Resveratrol inhibits periodontal pathogens in vitro

The gram‐negative anaerobic bacteria A. actinomycetemcomitans (Aa) and P. gingivalis (Pg) are key components in the aetiology of periodontal disease, and associated hard‐tissue destruction. Resveratrol is a phytoalexin, produced naturally by several plants when under attack by bacterial or fungal pathogens. It is found in many foods including mulberries, peanuts and the skin of labrusca and muscadine grapes. The objective of this study was to evaluate the effect of resveratrol on the in vitro growth of periodontal pathogens Aa and Pg. For comparison, resveratrol's effect on a variety of other oral microorganisms was also evaluated. Resveratrol demonstrates a poor solubility in water, thus different concentrations of resveratrol in the solvent dimethyl sulphoxide (DMSO) were added to calibrated suspensions of Aa and Pg. As a control, a parallel series of dilutions containing the vehicle DMSO alone was made to measure the effect of the solvent. Minimum inhibitory concentrations of the periodontal pathogens were calculated. All suspensions were incubated for 1, 3, 6 and 24 h in an anaerobic chamber at 37 °C. At each time interval, selected dilutions from each culture broth were plated on blood agar plates. Colonies appearing on blood agar plates were visually counted at 3 days for Aa, and at 5 days for Pg. The periodontal bacteria showed a significant decrease (p < 0.05) in viable counts after 1 h, whilst no colony forming units could be observed after 24 h. The results suggest that resveratrol possesses significant antimicrobial properties on periodontal pathogens in vitro. Copyright © 2011 John Wiley & Sons, Ltd.     

柚皮苷药理作用及机制的研究进展

柚皮苷是骨碎补、枳实、枳壳、化橘红等中药的主要药效成分,具有抗骨质疏松、抗氧化、抗炎、抗菌、抗肿瘤、改善心肌损伤、改善肝损伤、防治糖尿病及并发症和调血脂等药理作用。通过分析归纳有关柚皮苷的大量文献,对其药理作用及其机制的研究进展进行综述,旨在为其合理应用和药物开发提供参考。     

HPLC法测定活血胶囊中羟基红花黄色素A、柚皮苷

目的建立活血胶囊(黄芪、红花、桃仁、枳壳、川芎、当归、赤芍、酸枣仁等)中羟基红花黄色素A和柚皮苷测定方法。方法色谱柱Agilent HC-C18色谱柱(4.6 mm×150 mm,5μm);流动相甲醇-0.2%甲酸水(羟基红花黄色素A20∶80,柚皮苷33∶67),检测波长羟基红花黄色素A 403 nm,柚皮苷283 nm,体积流量0.8 mL/min,柱温25℃。结果羟基红花黄色素A在1.25～50μg/mL(R=0.999 9),柚皮苷在8.75～350μg/mL(R=1)质量浓度范围内峰面积具有良好的线性关系,方法回收率(n=6)分别为98.7%和99.2%,羟基红花黄色素A和柚皮苷的最低检出限分别为2.5 ng和2.75 ng。结论本方法操作简便,结果准确,重现性良好,可为活血胶囊的质量控制提供一定依据。     

不同产地枳壳中柚皮苷和新橙皮苷的测定

目的测定不同产地枳壳中柚皮苷和新橙皮苷的量,研究不同产地枳壳的质量差异。方法采用高效液相色谱法测定,色谱条件为Kromasil C18色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈-水(20∶80)(用磷酸调节pH值为3);体积流量1.0 mL/min;柱温30℃;检测波长283 nm。结果朱栾枳壳未检出柚皮苷和新橙皮苷成分;江枳壳、川枳壳、湘枳壳和常山胡柚均检出柚皮苷和新橙皮苷成分;陕西枳壳未检出新橙皮苷。江枳壳含7.5%柚皮苷、5.6%新橙皮皮苷,量均为最高。结论朱栾枳壳与其他产地枳壳的成分有差异,不含有《中国药典》2010年版规定的柚皮苷和新橙皮苷成分。     

柚皮苷通过Fas膜受体通路调控髓核细胞凋亡的机制研究

[目的]观察中药骨碎补单体柚皮苷(Naringin)对人退变椎间盘髓核细胞凋亡的影响,为治疗椎间盘退变提供理论依据。[方法]对体外培养的人退变椎间盘髓核细胞利用番红O染色和甲苯胺蓝染色进行鉴定,并对P3代细胞分别用柚皮苷0 g/m L(对照组)、柚皮苷20 g/m L、柚皮苷40 g/m L、柚皮苷80 g/m L的培养基培养人髓核细胞48 h,噻唑蓝(MTT)法选择其抑制人退变椎间盘髓核细胞凋亡的最佳浓度,并用流式细胞仪测定各组细胞凋亡率。实时定量聚合酶链锁反应(q RT-PCR)检测Fas、Fas L、Caspase-8基因表达。蛋白免疫印迹法(Western-blot)检测Fas蛋白表达。[结果]MTT法和流式细胞检测均测定20 g/m L柚皮苷为抑制髓核细胞凋亡的最佳浓度(P0.05)。q RT-PCR检测20 g/m L柚皮苷能够抑制Fas、Fas L、Caspase-8 m RNA表达(P0.05)。Western-blot检测显示20 g/m L柚皮苷能够抑制Fas、Fas L、Caspase-8蛋白表达。[结论]柚皮苷可能通过调控Fas/Fasl死亡受体凋亡通路抑制人退变椎间盘髓核细胞凋亡。     

Differential growth inhibitory effects of W. somnifera root and E. officinalis fruits on CHO cells

The Chinese Hamster ovary (CHO) cell line is widely used for measuring drug cytotoxicity and resistance. Therefore, the effects of two major Ayurvedic drugs (W. somnifera root and E. officinalis fruits) on the short and long-term growth of these cells were investigated. A standard 96-well plate assay was used to measure short-term growth. For assessment of long-term growth, the colony formation assay (CFA) was used, which measures clonogenic potential. This assay is the best measure of the cytotoxicity of anticancer drugs and the radio-sensitivity of tumor cells. As reported by others, the aqueous extracts of both herbal drugs were found to have short-term growth inhibitory effects on CHO cells when added to cells at the time of cell plating. However, this is the first report showing that these two herbal drugs have significantly different effects on the long-term growth of CHO cells. Thus, extracts of W. somnifera root, but not E. officinalis fruit, caused a reproducible, dose dependent, inhibition of colony formation of CHO cells.     

小蓟提取液对四种癌细胞生长抑制作用的研究

目的:研究小蓟提取液对癌细胞生长的抑制作用。方法:细胞形态观察和活细胞计数法研究小蓟提取液对人白血病细胞K562、肝癌细胞HepG2、宫颈癌细胞Hela、胃癌细胞BGC823生长的抑制作用。结果:小蓟提取液可使四种癌细胞发生皱缩、变圆、脱壁、碎裂等形态变化,生长受到明显抑制,抑制率最高可达88.27%。结论:小蓟有确切的抑癌作用,并与中医的气血理论、药物归经理论和肝肾同源理论相一致,再次说明了中医基本理论在体外细胞水平上也有良好体现,对实验室手段研究中医药同样有着重要指导意义。     

小檗碱抑制人成骨肉瘤细胞生长及机制研究

目的 探讨小檗碱对人成骨肉瘤细胞MG63生存率的影响.方法 将MG63细胞分为不加小檗碱的对照组和加小檗碱的实验组.采用CCK-8法测定各组细胞生存率,采用Western Blot法对小檗碱的作用机制进行探讨.结果 MG63细胞经0(对照组)、1、5、10、20、50 μg/ml浓度的小檗碱处理48 h后,细胞生存率分别为(99.37±1.13)％、(91.37±4.25)％、(72.44±2.89)％、(57.33±1.37)％、(50.14±4.05)％、(37.53±4.14)％.0(对照组)、20、50 μg/ml浓度的小檗碱处理MG63细胞48 h后,Bcl-2/β-action相对半定量表达值分别为(0.4167±0.0115),(0.3458±0.0146),(0.2987±0.0207);Bax/β-action相对半定量表达值分别为(0.4327±0.0206),(0.5007±0.0089),(0.7115±0.0214);Caspase-9/β-action相对半定量表达值分别为(0.1459±0.0107),(0.2416±0.0098),(0.2584±0.0330).实验组与对照组比较,差异均有统计学意义(P＜0.05).结论 小檗碱对MG63细胞具有抑制作用,其作用可能是通过调节凋亡基因Bcl-2、Bax的表达,活化caspases-9诱导肿瘤细胞凋亡.     

Observation on inhibitory effect of tetrandrine on the growth of human skin fibroblast in vitro]

The effect of tetrandrine on the growth of human skin fibroblasts was observed in vitro culture. The result revealed that tetrandrine significantly inhibited the growth of fibroblast. The higher the concentration of tetrandrine, the higher the inhibition rate, forming a time-dependent relationship. The possible value of tetrandrine in the treatment of hypertrophic scar is discussed.     

排脓散中桔梗对柚皮苷和芍药苷大鼠体内药动学的影响

目的比较排脓散中桔梗对大鼠体内柚皮苷和芍药苷的药动学影响,研究桔梗的配伍作用。方法 12只健康雄性SD大鼠随机分为排脓散全方组和排脓散缺桔梗方组,按16 g/kg ig给予排脓散全方提取物和排脓散缺桔梗方提取物,于不同时间点眼眶采血,以甘草苷为内标,采用超高效液相-质谱联用技术(UPLC-MS/MS)同时测定大鼠血浆中柚皮苷和芍药苷的浓度。以0.1%甲酸水溶液-甲醇为流动相梯度洗脱,体积流量0.2μL/min,采用电喷雾离子化(ESI)源,负离子方式扫描,多反应检测模式(MRM)检测2种指标成分,采用DAS2.0软件计算药动学参数。结果大鼠ig排脓散全方和排脓散缺桔梗方后,柚皮苷血液药动学参数曲线下面积(AUC0~t)分别为(2 643.22±277.75)、(1 934.01±371.98)μg·h/L,达峰浓度(Cmax)分别为(597.57±22.46)、(477.13±25.67)μg/L,清除率(CL)分别为(821.96±175.64)、(542.21±100.97)L/(h·kg),差异均具有显著性(P0.05);芍药苷血液药动学参数AUC0~t分别为(3 178.25±235.97)、(1 384.11±320.11)μg·h/L,Cmax分别为(816±28.88)、(402.67±58.22)μg/L,CL分别为(1 102.27±341.17)、(482.59±39.74)L/(h·kg),差异均具有显著性(P0.05)。结论配伍桔梗后,大鼠血浆中柚皮苷和芍药苷的药动学变化趋势大体相同,表现为达峰时间缩短,吸收程度和达峰浓度增加,清除速率加快,由此验证了桔梗在排脓散中的配伍具有科学合理性,同时为进一步研究排脓散的配伍规律提供参考。     

中药固金磨积片抗肝肺肿瘤移植和转移作用的实验研究

目的：探讨固金磨积片抗肿瘤转移作用。方法肝癌：应用小鼠移植肝癌22（腹水型）。按1：3比例制成瘤细胞悬液，每只小鼠0．2ml（约1～2×106）接种于NIH小鼠腹腔内，次日随机分为5组（n＝10），给药的3组分别按高（1．88g／kg），中（1．02g／kg）和低（0．592g／kg）三种剂量，每天1次灌胃给药，连续10d，停药后次日处死小鼠，抽腹水量，计算肿瘤细胞数。肺癌：取Lewis肺癌瘤组织悬液0．2ml接种于C57系小鼠右前腋皮下，24h后，随机分为5组（n＝10），用药组每天1次灌胃给药，剂量高、中、低三种（药量同肝癌组），连续20d，停药24h后处死小鼠，取肺组织在倒置显微镜下观察并计算肺转移瘤结节数。结果：①、固金磨积片对小鼠转性肝癌22瘤细胞生长有明显抑制作用，各用药组腹水形成缓慢，腹水量及每升腹水中所含的肿瘤细胞数与对照组比较均有明显差异（P＜0．05）。②、固金磨积片对Lewis肺癌转移结节有明显的抑制作用，与对照组比较（P＜0．05～0．01）。结论：本研究提示固金磨积片通过活血化瘀，清热解毒，扶正益气，保护和增强机体免疫功能，从而达到抑制肿瘤生长，抗肿瘤转移作用。     

藏药多刺绿绒蒿提取物体外抗流感病毒作用的初步研究

目的:研究藏药多刺绿绒蒿提取物体外抗流感病毒的作用;方法:制备了全草醇提取物;采用常规试管凝集实验方法,测试该提取物体外抗流感病毒活性;结果:发现该药液在1:4稀释度时(12.5mg/ml,按生药量计算),对体外甲型流感病毒有抑制作用。结论:多刺绿绒蒿提取液对流感病毒有一定的抗病毒活性,值得继续追踪研究。     

姜黄、生姜组方抑制幼龄大鼠前列腺生长的作用研究

目的观察姜黄、生姜组方乙醇提取物对正常幼龄大鼠前列腺生长的影响。方法将30只大鼠随机分为正常组(A组)、己烯雌酚组(B组)、姜黄生姜组(C组)、姜黄生姜蒲黄南瓜籽组(D组)、蒲黄南瓜籽组(E组),每组6只。除A组及B组外,C、D、E组分别饲以混有相应组乙醇提取物的饲料(10 g/d);B组每次按1.0 mg/kg体质量皮下注射己烯雌酚注射液,每周2次,连续3周,上述各组与A组均自由进食,连续3周。于末次给药24 h后处死动物,分离前列腺各叶、精囊腺和睾丸,分别称量,计算各腺体指数;并检测各组大鼠血清酸性磷酸酶水平。结果C组大鼠的前列腺背叶、侧叶、腹叶指数与A组相比减小,其中侧叶、背叶指数显著减小(P均<0.01),而睾丸指数区别不明显;D组前列腺背叶、侧叶、腹叶指数均减小,睾丸质量减轻。结论姜黄、生姜组方乙醇提取物能有效地降低大鼠前列腺的质量,对睾丸的生长无明显影响。     

In vitro inhibitory effects of bergenin and norbergenin on bovine adrenal tyrosine hydroxylase

The aqueous extract of Mallotus japonicus (Euphorbiaceae) showed an inhibitory effect on bovine adrenal tyrosine hydroxylase (TH), the rate-limiting enzyme in the biosynthesis of catecholamine. The present study was undertaken to investigate the effects of bergenin and norbergenin, constituents of the aqueous extract of Mallotus japonicus on bovine adrenal TH. Bergenin and norbergenin inhibited the TH activity by 29.0% and 53.4% at a concentration of 20 microg/mL, respectively, and exhibited noncompetitive inhibition of TH activity with the substrate l-tyrosine. The inhibition of TH activity and the inhibitory effect of norbergenin was more potent than that of bergenin. From these results, it is presumed that bergenin and norbergenin may be the active components of Mallotus japonicus in inhibiting TH, and these inhibitory effects may be partially responsible for the clinical use of Mallotus japonicus in treating peptic ulcer by reducing the availability of dopa/dopamine in vivo.     

3味中药对解脲支原体体外抑制作用的实验研究

目的　探讨中药野菊花、白花蛇舌草、车前草对解脲支原体 (Uu)的抑菌作用 ,为中药应用于临床治疗Uu感染 ,提供实验依据。方法　以水提法制备 3味中药原液 ,以琼脂稀释法测定野菊花、白花蛇舌草、车前草对 71株解脲支原体的最低抑菌浓度 (MIC)。结果　野菊花、白花蛇舌草对Uu具有明显的抑制作用 ,MIC均为 12 5mg/ml。14株标准菌株中未发现对此 2种中药的耐药菌株。车前草无明显的抑菌作用。三味中药均对细胞无明显毒性。结论　野菊花、白花蛇舌草在体外对Uu均有明显的抑制作用 ,车前草无明显抑菌作用     

Antitumor and immunoregulatory effects of astragalus on nasopharyngeal carcinoma in vivo and in vitro

This study was carried out to evaluate the effects of Astragalus on human nasopharyngeal carcinoma (NPC) viability and apoptosis and to investigate the mechanism of Astragalus in a NPC cell line (CNE2). Cell viability was measured using the MTT assay. CNE2 cells treated with Astragalus were stained with acridine orange/ethidium bromide and subjected to fluorescence microscopy. Bcl-2, Bax, caspase-3 and -8 were measured by western blotting. Rat NPC cells were used to establish a NPC model. Tumor weight, immune organ index and T lymphocyte subsets were employed to detect the immunoregulatory and antitumor effects of Astragalus after administration. Astragalus was effective in inducing apoptosis in CNE2 cells. Morphological changes associated with cell injury were found. Western analysis showed caspase-3, -8, and Bax protein levels were increased after Astragalus treatment, while the bcl-2 protein level was decreased. Astragalus increased the percentage of CD3(+) , CD4(+) T-lymphocytes, and the ratio of CD4(+) /CD8(+) . Astragalus also restored the immunological effects of DDP-induced immunosuppression. These findings suggest that the immunomodulatory and anticancer effects of DDP + Astragalus were better than those of DDP alone, and Astragalus could inhibit immunosuppression induced by DDP. The combination of CDDP + Astragalus could be developed as an effective chemotherapeutic regimen in the treatment of nasopharyngeal carcinoma.     

海力-5对家兔实验性动脉粥样硬化的抑制作用

海力－5系蒙医验方，具有暖胃消食、清糟归精；主治食欲不振，消化不良，胃脘冷痛，满闷嗳气。实验研究结果显示，对家兔血浆脂蛋白水平，超氧化物歧化酶含量和主动脉脂质斑块形成程度均有显著影响，能够显著抑制动脉粥样硬化。     

锦带花提取物的抑菌效果研究

锦带花为忍冬科植物锦带花Weigelaflorida(Bunge)A.DC.的干燥花蕾。具有活血止痛的功效,主治瘟病初起,头痛咽干、喉痹,痈肿疔疮,丹毒等。对锦带花提取物的抗菌作用进行初步研究,其结果表明:锦带花提取物对几种主要危害食品卫生、安全和对临床医学等具有重要影响的菌种,如金黄色葡萄球菌、绿脓杆菌、痢疾杆菌、乙型链球菌等均有一定抑制作用。     

青蒿素和青蒿琥酯对人乳腺癌MCF-7细胞的体外抑制作用比较研究

目的 对比研究青蒿素及其衍生物青蒿琥酯对人乳腺癌MCF—7细胞增殖的影响并探讨其作用机制。方法 采用体外培养的人乳腺癌MCF—7细胞株，利用SRB法测定青蒿素和青蒿琥酯对MCF—7细胞增殖的影响，FCM法测定细胞周期的变化，亚Gl期含量测定和DAN荧光染色法观察细胞凋亡。结果 10μmol/L青蒿素和lμmol/L青蒿琥酯能明显改变MCF—7细胞的细胞周期，使S期细胞显著减少，G0 Gl期细胞明显增加。青蒿素对MCF—7细胞增殖仅有微弱抑制作用，但其衍生物青蒿琥iB却有显著的抑制作用，IC50为0．3lμmol/L。同样，lμmol/L青蒿琥酷引起MCF—7细胞的凋亡和直接的细胞毒作用明显强于10μmol/L青蒿素的作用。结论 体外研究表明，对肿瘤细胞增殖的抑制青蒿琥酯比青蒿素作用强。     

3种骨碎补的品质评价研究

通过本草考证,商品调查,药材鉴定,弄清当前我国市售的骨碎补主流商品为水龙骨科植物槲蕨。建立了高效液相色谱法测定柚皮甙的含量。其平均回收率为99.5%,RSD为1.9%。