foundation of China (No. 39470859)     

心肌缺血预适应抗血栓形成作用的实验研究

观察缺血预适应对大白兔血小板聚集，血栓形成，GMP-140及PCI2的影响。将24只大白兔随机分为2组，实验组心脏冠脉左前降支（LAD）经过5min缺血，10min再灌注后持续缺血60min，3h再灌注；对照组直接造成LAD60min缺血，3h再灌注。比浊法测定血小板聚集活性，放免法测定血浆PGI2水平以及GMP-140分子数。另24只大白兔血小板聚集活性降低（ADP及AA诱导），血栓湿重降低，GMP-140分子数低于对照组，PGI2有升高趋势，差异有统计学意义。提示：缺血预适应可以抑制大白兔血小板的活化和聚集，减少血栓的形成。     

毛冬青皂苷B3的抗血栓作用研究

目的 从毛冬青根中分离、鉴定毛冬青皂苷B₃,研究其抗血栓作用。方法 色谱法、重结晶法分离、精制毛冬青皂苷B₃,薄层色谱对照及波谱分析法对毛冬青皂苷B₃进行结构鉴定。采用体内血栓形成试验、体外血凝块溶解试验及二磷酸腺苷（ADP）诱导血小板聚集试验,观察毛冬青皂苷B₃的抗血栓作用。结果 体内抗血栓试验显示,毛冬青皂苷B₃对胶原蛋白-肾上腺素诱发的血栓形成所致小鼠的偏瘫和死亡及FeCl₃诱导的大鼠腹主动脉血栓形成均有明显的抑制作用;在体外实验中,对家兔血凝块无明显的溶解作用,对ADP诱导的离体家兔血小板聚集也无明显的抑制作用。结论 毛冬青皂苷B₃可能为毛冬青抗血栓作用的物质基础之一,其体内抗血栓形成的作用可能与其促进血凝块溶解和抗ADP诱导的血小板聚集无关。     

Clinical studies on treatment of acute cerebral infarction with Xueshuantong injection

Objective: To observe the effect of Xueshuantong injection (XST, with its ingredient as Notoginseng saponin, on acute cerebral infarction (ACI) and on blood coagulation and fibrinolysis, so as to comprehensively analyse the mechanism of XST.Methods: Fifty ACI patients were randomly divided into 2 groups, and XST group (30 patients) was treated with XST, and the control group (20 patients) given low molecular dextrose, as well as low molecular heparin calcium. The course of treatment for both groups was 15 days. The changes of effective rate, score of neurologic impairment, tissuetype plasminogen activator (tPA), inhibitor of plasminogen activator (PAI), D-D dimmer, antithrombinIII (AT-III), and fibrinogen (Fbg) were all observed.Results: The total effective rate of XST group was 73.33%, that of the control group 65. 00%. After the therapy, plasma level of tPA, ratio of tPA/PAI, and AT-III content were increased obviously, while the plasma level of PAI and D-D dimmer were decreased significantly (all P<0.01). But there was only insignificant difference between the 2 groups (P> 0.05).Conclusion: XST injection could be effective to ACI, the mechanism of which is probably related to improving the balance between plasminogen activator and its inhibitory factor, increasing the activity of fibrinolysin, inactivating thrombin, inhibiting platelet aggregation induced by thrombin, and decreasing blood coagulation.     

穿心莲成分对血栓模型家兔血小板三磷酸肌醇及其受体表达的影响

目的　观察穿心莲成分 API0 1 34 (API)抗血小板活化和聚集的机制。方法　建立高脂血症家兔动脉血栓形成模型。观察穿心莲成分 API0 1 34 (API)对闭塞性血栓形成时间 (OT)、血小板聚集、血液血小板活化因子 (PAF)含量、血小板内三磷酸肌醇 (IP3)含量和 IP3受体 (IP3R)表达的影响。结果　 API能够显著延长 OT和抑制血小板聚集 ,降低血液 PAF和血小板内 IP3含量 ,抑制血小板内 IP3R蛋白的表达。API 5 0 mg/ kg的抑制作用明显强于 API 5 m g/ kg。结论　 API具有较强的抗血小板聚集和抗血栓形成作用 ,API对血小板 PAF- IP3/ IP3R信号途径的抑制作用 ,是 API抗血小板活化和聚集的机制之一     

API_(0134)对主动脉平滑肌细胞增殖的影响

采用体外培养的猪主动脉平滑肌细胞(SMC)为模型,在高脂血清(HLS)造成SMC增殖条件下,观察药物API_(0134)对SMC 增殖的作用及作用机制。结果发现:HLS 培养使 SMC 氚-胸腺嘧啶核苷(~3H-TdR)的掺入量增加;形态学上SMC 增生及合成功能增强。加用 API_(0134)后可以拮抗SMC的DNA 合成增强及形态学改变,同时降低脂质过氧化物(LPO),使前列环素(PGI_2),环磷酸腺苷(cAMP)含量增多,并使纤溶酶原激活物抑制物(PAI)活性降低。结果表明:API_(0134) 具有抑制SMC增殖的作用。     

超时间窗静脉溶栓对大鼠血栓性大脑中动脉栓塞的治疗效果及机制探讨

目的探讨超时间窗静脉溶栓对血栓性大脑中动脉栓塞大鼠的治疗效果。方法80只SD大鼠根据随机数字法分为对照组、脑梗死组、4.5 h溶栓组和6 h溶栓组,每组20只。对照组进行假手术处理,脑梗死组、4.5 h溶栓组和6 h溶栓组大鼠均建立大脑中动脉栓塞模型,4.5 h溶栓组和6 h溶栓组大鼠建模后分别于4.5 h、6 h给予重组组织型纤溶酶原激活剂（r-tPA）静脉泵入处理。测定各组大鼠脑梗死体积、微血管密度及皮层脑组织一氧化氮（NO）、丙二醛（MDA）含量、超氧化物歧化酶（SOD）活性。结果4.5 h溶栓组和6 h溶栓组3 d及7 d脑梗死体积均显著低于脑梗死组（P < 0.01）。4.5 h溶栓组和6 h溶栓组3 d及7 d脑微血管密度均显著高于脑梗死组（P < 0.01）,6 h溶栓组3 d及7 d脑微血管密度均低于4.5 h溶栓组（P < 0.05）。4.5 h和6 h溶栓组的7 d脑微血管密度均显著高于3 d脑微血管密度（P < 0.01）。与脑梗死组比较,对照组、4.5 h溶栓组、6 h溶栓组大鼠3 d和7 d皮层脑组织NO和MDA含量均明显降低,而SOD显著升高（P < 0.05~P < 0.01）,6 h溶栓组3 d和7 d皮层脑组织MDA含量高于4.5 h溶栓组（P < 0.05~P < 0.01）,6 h溶栓组3 d和7 d皮层脑组织SOD含量低于4.5 h溶栓组（P < 0.05）;4.5 h溶栓组和6 h溶栓组大鼠7 d皮层脑组织NO含量高于3 d NO含量（P < 0.05）。结论超时间窗静脉溶栓治疗可降低血栓性大脑中动脉栓塞大鼠的梗死体积,其机制可能与其可增加微血管密度及降低皮层脑组织NO含量和MDA含量、升高皮层脑组织SOD活性有关。     

The effects of trapidil and propranolol on platelet aggregation and prostacycline-thromboxane balance following acute myocardial infarction in rabbits

Summary Changes in infarct size, platelet aggregation, and plasma 6-k-PGF_{1 α} and thromboxane B₂ levels in the coronary sinus and arterial blood were observed in rabbits following acute myocardial infarction with or without trapidil or propranolol treatment. It was found that platelet aggregation, thromboxane B₂ and 6-k-PGF_{1 α} were elevated significantly in the control group 6 h after coronary ligation. These changes were inhibited by trapidil as well as propranolol, either of which could limit infarct size remarkably.     

丹酚酸B抗大鼠脑缺血作用及机制研究

目的观察丹酚酸B对缺血性脑卒中的保护作用及其机制。方法利用电凝法构建大脑中动脉栓塞模型,50只大鼠随机分为5组,空白对照组给予0.9%氯化钠注射液1 mL/kg;阳性对照组给予阿司匹林10 mg/kg;丹酚酸B低剂量组给予丹酚酸B 10 mg/kg;中剂量组给予丹酚酸B 20 mg/kg;高剂量组给予丹酚酸B 40 mg/kg,测量脑梗死面积、血栓重量,检测凝血酶原时间（PT）、活化部分凝血酶时间（APTT）、凝血酶时间（TT）、纤维蛋白原（FIB）。结果各组脑梗死面积及行为学评分差异均有统计学意义（P < 0.01和P < 0.05）。与空白对照组相比,不同剂量丹酚酸B脑梗死面积均明显缩小（P < 0.01）。5组血栓湿重及最大聚集率差异均有统计学意义（P < 0.01）。空白对照组血栓湿重均大于阳性对照组及丹酚酸B低、中、高剂量组（P < 0.01）,最大聚集率高于阳性对照组及丹酚酸B中、高剂量组（P < 0.01）。5组PT间差异无统计学意义（P>0.05）,TT、APTT、FIB差异有统计学意义（P < 0.05~P < 0.01）。结论丹酚酸B可通过抑制血小板聚集降低血栓形成,改善血液高凝状态,有效治疗缺血性脑卒中。     

高压氧综合治疗脑血栓形成50例临床观察

目的:观察50例脑血栓形成在常规治疗下加用高压氧治疗,对病人出现的临床症状和血液流变学的影响。方法:采用治疗组与对照组,常规使用甘露醇、复方丹参液、速避凝、抗菌素及营养脑神经细胞。治疗组:加用高压氧治疗。结果:治疗组明显优于对照组。结论:高压氧综合治疗能降低血液粘滞度,减少血小板的聚集,促进血栓的溶解吸收。     

导管溶栓联合支架植入术治疗急性髂股静脉血栓形成

目的比较经导管溶栓联合髂静脉支架植入与单纯导管溶栓治疗急性髂股静脉血栓形成的效果。方法选择经抗凝及导管溶栓治疗后存在严重髂静脉狭窄的急性髂股静脉血栓形成病人86例,其中32例经导管溶栓后,联合球囊扩张支架植入术开通髂静脉（观察组）;另54例单纯行导管溶栓治疗（对照组）。统计血栓消除率、肢体肿胀缓解情况及围手术期并发症;术后第3、6、12、24个月分别采用Villalta评分量表和Vcss量表评估血栓形成后综合征（PTS）发生情况及静脉临床症状严重程度。结果2组经导管溶栓治疗血栓Ⅲ级消除率达82.6%;溶栓时间、尿激酶用量、住院时间、围手术期并发症、PTS的发生率及血栓复发率差异均无统计学意义（P>0.05）;观察组膝上15 cm周径差较对照组小（P < 0.05）;随访3、6、12、24个月,观察组Villalta体征和症状评价、Vcss评分均低于对照组（P < 0.05~P < 0.01）。结论经导管溶栓治疗急性髂股静脉血栓形成血栓清除率高,联合球囊扩张支架植入术开通髂静脉有助于改善症状,提高临床治疗效果。     

Effect of Resveratrol on Preventing Steroid-induced Osteonecrosis in a Rabbit Model

Background:

Prevention of osteonecrosis (ON) has seldom been addressed. The purpose of this study was to evaluate the effect of resveratrol on preventing steroid-induced ON in rabbits.

Methods:

Seventy-two rabbits were divided into four groups: (1) NEC (ON) group: thirty rabbits were treated with lipopolysaccharide (LPS) once, then with methylprednisolone (MPS) daily for 3 days; (2) PRE (prevention) group: thirty rabbits were given one dose of LPS, then MPS daily for 3 days, and resveratrol on day 0 and daily for 2 weeks; (3) RES (resveratrol) group: six rabbits were given resveratrol for 2 weeks but without LPS/MPS; (4) CON (control) group: six rabbits were given alcohol for 2 weeks but without LPS/MPS. Levels of plasma tissue-type plasminogen activator (t-PA), plasminogen activator inhibitor 1 (PAI-1), thrombomodulin (TM), vascular endothelial growth factor (VEGF), maximum enhancement (ME) by magnetic resonance imaging, and ON incidence were evaluated.

Results:

The PRE group had a lower ON incidence than the NEC group, but with no significant differences at 2 weeks and 12 weeks. The RES and CON groups did not develop ON. TM and VEGF were significantly higher in the NEC group compared with the PRE group at weeks 1, 2, and 4 (TM: 1 week, P = 0.029; 2 weeks, P = 0.005; and 4 weeks, P = 0.047; VEGF: 1 week, P = 0.039; 2 weeks, P = 0.021; 4 weeks, P = 0.014), but the difference disappeared at 12 weeks. The levels of t-PA and PAI-1 were not significantly different between the NEC and PRE groups. The TM, t-PA, PAI-1, and VEGF concentrations in the RES and CON groups did not change over time. Compared to the baseline, ME in the NEC group decreased significantly (P = 0.025) at week 1, increased significantly (P = 0.021) at week 2, and was decreased at week 12. The variance was insignificant in the PRE group.

Conclusions:

Resveratrol may improve blood supply to bone in a rabbit model of ON of the femoral head via anti-inflammatory effects to protect the vascular endothelium and reduce thrombosis.     

人参总甙对正常家兔血液流变学的影响

本实验采用小剂量、长期给药的方法观察了人参总甙对正常家兔血液流变学的影响。结果表明人参总甙具有降低正常家兔血液粘度和血小板聚集性的作用,这可能是人参增加组织血流量、防治心血管疾患的重要依据。     

超声联合靶向微泡治疗急性脑血栓的动物实验研究

【摘要】 目的 探讨超声联合靶向微泡治疗急性血栓的效果,以及对纤溶系统的影响。方法 采用脑血管造影后将大白兔自体血栓通过导管注入颈动脉制备脑血栓动物模型。6 h后造影确定血栓未自溶。49只大白兔分为4组。A组（n=13）:经导管注射新型结合血小板Ⅱb /Ⅲa受体的靶向微泡（TMB）;B组（n=12）:经导管直接注射非靶向微泡（NTMB）;C组（n=12）:注射艾通立（重组纤溶酶原激活物r-TPA）:D组（n=12）:经导管注射生理盐水。A、B、D组均在治疗时应用低频超声(1 MHz,2.0 W/cm2)30 min。分别在治疗后立刻、1 h和2 h造影观察血栓溶解、血流再通情况,并在栓塞前和治疗后2 h取静脉血,检测凝血酶原时间（PT）、活化部分凝血酶时间（APTT）、凝血酶时间（TT）、纤维蛋白原（Fib）和D-二聚体。结果 A组6只（46.15%）,B组1只（8.33%）,C组4只（33.33%）,D组1只（8.33%）血栓溶解,A、C组血栓溶解率高于B、D组(P0.05),组间差异也无统计学意义(P>0.05),而C组中D-二聚体高于其他组（P<0.05）,Fib低于其他组（P<0.05）。结论 超声联合靶向微泡可快速溶解血栓使血管再通,其作用与 r-TPA相似而出血等副作用小。     

API0134对凝血酶作用下血小板环核苷酸和胞浆游离钙离子浓度的影响

为观察穿心莲有效成分（ＡＰＩ０１３４）对凝血酶作用下血小板活化的抑制作用,将家兔２４只随机分为ＡＰＩ０１３４（Ａ）组（ｎ＝９）,噻氯匹定（Ｔ）组（ｎ＝８）和对照组（ｎ＝７）,治疗１周,然后检测凝血酶作用下血小板内ｃＡＭＰ、ｃＧＭＰ和胞浆游离钙离子浓度,以及血小板超微结构。结果表明ｃＡＭＰ水平在Ａ、Ｔ两组显著高于Ｃ组（Ｐ＜０．０５）,而胞浆游离钙离子浓度明显低于Ｃ组（Ｐ＜０．０１）,超微结构表明Ａ组和Ｔ组血小板脱颗粒现象较Ｃ组明显减轻。在Ａ和Ｔ组之间各项参数均无统计学差异。结论：ＡＰＩ可抑制凝血酶诱导的血小板活化反应     

进展性脑梗死患者血浆相关血栓与止血参数变化的研究

目的对进展性脑梗死患者实验室凝血、血小板功能、抗凝和纤溶检测结果进行分析,为早期诊断和治疗进展性脑梗死提供参考依据。方法对117例进展性脑梗死患者进行前瞻性研究,检测血浆凝血酶原时间、凝血酶时间、部分凝血活酶时间、纤维蛋白原、血小板聚集率、抗凝血酶Ⅲ活性和组织型纤溶酶原激活物,并与同时期住院的666例完全性脑梗死患者进行对比。结果117例进展性脑梗死患者血浆凝血酶原时间、凝血酶时间和部分凝血活酶时间显著低于完全性脑梗死患者（P〈0.01）,纤维蛋白原明显高于完全性脑梗死患者（P〈0.05）,血小板聚集率显著高于完全性脑梗死患者（P〈0.01）,抗凝血酶Ⅲ活性和组织型纤溶酶原激活物水平显著低于完全性脑梗死患者（P〈0.01）。结论进展性脑梗死患者凝血功能较完全性脑梗死患者增强较为显著,其纤溶系统活动度明显降低,这些指标可作为临床上治疗进展性脑梗死的参考依据和预后的初步评估。     

脉血康胶囊对PCI术后急性冠脉综合症患者长期预后的改善作用

Objective:To study the changes of adenosine diphosphate(ADP)-induced platelet aggregation rate,and evaluate the effects of Maixuekang Capsule(脉血康胶囊,MKC) on platelet aggregation rate and long-term prognosis of patients with acute coronary syndrome after percutaneous coronary intervention(PCI).Methods:A total of 236 patients with acute coronary syndrome,who received successful PCI,were randomly assigned to a trial group(116 cases) and a control group(120 cases) according to random numbers;treatment allocation occurred when the participants met the inclusion criteria and signed the informed consent forms.In the trial group,the patients were treated with MKC combined with routine medication,and in the control group the patients were treated with routine medication.The therapeutic course for the two groups was 12 months and the follow-up was 12 months.The levels of ADP-induced platelet aggregation rate and serum high-sensitive C-reactive protein(hs-CRP) were determined before PCI,12 h and 30 days after PCI.In the meantime,the incidence of cardio-/cerebrovascular events was recorded during the 12-month follow-up.Results:Compared with before PCI,the levels of ADP-induced platelet aggregation rate and serum hs-CRP were significantly higher at 12 h after PCI(P0.05).They were significantly reduced after 30-day-treatment of MKC,showing statistical differences when compared with those in the control group(P0.05).During the 12-month follow-up,the incidence of cardio-/cerebrovascular events was significantly lower in the trial group than in the control group(6.9%vs.12.5%,P0.01).Conclusions:ADP-induced platelet aggregation function was significantly elevated after PCI.MKC improved the prognosis of patients with acute coronary syndrome,possibly through inhibiting the platelet aggregation,fighting against inflammation,and protecting the vascular endothelial function.     

API0134对血小板α颗粒膜蛋白和血小板聚集的影响

２０例急性心肌梗死患者在溶栓后随机分为穿心莲有效成分（ＡＰＩ０１３４,ＡＰＩ）组与对照组,治疗１５ｄ,均同时给予阿司匹林等药物。检测溶栓前后血浆α颗粒膜蛋白（ＧＭＰ－１４０）浓度和ＡＤＰ诱导的血小板聚集反应。结果表明,ＡＰＩ组溶栓后７２ｈ血浆ＧＭＰ－１４０浓度轻度增加,对照组显著性增加（Ｐ＜０．０１）。ＡＰＩ组ＧＭＰ－１４０在溶栓后７２ｈ和１５ｄ均低于对照组,差异有显著意义（均为Ｐ＜０．０５）。ＡＰＩ组１ｍｉｎ和５ｍｉｎ血小板聚集率均低于对照组,差异均有显著意义（７２ｈ,Ｐ＜０．０５;１５ｄ,Ｐ＜０．０１）。研究表明,ＡＰＩ与阿司匹林联合应用可抑制溶栓后血小板的活化,优于阿司匹林单独应用,因此ＡＰＩ有可能用于临床预防溶栓后的再闭塞。