丹参对门脉高压血流动力学影响的实验与临床研究

为研究丹参对肝硬化门脉血流动力学的影响，利用血管插管测定用药前后胆管结扎肝硬化犬门脉系统压力变化，超声多普勒监测肝硬化患者用药后门静脉系统血流动力的改变。结果：（１）丹参静脉给药可使肝硬化犬门静脉压（ＰＰＶ）、嵌塞肝静脉压（ＷＨＶＰ）、肝静脉压力梯度（ＨＶＰＧ）显著降低（Ｐ＜０．０５～０．０１），而平均动脉压（ＭＡＰ）、心率（ＨＲ）无明显变化（Ｐ＞０．０５）；（２）丹参长期口服（１０～１２周），可显著降低肝硬化患者（Ｃｈｉｌｄ－ＰｕｇｈＡ、Ｂ级）门静脉内径（ＤＰＶ）、脾静脉内径（ＤＳＶ）、门静脉血流量（ＱＰＶ）、脾静脉血流量（ＱＳＶ）（Ｐ＜０．０５～０．０１），并对患者乏力、厌食、腹胀及肝功能（ＡＬＴ）具有部分改善作用，未见副作用。本研究表明，丹参为安全有效的降低门静脉压力药物，值得对其做进一步研究。     

丹参提取物改善大鼠重症急性胰腺炎肺损伤模型的ROS-NLRP3炎症小体信号通路机制

摘 要 目的：探讨活性氧簇（ROS） 核苷酸结合寡聚化结构域样受体蛋白3（NLRP3）炎症小体信号通路在丹参提取物改善急性胰腺炎肺损伤大鼠的作用机制。 方法: 60只SD大鼠随机分组5组：假手术组、模型组、丹参提取物（1.5 g·kg-1）组、ROS抑制剂组[N 乙酰 L 半胱氨酸（NAC）,20 mg·kg-1]、丹参提取物+NAC组（1.5 g·kg-1＋20 mg·kg-1）,每组12只。以逆行胆胰管注射5%牛磺胆酸钠（1 ml·kg-1）法建立急性胰腺炎肺损伤大鼠模型,各组灌胃给药,模型组和假手术组给予等量生理盐水,qd,持续5 d。处死大鼠,测量腹水量、右肺干质量、右肺湿质量,计算湿质量/干质量（W/D）比值,采用苏木精 伊红（HE）染色法对大鼠肺组织病理症状进行Holfbauer评分,以全自动血气分析检测仪检测动脉血中氧分压（PaO2）、二氧化碳分压（PaCO2）、氧合指数（OI）,用酶联免疫吸附（ELISA）法检测血清肿瘤坏死因子 α（TNF α）、白介素 6（IL 6）水平,ROS检测试剂盒检测肺组织中ROS水平,蛋白免疫印迹（Western blot）法检测NLRP3、半胱氨酸蛋白酶 1（caspase 1）、凋亡相关的斑点样蛋白（ASC）表达。 结果: 与假手术组比较,模型组大鼠腹水量、W/D比值、Holfbauer评分、PaCO2、TNF α、IL 6、ROS水平、NLRP3、caspase 1、ASC蛋白表达显著升高（P＜0.05）,PaO2、OI显著降低（P＜0.05）;与模型组比较,各药物处理组大鼠腹水量、W/D、Holfbauer评分、PaCO2、TNF α、IL 6、ROS水平、NLRP3、caspase 1、ASC蛋白表达显著降低（P＜0.05）,PaO2、OI显著升高（P＜0.05）;与丹参提取物组及NAC组分别比较,丹参提取物+NAC组大鼠腹水量、W/D、Holfbauer评分、PaCO2、TNF α、IL 6、ROS水平、NLRP3、caspase 1、ASC蛋白表达均显著降低（P＜0.05）,PaO2、OI显著升高（P＜0.05）。结论: 丹参提取物可以保护急性胰腺炎肺损伤大鼠的肺组织,可能通过下调ROS-NLRP3炎症小体信号实现。     

丹参诱导骨髓基质细胞神经元样细胞分化的microRNA-128路径

目的　研究microRNA-128在丹参制剂诱导大鼠骨髓基质细胞(bone marrow strowal cells,BMSCs)向神经元样细胞分化过程中的调控作用。  方法　培养鉴定大鼠BMSCs,应用反义寡居核苷酸转染技术（Lipofectamin 2000）转染microRNA-128 inhibitors于BMSCs;取转染和未转染的BMSCs,待细胞增殖到60%~70%融合时进行预诱导分化,预诱导24 h后,进行诱导分化;根据处理方式不同分为4组：未诱导组（A组）,丹参制剂诱导组（B组）,转染未诱导组（C组）,转染诱导组（D组）。流式细胞术鉴定BMSCs;qPCR检测microRNA-128及神经元特异性烯醇化酶（NSE）、巢蛋白（Nestin）、微管蛋白（β3-tubulin）的mRNA表达;Western blot检测NSE、Nestin和β3-tubulin的蛋白表达。  结果    BMSCs流式细胞检测CD29和CD90双阳性率达99.17%,CD11b和CD45双阴性率达99.21%;丹参制剂诱导及microRNA-128 inhibitors转染后,细胞microRNA-128表达量显著降低（P<0.05）;诱导与未诱导组,转染与未转染组比较,NSE、Nestin和β3-tubulin的mRNA和蛋白表达均显著升高（P<0.05）。  结论　丹参制剂能够抑制BMSCs的microRNA-128表达,进而影响BMSCs神经分化相关蛋白的表达,促进BMSCs向神经元样细胞分化。     

丹参多酚酸盐对持续性＋Gz 环境下大鼠牙髓的影响

目的：探讨丹参多酚酸盐（DSSM）对高正加速度（＋Gz）重复作用下的大鼠磨牙牙髓的作用。方法：27只 SD 雄性大鼠用随机区组法分为9组,每组3只。阳性对照组和药物组分别在＋5 Gz、＋10 Gz 的条件下共离心30 s,每天重复5次,间隔以＋1Gz 持续60 s,4 d /周,共3周。实验期间药物组于每次离心前30 min 按剂量腹腔注射 DSSM溶液（低、中、高给药剂量分别为2、6、18 mg/kg）,阴性及阳性对照组注射等体积生理盐水。实验结束后,光镜下观察各组大鼠牙髓组织形态学变化情况,并采用 Real time qPCR 方法测定大鼠磨牙牙髓中热休克蛋白（HSP70）mRNA 相对含量表达的情况。结果：＋5 Gz 和＋10 Gz阳性对照组大鼠活动程度降低,磨牙牙髓中出现空泡性变,组间差异不明显。HSP70 mRNA 的相对含量比阴性对照组高（P ＜0．05）。药物组中 HSP70 mRNA 的相对表达量较各自阳性对照组降低,＋10 Gz 高剂量组有统计学意义（P ＜0．05）。结论：＋Gz可引起牙髓损伤,＋Gz 值的增加与 HSP70 mRNA 相对表达量增加相关,DSSM可能对牙髓有保护作用。     

The interaction of ancrod with human platelets

Ancrod, a serine protease purified from the venom of Agkistrodon rhodostoma, has been used as a therapeutic anticoagulant for a number of indications, including replacement of heparin in patients with heparin-induced thrombocytopenia. Ancrod has similar fibrinolytic activity to thrombin, but ancrod specifically cleaves only the alpha chain of fibrinogen, producing the characteristic fibrinopeptides A, AP and AY. Because ancrod has been used in patients with heparin-induced thrombocytopenia, it is important to ensure that ancrod does not directly affect the platelets and potentially increase the hemostatic effect. The effect of ancrod on platelets has not been well established, and there is not agreement in published studies. Additionally, some of the studies are over 15 years old and pre-date sensitive assays such as glycoprotein analysis. For these reasons, we investigated the interaction of ancrod with human platelets using direct and indirect, functional and biochemical techniques. Incubation of platelets with ancrod alone did not induce platelet aggregation or the release of dense-granule contents. Pre-incubation of platelets with ancrod did not augment or inhibit the maximal aggregation achieved with thrombin, nor did it affect the amount of serotonin release from dense granules caused by activation by thrombin. Studies of ancrod-treated platelets using monoclonal antibody-mediated radioimmunoprecipitation demonstrated that high concentrations of ancrod did not cause measurable cleavage of either the glycoproteins Ib-IX or IIb-IIIa. Incubation of radiolabeled platelets with ancrod-treated plasma also had no effect on the platelet glycoproteins, indicating that ancrod does not indirectly affect the major surface receptors. Direct binding studies using radiolabeled ancrod did not demonstrate specific binding to the platelet surface. Together these studies indicate that ancrod does not directly affect nor bind to platelets in vitro . The hypocoagulant state and subsequent platelet function defect resulting from the use of ancrod appears to be limited to the removal of fibrinogen from the circulation.     

丹参对重度烧伤器官缺血-再灌注损伤保护作用的临床观察

[目的]探讨丹参对严重烧伤后重要脏器功能的治疗、促进创面愈合的临床疗效。[方法]选取2006年12月～2010年5月住院的74例重度烧伤病人,采用随机对照方法分为丹参治疗组(37例)和对照组(37例),治疗组应用注射用丹参冻干20mg/kg静滴,观察用药后5d、10d血常规、肝功能、肾功能和心肌酶谱、血清肿瘤坏死因子-α(TNF-α)及白介素-6(IL-6)水平,器官功能不全(OD)的发生率及病死率及平均愈合时间(d),并统计学分析。[结果]d5治疗组与对照组的各项检验指标改善有差异,P﹤0.05;d10部分检验指标改善有明显差异;治疗组5例出现OD,对照组10例死亡1例。[结论]注射用丹参冻干药用有活血化瘀的功效;烧伤早期大剂量使用可明显改善重要脏器的微循环,减轻缺血再灌注损伤,有效支持和保护重要脏器功能,能有效防止OD的发生、发展。     

Stephania tetrandra prevents and regresses liver fibrosis induced by carbon tetrachloride in rats

Background and Aim:  There is currently no safe and effective treatment for liver fibrosis. We have previously shown that Stephania tetrandra ( ST ) and Salvia miltiorrhiza ( SM ) suppress cell proliferation and enhance apoptosis of hepatic stellate cell (HSC) in vitro . In this study, we aimed to investigate the anti-fibrotic effect of these two herbs in vivo .
Methods:  Liver fibrosis was induced by carbon tetrachloride (CCl₄) injection in rats for 5 weeks. SM , ST or SM  +  ST was gavaged on day 1 of CCl₄ administration to study the preventive effects of herbs on hepatic fibrosis. In a separate study designed to assess possible fibrosis regression, rats were randomly allocated to be treated with SM, ST or SM  +  ST when fibrosis was established. Liver injury and collagen content were assessed. HSC activation and apoptosis were determined.
Results:  As compared with the CCL₄-only rats, serum ALT was significantly lower in CCl₄-treated rats that received either SM ( P  < 0.01) or ST ( P  < 0.01). Administration of ST significantly prevented ( P  < 0.01) or reversed the hepatic fibrosis ( P  < 0.01) induced by CCL₄. Moreover, rats treated with ST had reduced protein expression of α-SMA both in prevention ( P  < 0.05) and in regression ( P  < 0.01) experiments. The double-color staining of α-SMA and TUNEL showed that ST increased HSC apoptosis. However, co-treatment of SM  +  ST did not increase the antifibrotic effect of ST .
Conclusions:  Stephania tetrandra safely and effectively prevents and reverses hepatic fibrosis through activating HSC apoptosis in rats.     

二妙丸类方原方及经去多糖工艺处理后药效作用比较

目的比较二妙丸类方(二妙,三妙,四妙,全方)经去多糖工艺处理前后的药效作用。方法采用醋酸致小鼠扭体法,二甲苯致耳廓肿胀法,急性高尿酸法以及小鼠碳粒廓清法。结果经去多糖工艺处理以后,二妙丸类方的浓度得以提高,在用药剂量增加的情况下,使二妙丸类方的抗炎作用得以充分体现,其中二妙方的降尿酸作用增强,且二妙方组对非特异性免疫有一定的免疫增强作用。结论二妙丸类方的抗炎、镇痛、降尿酸等作用与所含多糖部分无关,可以在去多糖工艺处理的基础上,围绕抗炎、镇痛、降尿酸等药理作用继续开展二妙丸类方作用物质基础研究。     

正交试验优化白及粗多糖提取工艺

目的:优化白及粗多糖的提取工艺。方法:采用正交试验法,以白及粗多糖的提取率为指标,考察白及的粉碎粒度、提取时间和提取温度对提取工艺的影响。结果:最佳提取工艺为60目白及细粉分别以8、6和4倍量纯化水为溶剂提取3次,提取时间分别为3:2:2h,提取温度为90℃。结论:该提取工艺稳定可行,质量可控。