脉复生对兔动脉内皮细胞的作用研究

目的通过脉复生对兔动脉内皮细胞的作用研究,探讨脉复生对兔动脉内皮细胞的作用。方法在培养液中加入脉复生制剂的设为实验组,不加则为对照组,经培养后测定羟脯氨酸和糖胺多糖的含量。结果脉复生组中羟脯氨酸和糖胺多糖明显降低。结论脉复生有抑制动脉内皮细胞中羟脯氨酸和糖胺多糖的作用。     

山楂黄酮甙对牛内皮细胞作用的观察

本文以不同剂量的山楂黄酮M-199培养液培养牛内皮细胞,分别测定其细胞内、外液中羟脯胺酸和糖胺多糖的含量。结果表明:随着山楂黄酮甙剂量的增加,细胞内液和细胞外液中的羟脯氨酸含量均下降,与对照组比较,差异有非常显著意义(p<0.01);中剂量组的山楂黄酮甙促进糖胺多糖的代谢,而小剂量和大剂量组则抑制糖胺多糖的代谢,与对照组比较,差异均有非常显著性意义(p<0.01)。作者认为通过本试验,说明山楂黄酮甙在预防血栓形成和动脉粥样硬化的功能上有其药理学基础。     

黄芪多糖及丹酚酸对高糖诱导小鼠足细胞凋亡的影响

目的观察黄芪多糖与丹酚酸对高糖诱导的小鼠足细胞凋亡的影响。方法采用小鼠足细胞株,通过温度选择的培养方法诱导足细胞分化,分为正常对照组（5mmol/L）、甘露醇高渗对照组、高糖组、高糖＋黄芪多糖组、高糖＋丹酚酸组,分别于干预3h、6h、12h和24h后收集细胞;采用Annexin V-FITC和PI双染方法,用流式细胞仪观察各组足细胞的凋亡率。结果高糖组各时间点足细胞细胞凋亡率均高于同期正常对照组与高渗对照组（P〈0.01）;高糖＋黄芪多糖组、高糖＋丹酚酸组各时间点足细胞凋亡率均较高糖组降低（P〈0.05,P〈0.01）;高糖＋黄芪多糖组与高糖＋丹酚酸组比较差异无统计学意义（P〉0.05）。结论高糖可诱导足细胞凋亡,黄芪多糖与丹酚酸对高糖诱导的足细胞凋亡均有抑制作用。     

黄芪多糖对高糖刺激小鼠肾小球足细胞Desmin表达的影响

目的:体外模拟高糖病理微环境,探讨黄芪多糖对小鼠肾小球足细胞Desmin表达的影响。方法:将培养的小鼠肾小球足细胞分为:正常对照组、甘露醇高渗对照组、高糖组和黄芪多糖干预组,采用流式细胞术和real-Time PCR检测各组足细胞Desmin蛋白和mRNA的表达。结果:流式细胞术计数结果显示:黄芪多糖干预组小鼠肾小球足细胞表面Desmin阳性细胞百分率明显低于高糖组,差异有统计学意义(P0.05);较正常对照组和甘露醇高渗对照组,高糖组Desmin mRNA表达增加(P0.05);而相较高糖组,黄芪多糖干预组Desmin mRNA表达明显降低,差异有统计学意义(P0.05)。结论:在高糖刺激下,黄芪多糖干预可下调肾小球足细胞Desmin表达,是黄芪治疗DN蛋白尿的可能作用靶点。     

培养的鸡胚股骨中己糖醛酸和己糖胺的测定

已糖醛酸和已糖胺是软骨基质中糖胺多糖的主要成份,测定二者的含量以及它们的比例关系,对判定软骨的性质及糖胺多糖的性质是十分重要的。但是已糖醛酸在测化过程中容易遭受破坏,使测定值大大降低。用测定已糖胺的消化条件消化软骨,根本就不能用Bitter等改良的咔唑反应测定已糖醛     

复方附子多糖对肿瘤细胞糖基化作用的影响

目的观察中药多糖对糖蛋白糖链合成的不同糖基转移酶的诱导或阻遏作用,从而发现中药多糖的作用靶点及其作用机制。方法对含有附子多糖的不同药物分组,以肿瘤生长抑制率来观察复方多糖体内抗肿瘤作用,观察中药多糖对肝癌SK-HEP-1细胞糖基转移酶以及肿瘤相关基因表达的影响,同时借助多聚乳糖胺特异性生物素标记凝集素进行流式细胞术检测细胞多聚乳糖胺表达水平变化。结果与模型组比较,各给药组平均瘤重均显著降低(P0.01);与阿霉素组比较,3个复方组的平均瘤重无统计学意义(P0.05)。分别加附子多糖与中药复方多糖,多聚乳糖胺表达水平降低。结论多糖复方配伍抗肿瘤效果基本与阿霉素效果一致,且3个复方配伍组随着熟附子粗多糖比例的增加,降低了多聚乳糖胺表达水平,抗肿瘤作用更加明显。     

马齿苋多糖对高糖诱导的人晶状体上皮细胞发生上皮间质转化的影响

目的观察马齿苋多糖对高糖作用下永生系人晶状体上皮细胞(SRA01/04)上皮间质转化的影响。方法体外培养SRA01/04细胞,将传代的细胞随机分为正常对照组、高糖组、高糖+马齿苋多糖组,分别用含5.5mmol/L葡萄糖、30mmol/L葡萄糖、30mmol/L葡萄糖+0.5mg/mL马齿苋多糖的培养基培养48h,于培养的0h、6h、12h、24h、48h在倒置显微镜下分别观察三组的细胞形态学变化;48h后Western-blot检测细胞ZO-1、E-cadherin及α-SMA、Vimentin的蛋白表达变化。结果与对照组相比,高糖组细胞随时间延长细胞变长出现成纤维改变,细胞数量也逐渐减少;高糖联合马齿苋多糖组与正常组的形态变化基本趋于一致,没有随着时间延长而发生明显的梭形改变。与对照组相比,高糖组ZO-1、E-cadherin表达下降(P<0.01),α-SMA和Vimentin的表达上升(P<0.01);高糖联合马齿苋多糖组ZO-1、E-cadherin表达下降(P<0.05),α-SMA和Vimentin的表达上升(P<0.05)。与高糖组相比,高糖联合马齿苋多糖组ZO-1、E-cadherin表达上升(P<0.01),α-SMA和Vimentin的表达下降(P<0.01)。结论高糖可以诱导晶状体上皮细胞发生上皮间质转化,马齿苋多糖可以抑制这一过程。     

红花多糖对荷瘤鼠Ang-2、PTEN蛋白表达的影响

目的:研究红花多糖(SPS)对荷瘤鼠的移植瘤组织中Ang-2、PTEN蛋白表达的影响及意义。方法:选择昆明种小鼠80只制备荷瘤小鼠,随机分为4组,每组20只。对照组:模型组;实验组:环磷酰胺药物组、红花多糖+环磷酰胺药物联合组、受试药物红花多糖组。用药方法:(1)模型对照组给荷瘤鼠灌服生理盐水;(2)环磷酰胺药物组灌服环磷酰胺;(3)红花多糖+环磷酰胺药物联合组灌服环磷酰胺和红花多糖;(4)红花多糖组:红花多糖灌服荷瘤鼠。给药10天后停药,取肿瘤组织,称取湿重,计算抑瘤率。然后用S-P免疫组织化学法测定对照组、实验组荷瘤鼠移植瘤组织中Ang-2、PTEN蛋白的表达情况。结果:荷瘤鼠灌服生理盐水的模型对照组移植瘤组织中Ang-2的免疫组化阳性表达率显著高于各实验组(P0.05),而PTEN蛋白的阳性表达率显著低于各实验组(P0.01)。红花多糖、环磷酰胺联合用药组的抑瘤率最高,其次为环磷酰组、红花多糖组。结论:红花多糖具有抑制肿瘤组织生长的作用,可降低肿瘤组织中Ang-2的表达,从而增强PTEN蛋白的表达。     

黄芪多糖改善高脂高糖饮食大鼠认知障碍通过提高海马BDNF的表达

目的:研究黄芪多糖对高脂肪高糖饮食大鼠学习记忆能力的影响,探讨其可能的作用机制。方法:24只SD大鼠随机分为对照组、高脂高糖组和黄芪多糖组。高脂高糖组和黄芪多糖组饲以高脂高糖饲料,黄芪多糖组灌胃法给予黄芪多糖(60mg/kg)进行干预,Morris水迷宫方法检测大鼠学习记忆功能,电生理实验检测海马CA1区长时程增强,Westernblot方法检测海马BDNF的表达。结果:高脂高糖组水迷宫实验中探索潜伏期延长,记忆错误率增高,黄芪多糖干预能明显提高高脂高糖饮食大鼠水迷宫成绩及海马LTP,并且增加海马脑组织中BDNF的表达。结论:黄芪多糖能改善高脂高糖饮食导致的大鼠学习记忆能力和突触可塑性的下降,其机制与提高海马BDNF的表达有关。     

金钗石斛多糖对高糖诱导的大鼠肾系膜细胞氧化应激的影响

目的 观察金钗石斛多糖对高糖作用下大鼠肾小球系膜细胞(HBZY-1)氧化应激的影响.方法 采用水提醇沉法进行多糖的提取并经过DEAE-FF对提取粗多糖进行分离纯化;鉴定方法包括红外光谱分析该多糖的特征性吸收峰;紫外分光光度法对多糖纯度进行判定;苯酚-硫酸法进行多糖含量测定.后将HBZY-1细胞分为6组,对照组(NC)、高糖作用组(HG)、NAC组(NAC)、金钗石斛多糖低(LD)、中(MD)、高(HD)剂量干预组,药物作用时间24h,ABTS法检测各组细胞的总抗氧能力;流式细胞术检测细胞内活性氧的变化;WST-1法检测各组细胞的增殖情况.结果 经水提醇沉及阴离子交换柱纯化得到金钗石斛多糖组分FDP1,测得金钗石斛多糖相对含量为98.1％.与对照组相比,高糖作用组细胞的总抗氧能力下降,而ROS表达明显增加(P＜0.05).金钗石斛多糖干预组与高糖作用组相比,增加ROS生成.高糖作用组较正常组细胞的增殖速度加快(P＜0.05),金钗石斛多糖各剂量组对高糖诱导的细胞增殖无影响.结论 金钗石斛多糖FDP1具有抑制高糖诱导下的HBZY-1细胞总抗氧化能力的下降、ROS生成的增加,对高糖诱导下细胞氧化应激损伤有保护作用.     

六味地黄汤含药血清对兔软骨细胞表型的影响

目的:探讨六味地黄汤含药血清对关节软骨细胞增殖及表型的影响。方法:无菌条件分离新生24小时胎兔长骨干骺端透明软骨,用酶消化法分离原代关节软骨细胞,取P_2代软骨细胞分别种于96孔板(用于细胞增殖实验)和6孔板(用于细胞分泌性蛋白检测),设置空白对照组、正常兔血清组和六味地黄汤含药血清组,每组4个复孔。连续干预96小时后,CCK8法测定细胞增殖情况,ELISA法测定各组培养上清中Ⅱ型胶原(CollagenⅡ,colⅡ)、糖胺聚糖(glycosaminoglycan,GAG)含量。结果:干预结束后,CCK8法测定正常兔血清组和六味地黄汤含药血清组OD值均高于空白对照组,六味地黄汤含药血清组OD值高于正常兔血清组,差异均具有统计学意义(P0.05);ELISA法检测六味地黄汤含药血清组colⅡ、GAG含量均高于空白对照组,差异均具有统计学意义(P0.05);与正常兔血清组比较,六味地黄汤含药血清组GAG含量高于正常兔血清组,差异具有统计学意义(P0.05)。结论:六味地黄汤含药血清可以促进体外培养兔关节软骨细胞的增殖,促进软骨细胞分泌Ⅱ型胶原和GAG,从而更好地维持软骨细胞表型。     

Influence of Aloe vera on the glycosaminoglycans in the matrix of healing dermal wounds in rats

The influence of Aloe vera (L.) Burman f. on the glycosaminoglycan (GAG) components of the matrix in a healing wound was studied. Wound healing is a dynamic and complex sequence of events of which the major one is the synthesis of extracellular matrix components. The early stage of wound healing is characterized by the laying down of a provisional matrix, which is then followed by the formation of granulation tissue and synthesis of collagen and elastin. The provisional matrix or the ground substance consists of GAGs and proteoglycans (PGs), which are protein–GAG conjugates. In the present work, we have studied the influence of Aloe vera on the content of GAG and its types in the granulation tissue of healing wounds. We have also reported the levels of a few enzymes involved in matrix metabolism. The amount of ground substance synthesized was found to be higher in the treated wounds, and in particular, hyaluronic acid and dermatan sulphate levels were increased. The levels of the reported glycohydrolases were elevated on treatment with Aloe vera, indicating increased turnover of the matrix. Both topical and oral treatments with Aloe vera were found to have a positive influence on the synthesis of GAGs and thereby beneficially modulate wound healing.     

Yeast Hydrolysate Protects Cartilage via Stimulation of Type II Collagen Synthesis and Suppression of MMP‐13 Production

Type II collagen (COL II) is one of the primary components of hyaline cartilage and plays a key role in maintaining chondrocyte function. COL II is the principal target of destruction, and matrix metalloproteases (MMPs) have a major role in arthritis. In the present study, we investigated the chondroctye protection effects of specific fraction of yeast hydrolysate ((10–30 kDa molecular weight peptides). The mRNA expression of COL II was significantly increased in the YH‐treated group compared to the control at concentrations above 50 µg/ml, respectively. The 200 µg/ml YH‐treated group (3.43 ± 0.23 µg/ml) showed significantly reduced glycosaminoglycan (GAG) degradation relative to that in the interleukin‐1β (IL‐1β)‐treated control group (4.72 ± 0.05 µg/ml). In the YH‐treated group, MMP‐13 level was significantly decreased in a dose‐dependent manner compared to the IL‐1β‐treated group without YH treatment. However, MMP‐1 and MMP‐3 level were not different from that of control. Under the same conditions, we also examined mRNA levels of COL II. The mRNA expression of COL II was significantly higher in the YH‐treated group than in the IL‐1β‐treated control group at concentrations above 100 µg/ml. In conclusion, YH stimulated COL II synthesis and significantly inhibited MMP‐13 and GAG degradation caused by IL‐1β treatment. Copyright © 2012 John Wiley & Sons, Ltd.     

活血化淤药物764-3对胶原生物合成影响的研究

 目的：探讨活血化淤药物764-3对细胞外间质合成的影响。方法：羟脯氨酸法、间接酶联免疫吸附法（ELISA）测定博莱霉素A6（BLM-A6）诱发的小鼠、大鼠肺纤维化模型肺组织胶原含量及合成，阿利新蓝法测定其糖胺多糖（GAG）含量；³H-Pro掺入法测定体外培养兔角膜瘢痕成纤维细胞中胶原合成，ELISA测定其纤维粘连蛋白（Fn）含量。结果：764-3对模型肺组织胶原合成有明显的抑制作用，抑制率50．5％，尤其能使肺组织中Ⅰ型胶原及GAG明显减少；体外实验证实，764-3可抑制³H-Pro及³H-TdR掺入成纤维细胞，且使其表面Fn明显减少。结论：764-3抗纤维化作用机制的研究，应以细胞外间质组分——胶原合成为基点深入进行。     

Protective effects of berberine in an experimental rat osteoarthritis model

Berberine shows anticancer, antibacterial, antiinflammatory and antioxidant effects and may be useful in many clinical applications. The effects of berberine on articular cartilage metabolism remain unknown, so this study was performed to evaluate these effects in vitro and in vivo. For the in vitro work, rat articular chondrocytes were cultured in a monolayer and matrix metalloproteinase-1 (MMP-1), -3, -13 and tissue inhibitor of metalloproteinase (TIMP-1) expression was evaluated by real-time quantitative PCR. Nitric oxide (NO) levels were determined using the Griess reaction, and glycosaminoglycan (GAG) release was measured using the dimethylmethylene blue method. For the in vivo work, berberine was administered by intraarticular injection, and the effects on MMPs and TIMP-1 were examined at the gene and protein levels. Berberine was found to inhibit the expression of MMP-1, -3 and -13, and increased the level of TIMP-1 at the mRNA level in a dose-dependent manner. In IL-1β-induced rat articular chondrocytes, berberine decreased IL-1β-induced GAG release and NO production. Meanwhile, high-dose berberine exhibited an anticatabolic effect in an IL-1β-induced rat osteoarthritis (OA) model. These findings suggest that berberine may play a protective role in the development of OA and may be useful in the treatment of OA.     

Investigation of the Efficacy of Adjunctive Therapy with Bioavailability‐Boosted Curcuminoids in Major Depressive Disorder

Current medications have limited efficacy in controlling the symptoms of major depressive disorder (MDD), and are associated with several adverse events on long‐term use. Curcuminoids are extremely safe and multifunctional phytopharmaceuticals that have been shown to alleviate depressive symptoms in a variety of experimental models. The present study aimed to investigate the efficacy of curcuminoids as an add‐on to standard antidepressants in patients with MDD. One hundred and eleven subjects were assigned to standard antidepressive therapy plus curcuminoids–piperine combination (1000–10 mg/day; n = 61) or standard antidepressive therapy alone (n = 50) for a period of 6 weeks. Efficacy measures were changes in the psychological status on the basis of the Hospital Anxiety and Depression Scale (HADS) and Beck Depression Inventory II (BDI‐II). The BDI‐II and HADS total and subscale scores were reduced by the end of trial in both study groups. There were significantly greater reductions in total HADS score and subscales of anxiety and depression in the curcuminoids versus control group (p < 0.001). Likewise, reductions in BDI‐II total score and scores of somatic and cognitive subscales were found to be greater in the curcuminoids compared with control group (p < 0.001). Co‐administration of curcuminoids with piperine may be used as a safe and effective add‐on to standard antidepressants in patients with MDD. Copyright © 2014 John Wiley & Sons, Ltd.     

千斤拔属药用植物的研究进展

千斤拔入药始载于清朝吴其濬所编《植物名实图考》,是我国多民族使用、多基原药材。主要从千斤拔属药用植物的应用概况、化学成分、药理作用方面进行综述,为以后能够更加深入地研究千斤拔属药用植物提供依据,并且对其发展的市场潜力能有更直观的认识。迄今为止,千斤拔属植物中共有197个化合物被分离鉴定,结构类型主要包括黄酮类、皂苷及三萜类、蒽醌类和挥发油等。现代药理研究表明千斤拔属植物(主要为黄酮类)具有广泛的药理作用,包括镇痛与抗炎、抗菌、雌激素样、皮肤保护、神经保护、保肝等方面作用。千斤拔资源丰富,但其种质资源复杂,质量控制指标目前仍未统一,对千斤拔属植物进行更深入的多成分综合研究,将对进一步开发千斤拔属植物药用价值有重大意义。     

腰痛宁胶囊治疗腰肌纤维炎多中心临床试验研究

目的评价腰痛宁胶囊治疗腰肌纤维炎(寒湿瘀阻证)的临床有效性及安全性。方法采用随机、盲法、安慰剂平行对照、多中心临床试验方法,总病例数144例,其中治疗组72例,服腰痛宁胶囊;对照组72例,服用腰痛宁胶囊安慰剂。1个疗程即连续用药14 d后评价临床疗效及安全性。结果全分析集(FAS)分析显示,治疗组临床控制+显效例数之和与总例数的比值(控显率)优于对照组(P0.001),符合方案集(PPS)分析显示治疗组控显率优于对照组(P0.001);FAS分析显示,治疗组疼痛改善优于对照组(P0.001),PPS分析显示治疗组疼痛改善优于对照组(P0.001)。本试验共发生不良事件3例(2.10%),均发生在对照组,治疗组未发生不良事件,组间不良事件发生率无统计学差异(例数/例次P=0.245)。结论腰痛宁胶囊治疗腰肌纤维炎(寒湿瘀阻证)疗效确切,明显优于安慰剂,试验中未发现严重不良反应,安全性较好。