咖啡酸锗对小鼠U14瘤的抑制作用及其免疫功能的实验研究

目的探讨咖啡酸锗对小鼠U14瘤的抑制作用及其对免疫功能的影响。方法采用小鼠U14瘤模型,以咖啡酸锗尾静脉注射给药,观察抑瘤率,并测定小鼠胸腺指数和脾脏指数及外周血淋巴细胞酸性a-醋酸奈酯酶(ANAE)的阳性率。结果咖啡酸锗对小鼠U14瘤具有明显抑制作用,高、中、低剂量组抑瘤率分别为45.47%、57.86%、55.83%(P<0.01);各剂量组胸腺指数和脾脏指数与生理盐水组比较无明显差异,与环磷酰胺组比较有显著性差异(P<0.01)。ANAE染色标记T淋巴细胞,环磷酰胺组显著低于正常组及咖啡酸锗各剂量组(P<0.01);咖啡酸锗各剂量组较生理盐水组比较无明显差异。结论咖啡酸锗对小鼠U14瘤有明显的抑制作用而对免疫功能无明显影响。     

咖啡酸锗对小鼠U14瘤的抑制作用及其免疫功能的实验研究

目的 探讨咖啡酸锗对小鼠U14瘤的抑制作用及其对免疫功能的影响.方法 采用小鼠U14瘤模型,以咖啡酸锗尾静脉注射给药,观察抑瘤率,并测定小鼠胸腺指数和脾脏指数及外周血淋巴细胞酸性a-醋酸奈酯酶(ANAE)的阳性率.结果 咖啡酸锗对小鼠U14瘤具有明显抑制作用,高、中、低剂量组抑瘤率分别为45.47%、57.86%、55.83%(P&lt;0.01);各剂量组胸腺指数和脾脏指数与生理盐水组比较无明显差异,与环磷酰胺组比较有显著性差异(P&lt;0.01).ANAE染色标记T淋巴细胞,环磷酰胺组显著低于正常组及咖啡酸锗各剂量组(P&lt;0.01);咖啡酸锗各剂量组较生理盐水组比较无明显差异.结论 咖啡酸锗对小鼠U14瘤有明显的抑制作用而对免疫功能无明显影响.     

咖啡酸锗对小鼠肝癌H22抑制作用的研究

目的:探讨一种新的有机锗化合物(咖啡酸锗)对小鼠肝癌H22的抑制作用。方法:通过体内抗肿瘤实验,观察咖啡酸锗对小鼠肝癌H22的影响(抑瘤率),并采用HE染色和迈-格-姬(MGG)染色在光镜下观察肿瘤细胞的病理形态变化。结果:咖啡酸锗能有效地抑制小鼠肝癌H22的生长,咖啡酸锗低、中、高剂量组的抑瘤率分别为46.48%、50.00%、52.11%,光镜下见咖啡酸锗各剂量组肿瘤细胞生长受抑,浸润程度、核分裂数、血管数目明显减少,而且坏死程度较严重。结论:咖啡酸锗对小鼠肝癌H22具有显著的抑制作用。     

原位末端标记技术检测咖啡酸锗对小鼠宫颈癌U14瘤的凋亡诱导作用

目的应用脱氧核苷酸转移酶介导的原位末端标记技术(TUNEL),检测咖啡酸锗对小鼠宫颈癌U14瘤的凋亡诱导作用。方法建立小鼠宫颈癌U14移植瘤动物模型,观察抑瘤率,并应用TUNEL检测各组瘤细胞凋亡阳性率。结果咖啡酸锗对小鼠有明显抑制作用,其高、中、低剂量分别为45.47%、57.86%、55.83%,差异有统计学意义(P0.01),TUNEL显示咖啡酸锗低剂量组细胞凋亡阳性率最高,达80.00%,与对照组比较差异有统计学意义(P0.01)。结论咖啡酸锗对小鼠U14瘤有明显的抑制作用,可诱导U14移植瘤细胞凋亡,可能是其抑癌机制之一。     

加味小陷胸汤抗肿瘤作用的实验研究

目的:研究加昧小陷胸汤的抗肿瘤作用.方法:用ICR小鼠复制S180实体瘤和ESC腹水瘤模型,采用经口灌胃给药法,分别观察其对小鼠移植性肿瘤生长与荷瘤生存时间的影响;用炭粒廓清法检测其对荷瘤小鼠单核-巨噬细胞系(MPS)吞噬功能的影响.结果:加味小陷胸汤中、高剂量对S180小鼠肉瘤生长的抑制率分别为34.708%和50.31%;对荷ESC腹水瘤小鼠的生命延长率分别为43.47%和53.26%(P＜0.05);能明显促进荷瘤小鼠MPS吞噬功能(P＜0.05或＜0.01).结论:加味小陷胸汤对小鼠移植性肿瘤S180有一定抑制作用,能明显延长荷瘤小鼠存活时间,并能明显促进荷瘤小鼠非特异性细胞免疫功能.     

异臭椿烷对S180肉瘤细胞小鼠移植瘤的影响

目的探讨不同剂量的异臭椿烷治疗S180肉瘤细胞小鼠皮下移植瘤效果。方法建立S180肉瘤细胞小鼠皮下移植瘤动物模型,随机分为空白对照组、阳性对照组、异臭椿烷高、中、低剂量组,每组9只。异臭椿烷高、中、低剂量组分别给予1.0、0.5、0.1mg/L异臭椿烷0.1mL,灌胃给药,每天1次;阳性对照组采用环磷酰胺腹腔注射给药,25mg/（kg·d）;空白对照组每只给予0.1mL纯食用油灌胃。5组均连续给药5d后间隔2d再次连续给药5d。停药后24h处死小鼠,称取小鼠体质量,比较小鼠实验前后体质量变化;称取瘤质量,计算抑瘤率。结果异臭椿烷各剂量组和阳性对照组小鼠状态总体表现明显强于空白对照组,体质量增加幅度明显高于空白对照组,肿瘤生长速度明显慢于空白对照组,差异均有显著意义（F=34.786、16.407,q=3.059～16.077,P〈0.05）。结论异臭椿烷对S180肉瘤细胞小鼠皮下移植瘤生长具有一定的抑制作用。     

咖啡酸锗诱导小鼠宫颈癌(U14)细胞凋亡的实验研究

目的:从分子水平探讨咖啡酸锗对小鼠宫颈癌(U14)细胞的抑制效应及作用机制.方法:抗肿瘤活性实验采用:(1)药理试验;(2)普通染色光镜观察;(3)免疫组化观察PCNA、p53、bcl-2、ER、PR的表达;(4)甲基绿.派诺宁染色法;(5)流式细胞仪(FCM)测定DNA含量.结果:(1)咖啡酸锗及环磷酰胺组瘤重明显减轻,抑瘤率分别为47.28%和54.27%;(2)咖啡酸锗及环磷酰胺组的凋亡率分别为23.32%和20.61%;(3)咖啡酸锗组在DNA直方图上出现"亚G1"峰(即凋亡峰),生理盐水组无明显"亚G1"峰;咖啡酸锗对U14细胞周期有明显影响,使U14细胞G2～M期减少、细胞周期延长、对细胞增殖的抑制作用增强.结论:咖啡酸锗对小鼠宫颈癌(U14)细胞生长有一定的抑制作用,诱导瘤细胞凋亡可能是其抑制U14细胞生长的机制之一.     

半枝莲水提取物调节肿瘤VEGF/DC实验研究

目的研究半枝莲水提取物体内抗肿瘤及其对VEGF表达和DC浸润的调节作用。方法动物移植性肿瘤实验观察半枝莲水提取物、半枝莲水煎剂对小对鼠体内肿瘤细胞生长和胸腺、脾、肝脏的影响,采用免疫组化技术检测半枝莲水提取物对瘤体内VEGF表达及DC浸润的影响。结果半枝莲水提取物对S180肉瘤生长的抑制率以中剂量为佳;高、中剂量组脾脏指数、肝脏指数与模型组相比较均有显著性差异。各剂量组胸腺指数与模型组相比均有显著性差异,高剂量(200 mg/kg)对胸腺有抑制作用,各用药组能够降低S180小鼠肿瘤组织中VEGF的表达,提高DC的浸润,以中剂量最为显著。结论半枝莲水提取物在体内具有抑制S180肉瘤的作用,并能增强荷瘤小鼠的免疫能力,VEGF的表达与DC浸润成负相关,有下调VEGF和上调DC的作用。     

生半夏水提取液对小鼠中枢神经抑制作用的研究

目的 研究生半夏水提取液给小鼠灌胃后对中枢神经的抑制作用.方法 将小鼠随机分为3组,即空白对照组,实验组和阳性对照组;观察小鼠给药前的自主活动情况,然后按剂量给小鼠灌胃给药,给药后30 min和60 min记录小鼠自主活动情况.结果 给药30 min和60 min后,实验组的中枢神经作用抑制率分别为75%和60%,阳性对照组的中枢神经作用抑制率分别为100%和100%.结论 生半夏水提取液对小鼠的中枢神经的具有一定抑制作用.     

桂皮醛抗肿瘤活性及对S180荷瘤小鼠免疫功能的影响

目的:观察传统中药肉桂挥发油中桂皮醛的细胞毒作用,并以小鼠S180移植性肿瘤为模型,进行其体内外抗肿瘤活性及对S180荷瘤小鼠免疫功能影响的实验。方法:实验于2005-03/06在解放军第四军医大学药学系药物研究所实验室完成。取BALB/c小鼠60只,雌雄各半,体质量18～22g。先设1组不接种瘤株的生理盐水正常对照组,10只,雌雄各半。余50只小鼠每只右腋皮下接种0.2mL,24h后随机分成5组,即生理盐水荷瘤对照组,卡铂阳性药对照组,桂皮醛25,50,100mg/kg3个剂量组,每组10只,雌雄各半。用四甲基偶氮唑蓝法观察桂皮醛对6种人癌细胞的体外抗肿瘤作用;对25,50,100mg/kg3个剂量组分别腹腔注射相应剂量用含0.5%土温80生理盐水溶解的桂皮醛(购自中国医药集团上海化学试剂公司),正常对照组和荷瘤空白对照组腹腔注射生理盐水,阳性药对照组腹腔注射卡铂5mg/kg。接种第2天开始全部腹腔注射给药,10mL/kg,每天1次,连续给药10d,于最后1次给药后次日处死动物,测定肿瘤抑制率、免疫器官质量、血常规、NK细胞活性、T淋巴细胞转化率,分析其对小鼠体内抗肿瘤作用及与免疫调节的关系。结果:参加实验的动物60只全部进入结果分析,没有脱失。①桂皮醛对体外培养的6种人肿瘤细胞有直接细胞毒作用,其IC50的范围为12.3～37.1mg/L。②桂皮醛50,100mg/kg剂量组对S180荷瘤小鼠肿瘤生长有明显抑制作用,抑瘤率分别为33.08%和46.92%;同时能有效保护荷瘤小鼠胸腺和脾脏指数。③桂皮醛25,50mg/kg剂量组可以升高白细胞,与生理盐水荷瘤对照组比差异有显著性意义(11.13±1.49,11.25±2.18,8.78±1.33,P<0.01)。④桂皮醛25,50mg/kg剂量组的T淋巴细胞增殖能力显著或非常显著高于生理盐水荷瘤对照组(P<0.05～0.01);桂皮醛50mg/kg剂量组NK细胞杀伤活性明显高于生理盐水荷瘤对照组(P<0.05)。⑤桂皮醛100mg/kg剂量组显著降低白细胞(P<0.01);抑制T淋巴细胞增殖能力和NK细胞杀伤活性,与荷瘤空白对照组比较差异有显著性意义(P<0.01)。结论:桂皮醛对体外培养的肿瘤细胞增殖具有良好的抑制作用,在适当剂量范围内可以保护和恢复荷瘤小鼠的免疫功能。     

Antioxidant Activity of Caffeic Acid through a Novel Mechanism under UVA Irradiation

Effect of caffeic acid on the formation of hydroxyl radicals was examined during xanthone-mediated photosensitization. The reaction was performed on irradiation (λ = 365 nm) of the standard reaction mixture containing 15 µM xanthone, 0.1 M 5,5-dimethyl-1-pyrroline N-oxide (DMPO) and 20 mM phosphate buffer (pH 7.4) using electron paramagnetic resonance (EPR) with spin trapping. Caffeic acid inhibited the formation of hydroxyl radicals. Caffeic acid hardly scavenged both hydroxyl radicals and superoxide radicals under conditions employed in this paper in spite of its ability to act as a hydrogen donor or a reagent for the aromatic hydroxylation, because high concentration of DMPO trapped hydroxyl radicals overwhelmingly. Furthermore, caffeic acid inhibited the formation of hydroxyl radicals in the standard reaction mixture with EDTA under UVA irradiation. Accordingly, the inhibitory effect of caffeic acid on the formation of hydroxyl radicals in the standard reaction mixture under UVA irradiation is not due to its ability to chelate iron. Thus, the inhibitory effect of caffeic acid seems to occur in the standard reaction mixture under UVA irradiation through a novel antioxidation activity, i.e., ability to quench the exited xanthone.     

Caffeic acid inhibits the formation of 1-hydroxyethyl radical in the reaction mixture of rat liver microsomes with ethanol partly through its metal chelating activity

Effect of caffeic acid on the formation of 1-hydroxyethyl radicals via the microsomal ethanol-oxidizing system pathway was examined. The electron spin resonance spin trapping showed that 1-hydroxyethyl radicals form in the control reaction mixture which contained 0.17 M ethanol, 1 mg protein/ml rat river microsomes, 0.1 M α-(4-pyridyl-1-oxide)-N-tert-butylnitrone, 5 mM nicotinamide adenine dinucleotide phosphate and 30 mM phosphate buffer (pH 7.4). When the electron spin resonance spectra of the control reaction mixtures with caffeic acid were measured, caffeic acid inhibited the formation of 1-hydroxyethyl radicals in a concentration dependent manner. Gallic acid, dopamine, l-dopa, chlorogenic acid and catechin also inhibited the formation of 1-hydroxyethyl radicals. Above results indicated that the catechol moiety is essential to the inhibitory effect. Caffeic acid seems to chelate of iron ion at the catechol moiety. Indeed, the inhibitory effect by caffeic acid was greatly diminished in the presence of desferrioxamine, a potent iron chelator which removes iron ion in the Fe (III)-caffeic acid complex. Since Fe (III)-desferrioxamine complex is active for the 1-hydroxyethyl radicals formation, caffeic acid inhibits the formation of 1-hydroxyethyl radicals in the reaction mixture partly through its metal chelating activity.     

红豆杉茎叶镇痛作用的实验研究

目的观察红豆杉茎叶的镇痛作用。方法采用热板刺激和扭体反应实验，随机分成5组，每组小鼠10只，设立红豆杉茎叶低、中、高三个剂量组（分别相当于临床剂量0、5、10倍），并建立阿斯匹林阳性对照组和生理盐水对照组，连续口服给药3d，分别测定小鼠痛阈值及痛阈提高百分率、扭体反应潜伏期（min）、30min内扭体反应次数，并计算镇痛百分率（％）。结果红豆杉茎叶各剂量均可抑制热板刺激诱发疼痛，减少冰乙酸致痛小鼠的扭体次数，与生理盐水组比较（P〈0.05，P〈0．01），并且镇痛效应与剂量呈正相关，中、高剂量组的镇痛百分率（％）甚至高于阳性对照药物阿斯匹林组。结论红豆杉茎叶具有良好的镇痛作用。     

Effects of virgin olive oil phenolic compounds on LDL oxidation and vasorelaxation activity

This study examined the efficacy of virgin olive oil phenolic extract and other phenolic compounds (oleuropein, caffeic acid) in preventing oxidative modifications of human low density lipoprotein oxidised by CuCl2. The vasorelaxant effect of these compounds on rat aortic ring with and without functional endothelium is also discussed. Olive oil phenolic extract, caffeic acid and oleuropein increased the lag time of conjugated diene formation in a concentration-dependent manner. Moreover, phenolic extract produced a vasorelaxant effect that persisted in denuded aorta and after inhibition of nitric oxide synthase by NG-methyl-L-arginine (L-NMMA) or methylene blue. Oleuropein did not produce a relaxant effect, whereas caffeic acid produced partial relaxation at concentration 0.5 g/L.     

Antihyperglycemic and antioxidant properties of caffeic acid in db/db mice

This study investigated the blood glucose-lowering effect and antioxidant capacity of caffeic acid in C57BL/KsJ-db/db mice. Caffeic acid induced a significant reduction of the blood glucose and glycosylated hemoglobin levels than the control group. The plasma insulin, C-peptide, and leptin levels in caffeic acid group were significantly higher than those of the control group, whereas the plasma glucagon level was lower. Increased plasma insulin by caffeic acid was attributable to an antidegenerative effect on the islets. Caffeic acid also markedly increased glucokinase activity and its mRNA expression and glycogen content and simultaneously lowered glucose-6-phosphatase and phosphoenolpyruvate carboxykinase activities and their respective mRNA expressions, accompanied by a reduction in the glucose transporter 2 expression in the liver. In contrast to the hepatic glucose transporter 2, adipocyte glucose transporter 4 expression was greater than the control group. In addition, caffeic acid significantly increased superoxide dismutase, catalase, and glutathione peroxidase activities and their respective mRNA levels, while lowering the hydrogen peroxide and thiobarbituric acid reactive substances levels in the erythrocyte and liver of db/db mice. These results indicate that caffeic acid exhibits a significant potential as an antidiabetic agent by suppressing a progression of type 2 diabetic states that is suggested by an attenuation of hepatic glucose output and enhancement of adipocyte glucose uptake, insulin secretion, and antioxidant capacity.     

A water-soluble boronic acid sensor for caffeic acid based on double sites recognition

Due to reversibly and covalently binding with Lewis bases and polyols, boronic acid compounds as fluorescent sensors have been widely reported to recognize carbohydrates, ions, hydrogen peroxide, and so on. However, boronic acid sensors for highly selective recognition of caffeic acid rather than catechol or catechol derivatives have not been reported yet. Herein a novel water-soluble sensor 5c with double recognition sites based on a boronic acid was reported. When 2.3 × 10⁻⁴ M of caffeic acid was added, the fluorescence intensity of sensor 5c decreased by 99.6% via inner filter effect (IFE) because its excitation spectrum well overlaps with the absorption spectrum of caffeic acid under neutral condition, while the fluorescence increased or did not change obviously after binding with other analytes including carbohydrates and other catechol derivatives. In addition, the response time to caffeic acid is fast at room temperature, and a high binding constant (9245.7 ± 348.3 M⁻¹) and low LOD (1.81 × 10⁻⁶ M) was calculated. Moreover, determination of caffeic acid content in caffeic acid tablets was studied, and the recovery rate is sufficient. Therefore, sensor 5c can be used as a potential tool for detecting biologically significant caffeic acid in real samples.

Herein, the specific recognition of caffeic acid by the double sites boronic acid sensor 5c is reported. The synergistic effect of the two recognition sites greatly improves the binding affinity and selectivity of the sensor.     

Identification of an ascorbic acid metabolite among "uremic middle molecules"

Among uremic toxins in the middle molecular mass range, 1H, 13C-nuclear magnetic resonance, ultraviolet spectrometry, and chromatographic analyses allow identification of the main component of the so-called "2-5-3 fraction" as ascorbic acid 2-sulfate, a conjugated metabolite of ascorbic acid. We previously (Clin Nephrol 1986;25:212-8) showed an inhibitory effect of the 2-5-3 fraction on microtubule formation. Therefore, we tested the action of ascorbic acid 2-sulfate and its synthetized enantiomers on tubulin polymerization. Because these molecules did not exert any inhibitory effect, we hypothesize that the 2-5-3 fraction is a mixture of compounds in which only a very low quantity of the inhibitory factor is present.     

Failure of ischemic neuroprotection by potentiators of gamma-aminobutyric acid

INTRODUCTION: Potentiators of inhibitory neurotransmission may provide a neuroprotective effect on cerebral tissue exposed to ischemia, without inducing toxic side effects. Topiramate and vigabatrin enhance the action of gamma-aminobutyric acid (GABA), and each has side effect profiles known to be well tolerated through their clinical use as anticonvulsant medications. We assessed the potential benefit through GABA activation by these drugs on infarct size and functional recovery following focal cerebral ischemia in mice. METHODS: Silicon-coated suture was advanced through the internal carotid artery of 89 halothane-anesthetized mice to temporarily occlude the right middle cerebral artery for either 45 minutes (topiramate), or 120 minutes (vigabatrin). Animals were treated either at the time of reperfusion with topiramate (100 mg/kg, 40 mg/kg, or saline control), or two hours before arterial occlusion with vigabatrin, (1000 mg/kg, 500 mg/kg, or saline control). Neurological outcome was measured 24 hours after ischemia using a 28-point functional examination score. Infarct volume was estimated by summing area maps of stained slices of infarcted hemispheres. RESULTS: Functional examination scores at 24 hours were similar between the high dose topiramate group, the low dose topiramate group, and the control group. Similarly, no differences were noted between examination scores of high dose vigabatrin, low dose vigabatrin, and control. Consistent sized right hemisphere infarcts were noted within each group on histological examination. Mean infarct volumes did not differ between groups treated with high dose topiramate, low dose topiramate, or control. Infarct volumes of animals treated with saline control were slightly larger than that of high dose vigabatrin and low dose vigabatrin groups, but the difference did not reach significance. CONCLUSION: Treatment with these two potentiators of GABA did not result in significant differences in outcome following focal cerebral ischemia, by either functional or histological measures. These results do not support a substantial neuroprotective role of GABA following ischemia in this mouse suture model.     

盐酸戊乙奎醚对内毒素休克兔血清肿瘤坏死因子-α、一氧化氮、内皮素-1的影响

目的:观察不同剂量盐酸戊乙奎醚(长托宁)对内毒素休克兔血清肿瘤坏死因子-α(TNF-α)、一氧化氮(NO)、内皮素-1(ET-1)浓度的影响.方法:35只健康新西兰大白兔随机分成正常对照组、休克组、长托宁低剂量组(0.05 mg/kg)、长托宁中剂量组(0.15 mg/kg)和长托宁高剂量组(0.45 mg/kg),每组7只.采用静脉注射大肠杆菌脂多糖制作内毒素休克模型.长托宁各组分别于静注脂多糖后立即静注不同剂量长托宁,测定给药后不同时间点动物血清TNF-α、NO、ET-1的含量.结果:休克组血清TNF-α、NO、ET-1浓度明显高于正常对照组(P＜0.01);长托宁低、中、高剂量组血清TNF-α、NO、ET-1浓度较休克组均有不同程度减低(P＜0.05),以中剂量组减低最明显(P＜0.01或P＜0.05).结论:长托宁能部分抑制炎症介质的产生和释放,相对于低、高剂量长托宁,中剂量在抑制炎症介质方面效果更为显著.