大蒜提取液与环丙沙星联合对铜绿假单胞菌PAO1与PAO-JP2的形态学影响

目的:采用银染法快速鉴定、对铜绿假单胞菌PAO1、PAO-JP2生物被膜形态学的比较,应用大蒜提取液和环丙沙星抗菌药,以研究QS系统影响铜绿假单胞形成生物被膜的机制。方法:分空白对照组,大蒜组(40ug/ml、80ug/ml),环丙沙星组(0.3ug/ml、0.5ug/ml、2ug/ml),两者混合组。结果:混合组联合对PAO1生物被膜抑制作用大于PAO-JP2。而且同一剂量下联合组对单独组对PAO1、PAO-JP2比较有显著差异(P0.05),结论:大蒜提取液对环丙沙星对PAO1、PAO-JP2生物被膜抑制起增强作用,从而抑制QS系统作用而减少耐药产生。     

大蒜提取液与环丙沙星杀菌活性的联合效应研究

目的:测定大蒜提取液与环丙沙星联合对铜绿假单胞菌PAO1、PAO-JP2生物被膜的影响进行研究,探讨联合用药对生物被膜细菌杀菌活性的作用。方法:采用棋盘法设计,分为空白对照组、大蒜提取液组、环丙沙星组、大蒜和环丙沙星组。结果:1/16、1/4MIC的大蒜提取液单独作用并不能使BF活菌数减少,但是1/16、1/4MIC的大蒜提取液却可以增强1/4MIC、1/2MIC、1MIC的环丙沙星对BF细菌的抑制作用。结论:大蒜提取液与环丙沙星联用对PAO1、PAO-JP2生物被膜抗菌有增效作用。     

大蒜提取液联合环丙沙星对铜绿假单胞菌PAO1、PAO-JP2浮游菌抗菌作用的研究

目的:测定大蒜提取液、喹诺酮类药物对PAO1、PAO-JP2浮游菌最低抑菌浓度,探讨两株菌对抗菌药物敏感性差异。方法:采用棋盘法设计,空白组、大蒜组、环丙沙星组、大蒜和环丙沙星组。结果:大蒜提取液对PAO1浮游菌MIC值高于PAO-JP2。结论:大蒜提取液和环丙沙星联用后,能明显降低该两种抗生素对铜绿假单胞菌PAO1、PAO-JP2的MIC值,大蒜提取液与环丙沙星联用后,抗菌效果以协同效应。     

大蒜提取液对铜绿假单胞菌LAS基因转录水平影响的研究

目的：研究大蒜提取液对铜绿假单胞菌LAS基因转录水平的影响。方法：通过在铜绿假单胞菌培养的阶段加入大蒜液,观察大蒜液对铜绿假单胞菌LAS基因转录水平的影响。结果：PAO-JP2组不能有效表达LasR基因,PAO-1组可有效表达LasR基因,但随着大蒜液亚抑菌浓度的增高,反义mRNA的表达量减少。结论：大蒜提取液可能是通过抑制LasR基因的表达的方式,抑制了QS系统。     

大蒜提取液的杀菌效果观察

目的:了解大蒜提取液对青霉菌,新型隐球菌和白假丝酵母菌的抑制作用。方法:使用K-B法对大蒜提取液对青霉菌、新型隐球菌和白假丝酵母菌的抑菌环直径进行测量。然后采用棋盘法分组对大蒜提取液联合环丙沙星对铜绿假单胞菌PAO1、PAO-JP2生物被膜细菌的影响进行实验测试,并进行比较分析。结果:大蒜提取液对青霉菌、新型隐球菌和白假丝酵母菌的抑制作用效果强弱依次为新型隐球菌、白假丝酵母菌和青霉菌。结论:大蒜提取液对青霉菌、新型隐球菌和白假丝酵母菌都有较强的抑制作用。其与环丙沙星联用,可以增强对铜绿假单胞菌PAO1、PAO-JP2生物被膜细菌的杀菌效果。     

穿心莲内酯对铜绿假单胞菌QS毒力因子的影响

目的:观察穿心莲对铜绿假单胞菌QS系统调控的毒力因子的影响。方法:分别测定12.5 mg.mL^-1穿心莲内酯对铜绿假单胞菌生长曲线、绿脓菌素、胞外蛋白水解酶和弹性蛋白酶活性的影响,同时检测穿心莲内酯对QS系统双突变株PAO-JP2株绿脓菌素、蛋白水解酶和弹性蛋白酶活性的作用。结果:穿心莲内酯对铜绿假单胞菌生长无明显影响。穿心莲内酯能明显抑制绿脓菌素的分泌、胞外蛋白水解酶和弹性蛋白酶活性。穿心莲内酯对PAO-JP2绿脓菌素、胞外蛋白水解酶和弹性蛋白酶活性无明显影响。结论:穿心莲内酯可能通过抑制铜绿假单胞菌毒力因子产生而实现抗感染作用。     

三七对铜绿假单胞菌密度感应相关毒力因子表达的影响

目的：通过观察三七对铜绿假单胞菌（Pseudomonas aeruginosa,PA）密度感应（Quorum sensing,QS）相关毒力因子表达变化的影响,探讨其作用机制。方法：用不同浓度三七与细菌共培养,采用弹性蛋白-刚果红降解试验测定弹性蛋白酶活性;采用紫外分光光度法测定绿脓菌素分泌量;结晶紫染色法观察生物被膜形成量;检测细菌在含三七琼脂平板中的集群运动与泳动能力。结果：三七在25mg/ml、12.5mg/ml和6.25mg/ml浓度下对弹性蛋白酶活性、绿脓菌素分泌有显著抑制作用,并能减少生物被膜形成。12.5mg/ml三七作用后细菌泳动能力受到抑制而集群运动无显著变化。结论：三七对铜绿假单胞菌密度感应相关毒力因子表达均有抑制作用,提示三七可以抑制铜绿假单胞菌密度感应功能,三七可能是铜绿假单胞菌密度感应抑制剂。     

桃红四物汤对蛋白酶激活受体激活诱导血小板释放VEGF和Endostatin的影响

目的考察桃红四物汤对蛋白酶激活受体(PARs)激动剂诱导人血小板聚集、释放血管内皮生长因子(VEGF)和内皮抑素(endostatin)以及释放液对内皮细胞增殖的作用。方法人富血小板血浆和不同质量浓度的桃红四物汤(0.5～2.5 mg/mL)孵育5 min后,加入PAR1选择性激动剂(SFLLRN-NH2,10μmol/L)或PAR4选择性激动剂(AYPGKF-NH2,200μmol/L)或凝血酶(1 U/mL),血小板聚集仪测定血小板聚集,ELISA测定血小板离心后上清中VEGF和endostatin,MTT测定释放液对内皮细胞增殖的作用。结果桃红四物汤在质量浓度0.5～2.5 mg/mL剂量依赖性的抑制凝血酶和PAR1-AP诱导的血小板聚集,但是对PAR4-AP诱导的血小板聚集没有显著的抑制作用;2.5 mg/mL桃红四物汤可抑制PAR1-AP诱导血小板释放VEGF以及PAR4-AP诱导endostatin释放;对PAR1-AP诱导释放液引起内皮细胞的增殖具有抑制作用,而对PAR4-AP诱导的释放液引起内皮细胞增殖减少具有促进作用。结论桃红四物汤抑制PAR1-AP诱导的血小板聚集,调节血小板释放VEGF和endostatin以及内皮细胞增殖可能是其发挥活血化瘀作用的机制之一。     

枇杷叶提取物对中性粒细胞弹性蛋白酶的抑制作用

目的探讨枇杷叶提取物体内外对人中性粒细胞弹性蛋白酶(HNE)的抑制作用。方法运用醇提方法获得枇杷叶提取物,测定其中总三萜成分的含有量。建立体外HNE抑制活性模型和体内烟雾致小鼠急性肺损伤模型,观察枇杷叶提取物通过HNE抑制通路对急性肺损伤的修复作用。结果枇杷叶提取物中总三萜含有量达到50.33%。体外实验中,枇杷叶提取物在质量浓度0.5~50.0μg/m L范围内,对HNE的抑制活性呈质量浓度依赖性,IC50为3.79μg/m L。在50、100、150 mg/kg 3个剂量下,枇杷叶提取能显著降低肺泡液TNF-α和小鼠中性粒细胞弹性蛋白酶(NE)水平。结论枇杷叶提取物可以通过抑制HNE的活性来降低急性肺损伤的程度。     

昆布多糖硫酸酯对非激素依赖型人前列腺癌细胞株PC-3相关基因蛋白表达的影响

目的研究昆布多糖硫酸酯(LAMS)对体外培养的非激素依赖型人前列腺癌细胞株PC-3相关基因蛋白表达的影响。方法利用Western blot检测人前列腺癌细胞增殖、凋亡相关基因PTEN、P27kip1、Caspase-3、BCL-2蛋白表达的结果。结果随着LAMS浓度的增加LAMS浓度的增加,BCL2蛋白的表达明显减弱,Caspase-3、p27和PTEN蛋白的表达明显增加,呈剂量依赖性。其中对照组(0ug/ml)与50μg/mL组间无显著性差异,100μg/mL组、200μg/mL组与对照组(0ug/ml)比较均有显著性差异(P<0.01)。结论 LAMS能影响PC-3细胞相关基因蛋白的表达,可能是其抑制前列腺癌的作用机制之一。     

盐酸小蘖碱抗幽门螺杆菌的实验及临床研究

目的:研究盐酸小蘖碱在体外的抗幽门螺杆菌(Helicobacter pylori,Hp)作用以及盐酸小蘖碱联合治疗Hp感染的临床疗效,为临床应用提供理论依据。方法:(1)胃镜下取25例快速尿素酶阳性患者的胃黏膜进行Hp临床株的分离纯化,连同12908、43504两例Hp标准株作为研究对象,将盐酸小蘖碱分为250μg、100μg、50μg、25μg四个浓度梯度组,以15μg标准克拉霉素纸片作为阳性对照,用纸片法进行体外药敏实验,比较平均抑菌圈直径及抗Hp有效率。(2)212例临床诊断为Hp感染的患者,随机分为4组:A组:PPI加克拉霉素加盐酸小蘖碱;B组:PPI加阿莫西林加盐酸小蘖碱;C组:PPI加阿莫西林加克拉霉素;D组:PPI加克拉霉素加甲硝唑。口服药物剂量:奥美拉唑20 mg·次~(-1)(其他PPI使用等效剂量),盐酸小蘖碱0.4 g·次~(-1),克拉霉素0.5 g·次~(-1),阿莫西林1.0 g·次~(-1),甲硝唑0.4 g·次~(-1),均2次·d~(-1)口服,连续服用7 d。抗Hp治疗停药4周后复查14 C呼气实验。比较Hp根除率、不良反应发生率。结果:(1)各浓度梯度组的盐酸小蘖碱抑菌圈直径均小于克拉霉素组,比较有统计学意义(P0.01)。盐酸小蘖碱250μg、100μg、50μg、25μg组与克拉霉素15μg组抗Hp有效率为92.59%(25/27)、85.19%(23/27)、44.44%(12/27)、44.44%(12/27)、88.89%(24/27),5组抗Hp有效率比较有统计学意义(P0.01),组间两两比较,克拉霉素15μg组与盐酸小蘖碱250μg、100μg组,盐酸小蘖碱250μg组与盐酸小蘖碱100μg组抗Hp有效率比较无统计学意义(P0.05)。(2)A组、B组、C组、D组Hp根除率分别为:70.37%(38/54)、61.22%(30/49)、68.63%(35/51)、65.52%(38/58),各组间根除率比较无统计学意义(P0.05);A组、B组、C组、D组不良反应发生率分别为9.12%、0.31%、2.87%、6.10%(P0.01);不良反应发生率组间比较:各组间差别均有统计学意义(P0.05)。结论:(1)盐酸小蘖碱在体外有良好的抗Hp作用;(2)临床应用PPI加盐酸小蘖碱加克拉霉素或阿莫西林组成三联方案的Hp根除率与对照组标准方案PPI加克拉霉素加甲硝唑或阿莫西林无明显差异;PPI加盐酸小蘖碱加阿莫西林组不良反应最少;含克拉霉素治疗组不良反应明显高于其他组。     

Antielastase and free radical scavenging activities of compounds from the stems of Cornus kousa

Bioassay‐guided investigation of the stems of Cornus kousa led to the isolation of 12 compounds such as four triterpenoids: betulinic acid (1), maslinic acid (2), arjunolic acid (3), 3‐isoarjunolic acid (4), four flavanoids; catechin (5), epi‐catechin (6), 2‐hydroxynaringenin (7), 2‐hydroxynaringenin‐7‐O‐β‐d ‐glucopyranoside (8), two ellagic acid derivatives; 3,4,3′‐tri‐O‐methylellagic acid (9), 3,4‐di‐O‐methylellagic acid (10), a daucosterol (11) and a sucrose derivative; (3′‐O‐p‐coumaroyl)‐β‐d ‐fructofuranosy‐(2?1)‐(6‐O‐p‐coumaroyl)‐α‐d ‐glucopyranoside (12). Their structures were elucidated on the basis of spectroscopic studies as well as by comparison with available data in the literature. The free radical scavenging activity and elastase inhibition activity were investigated for the development of antiaging ingredients as a raw material for use in cosmetics. Among these compounds, compounds 1, 2 and 8 showed significant elastase inhibition activity and IC₅₀ was 10.81 µg/mL, 21.21 µg/mL and 44.63 µg/mL, respectively, on porcine pancreatic elastase, whereas compounds 5, 6, 7 and 8 showed significant free radical scavenging activity and SC₅₀ was 8.37 µg/mL, 9.31 µg/mL, 9.23 µg/mL and 17.45 µg/mL, respectively. Copyright © 2007 John Wiley & Sons, Ltd.     

大蒜3个有效部位配伍对3种人胃癌细胞的杀伤作用

目的:观察大蒜3个有效部位组合物对3种人胃癌细胞的杀伤作用,筛选有效部位组合物最佳配伍比例.方法:以大蒜3个有效部位(大蒜油、大蒜总多糖、大蒜总皂苷)为研究对象,选用L16(45)正交表设计分组、给药,以胃癌细胞代谢MTT活力(抑瘤百分率)作为考察指标,采用SPSS软件对实验数据进行处理.结果:大蒜3个有效部位对MKN45细胞杀伤作用的主次顺序为大蒜油(1#)＞大蒜总多糖(2#)＞大蒜总皂苷(5#),其最佳配比为1 #∶2#∶5#=1∶20∶10.大蒜3个有效部位对AGS细胞抑制率影响无明显差别,其最佳配比为1#∶2#∶5#=1∶13.3∶3.3.大蒜总皂苷对人胃癌HGC-27细胞抑制率的影响显著(P＜0.01),影响的主次顺序为大蒜总皂苷＞大蒜油＞大蒜总多糖,其最佳配比为1#∶2#∶5#=1∶200∶50.结论:3种人胃癌细胞对大蒜3个有效部位组合物的杀伤作用的敏感性不同,3个有效部位组合对3种人胃癌细胞杀伤作用的最佳配比浓度亦不相同.     

Hypochlorous acid scavenging capacity of garlic

The ability of the following aqueous garlic extracts to scavenge hypochlorous acid (HOCl) was studied: (a) unheated extract of raw garlic (RG), (b) heated extract of raw garlic (HRG), (c) extract of boiled garlic cloves (BG) and (d) extract of microwave-treated garlic cloves (MG). The HOCl scavenging activity was evaluated following changes in the absorption spectrum of catalase (370-450 nm). RG, HRG and BG scavenged HOCl in a concentration-dependent way: the IC(50) (mg/mL) values for RG, HRG and BG were 0.339 +/- 0.028, 0.632 +/- 0.030 and 0.690 +/- 0.031, respectively. The HOCl scavenging ability of garlic was significantly decreased when garlic cloves or the extract of raw garlic were heated (BG and HRG, respectively); this scavenging ability was eliminated when garlic was subjected to microwave heating (MG). The IC(50) value for the reference compound, ascorbic acid, was 0.500 +/- 0.049 mg/mL. In summary, raw garlic concentration-dependently scavenged HOCl; this ability was eliminated in extracts of microwave-treated garlic cloves but not in heated extracts of raw garlic or extracts of boiled garlic cloves.     

The effect of garlic consumption on Th1/Th2 cytokines in phytohemagglutinin (PHA) activated rat spleen lymphocytes

The balance and regulation of T helper 1 (Th1) and Th2‐type cytokines are important in the effective immune response to different diseases. To clarify the effect of garlic (Allium sativum L.) consumption on the Th1/Th2 balance, the secretion of gamma interferon (IFN‐γ) and interleukin‐4 (IL‐4), as two prototypes of Th1/Th2 cytokines, were compared in serum and supernatant of in vitro phytohemagglutinin activated rat spleen lymphocytes. Thirty male rats were divided equally into two groups. The treatment group received garlic solution in water (600 mg/kg/4 mL) and controls received distilled water by gavage. After 1 month, serum and supernatant of PHA activated spleen lymphocytes were analysed for IFN‐γ and IL‐4 by the enzyme‐linked immunosorbent assay test and thymus and spleen weights were measured. The garlic treatment group showed significantly decreased production of IFN‐γ from 101.73 ± 4.62 to 74.64 ± 4.64 pg/mL and significantly increased IL‐4 production from 26.75 ± 3.35 to 83.92 ± 6.56 pg/mL (p < 0.001) in the supernatant of PHA induced spleen lymphocytes. The serum level of these cytokines was undetectable. The mean weight of thymuses in the garlic fed animals was significantly reduced from 0.456 ± 0.016 to 0.368 ± 0.023g compared with the control group (p < 0.005). There were no significant differences between the spleen weights in the two groups. In conclusion, oral garlic treatment may favor a Th2 or humoral immune response. Copyright © 2008 John Wiley & Sons, Ltd.     

虎杖大黄素与黄芪总多糖合用抗Ⅱ型人疱疹病毒药效分析

目的 :为了探讨中药组方时有效成分间的相互作用对药效的影响 ,我们进行了此项研究。方法 :我们以阿昔洛韦 (ACV)为阳性对照 ,采用空斑减数实验 ,用中效作用原理对虎杖大黄素与黄芪总多糖合用抗HSV 2 333株药效进行了分析。结果 :在Hep 2细胞系统中 ,ACV对HSV 2 333株增殖抑制的ED5 0为 10 85ug ml。当大黄素与总多糖以 1(Dm)大黄素 1(Dm)总多糖的比例合用时 ,对HSV 2 333株直接杀灭、感染阻断、增殖抑制的ED5 0依次为 1 5 5ug ml ,0 85ug ml,1.18ug ml;合用指数 (CombinationIndex ,CI)均 <1;相关系数均为 0 99,P <0 .0 0 0 5 ;说明这一合用比对HSV 2333株具有明显的协同对抗作用。结论 :ACV对HSV 2 333株增殖抑制的治疗指数 (TreatmentIndex ,TI)为 10 0。 1(Dm)大黄素 1(Dm)总多糖对HSV 2 333株直接杀灭、感染阻断、增殖抑制的TIs为340 9,6 2 1 6 ,4 47 8,是最佳合用比。     

Compounds with tyrosinase inhibition, elastase inhibition and DPPH radical scavenging activities from the branches of Distylium racemosum Sieb. et Zucc

Twenty compounds were isolated from the ethanol extract of Distylium racemosum branches and their inhibitory activities on tyrosinase, elastase and free radicals evaluated. The isolated compounds were identified as dibenzofurans (1–4), abscisic acid (5), 6′‐O‐galloylsalidroside (6), catechin derivatives (7–11), gallic acid derivatives (12–14), tyrosol (15), flavonoids (16–18), lupeol (19) and 1,2,3,6‐tetragalloylglucose (20). For study of tyrosinase inhibition activities, when compared with arbutin (IC₅₀ 48.8 μg/mL), four compounds (8, 11, 13, 17) showed higher activities, with IC₅₀ values of 4.8, 30.2, 40.5 and 37.7 μg/mL, respectively. For the elastase inhibition test, dibenzofuran 1 showed greater activity than the positive control, oleanolic acid (IC₅₀ 9.7 μg/mL), with an IC₅₀ of 7.7 μg/mL. In the studies on DPPH radical scavenging activities, five compounds (11, 12, 13, 14, 15) showed higher activities than ascorbic acid (IC₅₀ 5.0 μg/mL), with IC₅₀ values of 4.6, 3.9, 2.9, 3.8 and 4.7 μg/mL, respectively. Copyright © 2011 John Wiley & Sons, Ltd.