一种新型纤溶活性物质——纳豆激酶

<正>纳豆激酶（Natokinase）是一种能达到“经口纤溶疗法”目的、从世界数百种食物中筛选出来、安全性最高、在日本发现的高效血栓溶解酶。该酶是在把纳豆放在人工血栓上发现其作用的,目前其分子结构已经清楚。在血栓病国际会议上,有人可能不知道“纳豆”为何物,但却都知道纳豆激酶。     

极化液对冠心病患者纤溶系统功能的影响

目的：观察极化液葡萄糖－胰岛素－氯化钾（ＧＩＫ）用于冠心病治疗时对纤溶系统功能的影响。方法：４６例确诊冠心病患者,其中陈旧性心肌硬塞１０例,不稳定性心绞痛１４例,稳定性心绞痛２２例。给予ＧＩＫ液,即１０％葡萄糖５００ｍｌ＋普通胰岛素１０μｌ＋氯化钾１ｇ静滴 ８～１２ ｄ。于用药前后测定组织型纤溶酶原激活物活性（ｔ－ＰＡ：Ａ）及其抑制物活性（ＰＡＬ：Ａ）,纤溶酶原活性（ＰＬｇ）,组织型纤溶酶原激活物（ｔ－ＰＡ）及其抑制物（ＰＡＩ－Ｉ）含量及Ｄ－二聚体（Ｄ－Ｄｉｍｅｒ）含量,空腹血清胰岛素水平。结果：用药前与正常比较,ｔ－ＰＡ活性降低,ＰＡＩ－１活性增高,胰岛素水平增高,高胰岛素者占７８．２％。用药后ｔ－ＰＡ活性及含量、纤溶酶原活性均较前降低（Ｐ＜０．０１）,ＰＡＩ－１含量较前增加（Ｐ＜０．００１）,而ＰＡＩ－１活性及Ｄ－Ｄｉｍｅｒ变化不明显。结论：冠心病特别伴有胰岛素水平增高患者,应用ＧＩＫ液可能通过其对纤溶系统功能的不良影响促使病变进展。     

法莫替丁缓释片的研制及体外释药作用

研制了法莫替丁缓释片并测定其体外溶出度。结果表明：该制剂的体外溶出行为符合Ｈｉｇｕｃｈｉ方程，释药速度常数（Ｋ）主要受药物与辅料比例的影响；不同处方的释药速度常数（Ｋ）、释药５０％的时间、释药６３．２％的时间有显著差异（Ｐ＜０．０１）。     

豆豉中纳豆杆菌的筛选和纳豆激酶的初步分离

利用改良的培养基 纤维蛋白平板法 ,从我国传统食品豆豉中筛选出具有高纤溶活性的纳豆杆菌菌株 ;菌株液体发酵法制得粗酶液 ,采用沉淀法及柱层析等方法从发酵液中初步分离纳豆激酶 ;并应用体外溶栓实验考察分析了纳豆激酶的纤溶活性 ,为开发新型溶栓药物及保健食品提供了实验参考依据     

参三七皂甙Rg1对实验性血栓形成的影响及其机制探讨

用大鼠动静脉血栓形成模型，研究参三七皂甙Ｒｇ１抗血栓作用。结果表明，参三七皂甙Ｒｇ１可明显降低实验性血栓形成，对大鼠血浆纤溶系统亦有明显作用，可升高血浆中组织纤溶酶原激活物（ｔＰＡ）活性和活性型ｔＰＡ百分比，降低组织纤溶酶原激活物抑制剂（ＰＡＩ）活性。同时利用培养大鼠血管内皮细胞实验，发现Ｒｇ１可以剂量依赖性提高血管内皮细胞一氧化氮（ＮＯ）释放。提示Ｒｇ１抗血栓作用与增强纤溶系统活性，促进血管内皮ＮＯ释放有关     

甲壳胺-硝苯地平速释片的制备及生物利用度

报道了用硝苯地平（ＮＦ）和甲壳胺（ＣＴＳ）共同研磨后制备ＮＦ速释片的工艺．ＮＦ速释片与市售片剂的溶出度试验表明，其体外累积溶出百分率经０．５ｈ分别为８０％和不足２０％，人口服两片剂的对照试验表明，ＮＦ速释片组体内达峰明显缩短，生物利用度也有显著提高．     

界面活性剂对阿斯匹林片剂溶出速度的影响及其机制的研究

本文考察了三种不同类型界面活性剂月桂醇硫酸钠（ＳＤＳ），吐温－８０（ＴＥＮ）及十六烷基三甲基溴化铵（ＣＡＢ），对难溶性固体药物阿斯匹林（ＡＳＰＮ）片剂溶出速度的影响，并进一步探讨了其作用机制。实验表明界面活性剂ＣＡＢ和ＴＥＮ对ＡＳＰＮ具有改善润湿及反絮凝作用，可明显增加水中ＡＳＰＮ的溶解及其片剂的溶出，胶团增溶作用测定是很有限的，不能说明ＡＳＰＮ水中溶解的显著增加。界面活性剂ＳＤＳ对ＡＳＰＮ的润湿     

纳豆激酶的纤溶活性及其机理研究

目的研究纳豆激酶的抗血栓作用 ,探讨其主要作用机理。方法通过药理实验方法观察纳豆激酶对体内急性血栓形成 ,以及对血液凝固系统全血凝血时间、凝血酶原时间、凝血酶时间、活化部分凝血活酶时间和血液中纤维蛋白原含量变化的影响。结果纳豆激酶提取物可对抗小鼠体内血栓形成 ,对内外源性凝血途径均有影响 ,同时有直接的溶栓作用。结论纳豆激酶具有较强的纤溶活性。     

盐酸环丙沙星片的溶出度测定

盐酸环丙沙星片的溶出度测定ＩＮＶＥＳＴＩＧＡＴＩＯＮＯＮＤＩＳＳＯＬＵＴＩＯＮＯＦＣＩＰＲＯＦＬＯＸＡＣＩＮＨＹＤＲＯＣＬＯＲＩＤＥＴＡＢＬＥＴＳ童荣生＊陈素华张研林（四川省人民医院药剂科，成都６１００７２）ＴＯＮＧＲｏｎｇ－Ｓｈｅｎｇ＊，ＣＨＥＮＳ...     

醋酸泼尼松片的溶出度考察

醋酸泼尼松片的溶出度考察ＩＮＶＥＳＴＩＧＡＴＩＯＮＯＮＤＩＳＳＯＬＵＴＩＯＮＯＦＰＲＥＤＮＩＳＯＮＥＡＣＥＴＡＴＥＴＡＢＬＥＴＳ乔红王红雨燕朝丽（河南省南阳市药品检验所，南阳４７３０６１）ＱＩＡＯＨｏｎｇ，ＷＡＮＧＨｏｎｇ－Ｙｕ，ＹＡＮＣｈａｏ－Ｌｉ...     

来源于豆豉的纳豆激酶基因在大肠杆菌中活性表达研究

目的构建大肠杆菌表达质粒pTYB10 2 ,实现纳豆激酶基因 (nattokinasegene)在大肠杆菌中高活性表达。方法以纳豆芽孢杆菌基因组DNA为模板 ,采用PCR方法分别扩增编码前肽、成熟肽的序列 (pro NK) ,构建大肠杆菌表达质粒pTYB10 2 ,转化大肠杆菌ER2 5 6 6。在IPTG诱导下 ,分别在 37℃ (2h)、30℃ (3h)和 15℃ (14h)培养。结果pTYB10 2能表达有活性的纳豆激酶。SDS PAGE表明 ,15℃表达杂蛋白质更少。结论证明具有 77个氨基酸序列的前肽 (pro序列 )对纳豆激酶的活性表达是必不可少的。     

Effect of γ-aminobutyric acid and nattokinase-enriched fermented beans on the blood pressure of spontaneously hypertensive and normotensive Wistar–Kyoto rats

In this study we have evaluated the changes in arterial blood pressure in spontaneously hypertensive rats (SHR) caused by the short-term intake of Bacillus subtilis B060-fermented beans with significant γ-aminobutyric acid (GABA) and nattokinase activity. After being weaned, 7-week-old male SHR and 7-week-old male Wistar–Kyoto (WKY) rats were randomized into seven groups. Until the 8^th week of life, the rats in each group were given one of the following: Group 1, high dose of GABA and nattokinase in the SHR (SHD); Group 2, medium dose of GABA and nattokinase in the SHR (SMD); Group 3, low dose of GABA and nattokinase in the SHR (SLD); Group 4, negative control in the SHR (SD); Group 5, positive control in the SHR (SM); Group 6, high dose of GABA and nattokinase in the WKY (WHD); and Group 7, negative control in the WKY (WD). Distilled water served as the negative control, and captopril (50 mg/kg), a known ACE inhibitor, served as the positive control. Systolic blood pressure and diastolic blood pressure values were measured weekly from the 8^th week to the 16^th week of life using the tail-cuff method. A definite decrease in systolic and diastolic blood pressure values could be observed in the rats treated with captopril and in the rats that received GABA and nattokinase. The greatest antihypertensive effect was observed when the pharmacological treatment was administered. The effect of the daily intake of fermented beans containing GABA and nattokinase may be helpful in controlling blood pressure levels in hypertensive model animals. The fermentation of beans with B. subtilis B060 may therefore constitute a successful strategy for producing a functional food with antihypertensive activity.     

Effect of γ-aminobutyric acid and nattokinase-enriched fermented beans on the blood pressure of spontaneously hypertensive and normotensive Wistar–Kyoto rats

In this study we have evaluated the changes in arterial blood pressure in spontaneously hypertensive rats (SHR) caused by the short-term intake of Bacillus subtilis B060-fermented beans with significant γ-aminobutyric acid (GABA) and nattokinase activity. After being weaned, 7-week-old male SHR and 7-week-old male Wistar–Kyoto (WKY) rats were randomized into seven groups. Until the 8^th week of life, the rats in each group were given one of the following: Group 1, high dose of GABA and nattokinase in the SHR (SHD); Group 2, medium dose of GABA and nattokinase in the SHR (SMD); Group 3, low dose of GABA and nattokinase in the SHR (SLD); Group 4, negative control in the SHR (SD); Group 5, positive control in the SHR (SM); Group 6, high dose of GABA and nattokinase in the WKY (WHD); and Group 7, negative control in the WKY (WD). Distilled water served as the negative control, and captopril (50 mg/kg), a known ACE inhibitor, served as the positive control. Systolic blood pressure and diastolic blood pressure values were measured weekly from the 8^th week to the 16^th week of life using the tail-cuff method. A definite decrease in systolic and diastolic blood pressure values could be observed in the rats treated with captopril and in the rats that received GABA and nattokinase. The greatest antihypertensive effect was observed when the pharmacological treatment was administered. The effect of the daily intake of fermented beans containing GABA and nattokinase may be helpful in controlling blood pressure levels in hypertensive model animals. The fermentation of beans with B. subtilis B060 may therefore constitute a successful strategy for producing a functional food with antihypertensive activity.     

纳豆激酶固体浅盘发酵工艺的优化

目的以实验室优选的枯草杆菌为出发菌株,研究各种因素对菌株产酶的影响。方法通过单因素实验,确定固体浅盘发酵的最佳工艺条件。结果发酵工艺优化前酶活为12 0 4IU/g ,优化后可达到1931IU/g。结论固体浅盘发酵可获得高产量的纳豆激酶     

纳豆激酶的发酵工艺及其制剂学研究

目的研究纳豆激酶的发酵及其胶囊的制备工艺。方法考察纳豆激酶发酵提取液的热稳定性,并对纳豆激酶的制剂工艺进行研究。结果通过简单的制备工艺所制得的纳豆激酶胶囊,装量差异、含量均匀度、崩解时限均符合《中国药典》的要求。结论将纳豆激酶转化成可供临床应用的制剂具有重要意义。     

Antioxidation,angiotensin converting enzyme inhibition activity,nattokinase, and antihypertension of Bacillus subtilis (natto)-fermented pigeon pea

Because of the high incidence of cardiovascular diseases in Asian countries, traditional fermented foods from Asia have been increasingly investigated for antiatherosclerotic effects. This study investigated the production of nattokinase, a serine fibrinolytic enzyme, in pigeon pea by Bacillus subtilis fermentation. B. subtilis 14714, B. subtilis 14715, B. subtilis 14716, and B. subtilis 14718 were employed to produce nattokinase. The highest nattokinase activity in pigeon pea was obtained using B. subtilis 14715 fermentation for 32 hours. In addition, the levels of antioxidants (phenolics and flavonoids) and angiotensin converting enzyme inhibitory activity were increased in B. subtilis 14715-fermented pigeon pea, compared with those in nonfermented pigeon pea. In an animal model, we found that both water extracts of pigeon pea (100 mg/kg body weight) and water extracts of B. subtilis-fermented pigeon pea (100 mg/kg body weight) significantly improved systolic blood pressure (21 mmHg) and diastolic blood pressure (30 mmHg) in spontaneously hypertensive rats. These results suggest that Bacillus-fermented pigeon pea has benefits for cardiovascular health and can be developed as a new dietary supplement or functional food that prevents hypertension.     

纳豆激酶的分离纯化及酶学性质研究

目的研究纳豆激酶分离纯化工艺及酶学性质。方法纳豆激酶发酵液的粗提物经Superdex 75凝胶色谱和聚丙烯酰胺凝胶电泳(PAGE)分离纯化,采用TAME法测定酶的活性,通过SDS-PAGE对纯化结果进行了检验。结果SDS-PAGE中显示单一色带,相对分子质量28000,以TAME为底物时纳豆激酶的米氏常数(Km)为35.47mmol/L,最适宜的温度37℃,最适宜pH为8.6。结论该分离纯化方法可以得到较纯的纳豆激酶。     

纳豆激酶对实验动物溶栓作用的研究

目的观察纳豆激酶在体内溶解血栓的作用。方法利用30%FeCl3制造大鼠颈动脉血栓模型,随机分为5组,包括血栓对照组,低剂量纳豆激酶组,高剂量纳豆激酶组,蚓激酶对照组和生理盐水对照组。利用花生油喂食的方法构建高脂血症动物模型,分成4组进行试验,包括普通饲料对照组,添加花生油对照组,同时添加低剂量纳豆激酶和同时添加高剂量纳豆激酶组。结果血栓复制40 min后,纳豆激酶组与单纯血栓组相比,股动脉血压显著下降。此外,纳豆激酶组与单纯血栓组比较,经纳豆激酶处理的动物其血栓的湿质量、干质量降低。在高脂血症动物模型中,添加纳豆激酶组表现出总胆固醇(TC)和高密度脂蛋白胆固醇(HDL-C)明显降低,而且这种趋势随着纳豆激酶量的增高更加显著。结论纳豆激酶在体内具有明显的溶栓效果,并且有一定的预防和降低血脂的作用。     

纳豆激酶活性测定方法的研究

从专属性、灵敏度及实际操作的可行性等几个方面,对TAME法和纤维蛋白平板法进行了比较研究.结果：TAME法灵敏度高,操作简便,适合作为常规的分析检测手段,而平板法虽专属性高,却操作繁琐,适宜于定性分析.结论：利用TAME法与平板法各自的优点,将两者结合起来作为纳豆激酶的活性检测方法,可达到高效灵敏、专属性强的目的.