基于Notch信号通路探讨β-榄香烯逆转肺腺癌耐药机制的研究

目的基于Notch信号通路,探讨β-榄香烯逆转肺腺癌耐药的机制。方法选择对A549/DDP细胞无细胞毒性的β-榄香烯浓度处理A549/DDP细胞,设β-榄香烯组与对照组,两组细胞均给予不同浓度的DDP(0.25、0.5、1、2、4、8、16、32μg·mL^-1)处理24h,MTT法检测细胞活力并计算IC₅₀,流式细胞仪检测细胞凋亡,Western blotting检测凋亡相关蛋白(cleaved-caspase 3、cleaved-caspase 9)、耐药相关蛋白(P-gp、survivin、p21)及Notch信号通路蛋白(Notch1、Hes1、Hey2)的表达。A549/DDP细胞在β-榄香烯处理的基础上,联合1μg·mL^-1Notch信号通路激活剂rhNF-κB处理,比较两组的IC₅₀,Western blotting检测P-gp以及Notch1、Hes1、Hey2的表达。结果当β-榄香烯浓度低于20μg·mL^-1时,其对A549/DDP细胞的抑制率小于10%,无细胞毒性。β-榄香烯组DDP的IC₅₀显著低于对照组(t=14.712,P<0.05),逆转倍数为4.452倍。β-榄香烯组细胞凋亡率以及凋亡相关蛋白cleavedcaspase 3和cleaved-caspase 9的表达水平均高于对照组,差异有统计学意义(P<0.05)。β-榄香烯组P-gp、survivin蛋白相对表达量低于对照组,而p21蛋白相对表达量高于对照组,差异均有统计学意义(P<0.05)。β-榄香烯组Notch1、Hes1、Hey2蛋白相对表达量均低于对照组,差异有统计学意义(P<0.05)。β-榄香烯+rhNF-κB组DDP的IC₅₀以及P-gp、Notch1、Hes1、Hey2蛋白表达水平均高于β-榄香烯组,差异有统计学意义(P<0.05)。结论β-榄香烯可通过抑制Notch信号通路促进A549/DDP细胞凋亡,并逆转肺腺癌耐药。     

榄香烯乳胸腔内灌注治疗晚期肺癌伴胸水临床观察

目的：比较榄香烯乳（治疗组）与顺铂（对照组）胸腔内注射对恶性胸水的疗效及毒性。方法：治疗组30例采用抗肿瘤新药榄香烯乳500mg胸腔内灌注7－10天1次，用药2－3次，对照组20例胸腔内灌注顺铂80mg 7-10天1次，用药2－3次。结果：治疗组有效率（CR＋PR）83．33％，对照组有效率85％，两组有效率差异无显著性（P＞0．05），胃肠道及血液学毒性反应治疗上明显低于对照组（P＜0．01）。结论：榄香烯乳胸腔内灌注是一种治疗癌性胸水的有效新药，且无毒副反应。     

榄香烯联合5-FU体内外抑制人胃癌MGC80-3细胞增殖与诱导凋亡作用的研究

目的 研究中药莪术提取物榄香烯制剂(Elemene)联合5-氟尿嘧啶(5-FU)对MGC80-3细胞增殖的抑制及诱导凋亡的作用。方法 采用MTT法考察榄香烯、5-FU单药及联合作用于MGC80-3细胞的体外抗肿瘤效应;应用中效原理进行统计分析单药及联合用药对肿瘤细胞生长的影响,确定联合用药时的效应(Fa)合用指数(CI)的关系;流式细胞仪检测细胞凋亡周期改变。通过建立MGC80-3裸鼠体内异种肿瘤模型,考察榄香烯及5-FU单用及合用对荷瘤小鼠肿瘤生长的抑制作用。结果体外试验结果表明榄香烯与5-FU联合应用作用于MGC80-3细胞可产生较好协同效应,单染法(PI)检测到榄香烯与5-FU联合之后能够使MGC80-3细胞表现出显著的G0/G1期阻滞作用;体内试验结果表明榄香烯单药及与5-FU联合作用对MGC80-3裸鼠异种肿瘤生长均有明显的抑制作用,且联合用药组抑瘤率可达84.68%。结论 榄香烯联合5-FU作用于MGC80-3肿瘤细胞株可产生较好的协同效应。     

榄香烯乳、顺铂治疗恶性腹水疗效的对照观察

目的：评价榄香烯治疗恶性腹水的临床疗效。方法：８７ 例恶性腹水患者随机分为两组,榄香烯组４０ 例,顺铂组４７ 例。两组均采用腹腔内注射药物治疗。结果：榄香烯组总有效率为８５ ％ ,明显高于顺铂组的７０ ．１８ ％ , Ｐ＜ ０ ．０５ 。两组不良反应发生率分别是２２ ．５ ％ 及５１ ．０６ ％ ,榄香烯组明显低于顺铂组,差异具有显著性, Ｐ＜ ０ ．０５ 。榄香烯治疗后患者 Ｃ Ｄ４ 细胞数量明显增加 Ｃ Ｄ４／ Ｃ Ｄ８ 亦明显升高。结论：榄香烯能有效地控制恶性腹水,不良反应少,有一定改善患者免疫功能作用,值得推广应用。     

榄香烯诱导肺癌A549细胞凋亡作用的剂量和时间依赖性研究

目的探讨榄香烯对肺癌A549细胞的增殖抑制作用及其对细胞凋亡的影响。方法每批细胞按空白对照组和药物处理组随机分组,检测榄香烯对癌细胞的增殖抑制作用;采用流式细胞光度分析术(FCM)检测细胞凋亡率及细胞周期时相分布,同时进行细胞的形态学观察。结果榄香烯存在剂量依赖性和时间依赖性,榄香烯在48 h、72 h对A549细胞周期有明显影响;榄香烯可阻滞A549细胞于S期并诱导细胞凋亡,但细胞凋亡率并不完全与药物浓度成正相关。结论榄香烯对肿瘤细胞具有较强的增殖抑制作用,在一定药物浓度范围内,随药物浓度的增加榄香烯可使细胞凋亡率升高,但超过一定作用浓度,药物的细胞毒作用增强,细胞凋亡率则呈下降趋势。     

榄香烯联合氟尿嘧啶对肝癌H22荷瘤小鼠的抑瘤作用

目的观察榄香烯联合氟尿嘧啶（5-FU）对H22荷瘤小鼠的抑瘤作用。方法小鼠左腋窝下皮下种瘤造实体瘤模型。将35只荷瘤小鼠等分为5组,分别以榄香烯高剂量（100mg·kg^-1）、榄香烯低剂量（50mg·kg^-1）、5．FU（20mg·kg^-1）、榄香烯低剂量（50mg·kg^-1）联合5．FU（20mg·k^-1）及生理盐水（0．2mE）处置10d,记录各组小鼠治疗期间体质量及移植瘤的体积并计算抑制率,HE染色法观察各切片中瘤组织的差异。结果荷瘤小鼠经药物处置5d后,各用药组瘤体积较生理盐水对照组均有缩小,差异有统计学意义（P〈0．05）,联合组优于单独用药组;对小鼠体质量影响,联合组及5-FU组均大于榄香烯组及对照组;组织切片中可见各用药组对瘤组织不同程度的破坏。结论榄香烯能有效地抑制H22肿瘤的生长,可以作为化疗药5-Fu的辅助药物。     

榄香烯乳联合5-氟尿嘧啶对人胃癌细胞AGS增殖的抑制作用研究

目的:研究榄香烯乳联合5-氟尿嘧啶(5-FU)对人胃癌细胞AGS增殖的抑制作用。方法:取对数生长期AGS细胞,分为榄香烯乳各剂量[0(对照组)、20、40、60、80μg/ml]组、5-FU组(40μg/ml)及其混合组(榄香烯乳80μg/ml+5-FU 40μg/ml),每个浓度3个复孔,采用MTT法考察榄香烯乳各剂量组作用12、24、48 h,5-FU组和混合组作用24 h后细胞的增殖抑制率;采用流式细胞仪、蛋白质印迹法检测对照组、榄香烯乳组(80μg/ml)、5-FU组和混合组作用24 h的细胞周期和细胞周期蛋白D1(Cyclin D1)的表达情况。以接种AGS细胞的裸鼠为模型,考察模型组、5-FU组[17 mg/(kg·d)]、榄香烯乳组[75 mg/(kg·d)]及其混合组荷瘤鼠体内的抑瘤率。结果:榄香烯乳能呈浓度依赖性抑制细胞增殖(P<0.05);混合组对细胞的抑制作用强于二者单用组;与对照组比较,榄香烯乳组、5-FU组和混合组细胞G0/G1期比例增加,差异具有统计学意义(P<0.05);后3组细胞中Cyclin D1表达依次减弱,差异具有统计学意义(P<0.05)。与模型组比较,榄香烯乳组、5-FU组和混合组的体内抑瘤率分别为45.16%、52.68%、65.59%。结论:榄香烯乳与5-FU具有协同抑制AGS细胞增殖的作用。     

中药榄香烯在宫颈癌新辅助化疗中的应用研究

目的研究榄香烯在宫颈癌新辅助化疗中的作用。方法建立宫颈癌U14小鼠模型,将60只小鼠分为对照组、榄香烯组、PF组、榄香烯＋PF组,于治疗后第15天取小鼠血清及肿瘤组织,通过流式细胞仪检测肿瘤细胞的凋亡情况,检测血清中T细胞亚群的变化。结果榄香烯能促进肿瘤细胞凋亡,与对照组比较差异有统计学意义（P〈0.05）;且能提高荷瘤鼠的免疫功能,与对照组比较差异有统计学意义（P〈0.05）。结论榄香烯可用于宫颈癌新辅助化疗。     

全脑放疗联合榄香烯治疗非小细胞肺癌脑转移瘤的临床效果

目的观察全脑放疗联合榄香烯治疗非小细胞肺癌脑转移瘤的临床效果。方法收集2015年1月-2016年12月广州医科大学附属肿瘤医院收治的非小细胞肺癌脑转移瘤患者58例,所有患者均接受全脑放疗,其中接受榄香烯治疗的患者23例为观察组,未接受榄香烯治疗的患者35例为对照组。比较2组临床疗效,治疗前后血清癌胚抗原(CEA)、鳞状细胞癌抗原(SCC)水平,以及生存率。结果观察组总有效率(95.65%)高于对照组(77.14%)(χ²=14.575,P=0.000);治疗后,2组血清CEA和SCC水平均低于治疗前,且观察组低于对照组(P<0.01);2组患者1、2、3年生存率比较均无显著差异(P>0.05)。结论全脑放疗联合榄香烯治疗非小细胞肺癌脑转移瘤的效果较好,可显著降低患者血清CEA和SCC水平,抑制肿瘤进展,值得临床应用。     

榄香烯对白血病K562细胞中凋亡相关基因PDCD5表达的影响

目的探讨榄香烯对白血病K562细胞中凋亡相关基因PDCD5表达的影响。方法分别运用不同浓度榄香烯处理白血病K562细胞,并采用免疫细胞化学SP法检测PDCD5蛋白的表达情况。结果不同浓度的榄香烯均可上调白血病K562细胞中PDCD5蛋白表达,榄香烯作用组与对照组两两相比,差异均有统计学意义(P均<0.05)。结论榄香烯可通过上调白血病K562细胞中PDCD5蛋白表达,从而诱导肿瘤细胞凋亡。     

龙眼参提取物对环磷酰胺所致小鼠免疫功能低下的影响

目的 :探讨龙眼参提取物对环磷酰胺所致小鼠免疫功能低下的影响。方法 :环磷酰胺诱导小鼠免疫功能低下 ,口服给予小鼠不同剂量的龙眼参提取物30d。结果 :龙眼参提取物大、小剂量组均能提高中性粒细胞杀伤功能 (P<0 05 ,P<0 01)及小鼠溶血素水平 (P<0 05) ,同时还均具有提高碳粒廓清指数和吞噬指数的作用。结论 :龙眼参提取物对小鼠的免疫功能有较好的改善作用。     

姜黄素通过阻断NF-κB减少IL-1β诱导的内皮脂酶表达

目的观察姜黄素对培养的人血管内皮细胞内皮脂酶表达的影响,并探讨其可能的作用机制。方法不同浓度的姜黄素处理HUVEC-12细胞。RT-PCR检测内皮酯酶(endo-thelial lipase,EL)mRNA的表达;Western blot检测核因子-κB抑制因子-α(inhibitor of nuclear factor-κB-α,IκB-α)蛋白的表达;间接免疫荧光检测核因子-κB(nuclear factor-κB,NF-κB)蛋白的活化。结果IL-1β处理HUVEC-12细胞可以明显上调EL mRNA的表达,同时降低胞质蛋白IκB-α的表达水平,激活核转录因子NF-κB,增加胞核蛋白NF-κB的水平。姜黄素预处理HUVEC-12细胞可以抑制IL-1β对EL的上调作用,同时逆转IL-1β诱导的IκB-α蛋白降解和NF-κB活化。结论姜黄素可以通过阻断NF-κB活化减少IL-1β诱导的人血管内皮细胞EL的表达。     

Effect of Eurycoma longifolia stem extract on uric acid excretion in hyperuricemia mice

OBJECTIVE Eurycoma longifolia is a tropical medicinal plant belonging to Simaroubaceae distributed in South East Asia. The aim of this study is to explore the effect and mechanism of E. longifolia stem 70% ethanol extract(EL) and its active compoundson uric acid excretion. METHODS Potassium oxonate(PO) induced hyperuricemia rats and adenine-PO induced hyperuricemia mouse model were used to evaluate the effects of EL. Ultra Performance Liquid Chromatography was used to determine the levels of plasma or serum uric acid and creatinine. Hematoxylin-eosin staining was applied to observe kidney pathological changes, Western blotting was applied to detect protein expression levels of uric acid transporters. Effects of constituents on urate uptake were tested in h URAT1-expressing HEK293 T cells. RESULTS EL significantly reduced serum and plasma uric acid levels at dosages of 100, 200 and400 mg·kg~(-1) in hyperuricemia rats and mice, and increased the clearance rate of uric acid and creatinine, improved therenal pathological injury. The protein expression levels of urate reabsorption transporter 1(URAT1) and glucose transporter 9 were down-regulated while sodium-dependent phosphate transporter 1 and ATP-binding cassette transporter G2 were up-regulated in the kidney after EL treatment. The diterpenes(50 μmol·L~(-1)) isolated from EL showed inhibitory effects on urate uptake in h URAT1-expressing HEK293 T cells,and the effect of eurycomanol was further confirmed in vivo. CONCLUSION EL significantly reduced blood uric acid levels and prevented pathological changes of kidney in PO induced hyperuricemia animal model, improved renal urate transports. We partly clarified the mechanism was related to suppressing effect of URAT1 by diterpene in EL. This study is the first to demonstrate that EL plays a role in hyperuricemia by promoting renal uric acid excretion.     

Effect of Bile Duct Ligation and Unilateral Nephrectomy on Brain Concentration and Convulsant Potential of the Quinolone Antibacterial Agent Levofloxacin in Rats

To mimic the excretion route of the quinolone antibacterialagent levofloxacin (LVFX) in humans, we produced an excretion-limited(EL) model in male Sprague–Dawley rats by bile duct ligationand unilateral nephrectomy. We then examined the relationshipbetween brain levels of LVFX and its convulsant effects in controland EL animals. Serum concentrations of LVFX in EL animals (EL+ LVFX) were 2.38- and 1.59-fold and brain concentrations were1.33- and 1.19-fold those of the controls (control + LVFX) at30 min after a single intravenous injection of 10 and 100 mg/kgLVFX, respectively. Furthermore EL animals became more susceptibleto the convulsant effect of LVFX with a 1.28-fold decrease inconvulsion-inducing dose. In combination with oral pretreatmentwith 400 mg/kg 4-biphenylacetic acid (BPAA), convulsion-inducingdoses in the control (control + LVFX + BPAA) and EL (EL + LVFX+ BPAA) groups were markedly decreased by 2.25 and 9 times thatof the control + LVFX group. EL operation and BPAA pretreatmentslowed the elimination of LVFX in the serum and brain 4 hr laterin the following order: EL + LVFX + BPAA, control + LVFX + BPAA,EL + LVFX, and control + LVFX groups. This order reflects thatfor the convulsion-inducing doses. These results suggest thatEL rats may be a useful model for humans and that the convulsanteffect of LVFX with or without BPAA arises not only from theattainment of maximum brain concentration but also from delayeddisappearance from the brain.     

Non-ionic surfactants as novel intranasal absorption enhancers: in vitro and in vivo characterization

Objective: To explore the potential of non-ionic surfactants as novel intranasal absorption enhancers.

Methods: Taking sumatriptan succinate (SMS) as a model drug, influence of different non-ionic surfactants, including laurate sucrose ester (SE), cremophor EL and poloxamer 188, on the intranasal absorption of SMS was investigated using an in situ nasal perfusion technique in rats. Ciliotoxicity of the non-ionic surfactants was evaluated using an in situ toad palate model. In vivo behavior of the selected formulations was studied in rats.

Results: All the non-ionic surfactants investigated increased the intranasal absorption of SMS remarkably but with varied extent and trend. Moreover, it was revealed that at the same concentration, laurate SE had better permeation-enhancing effect than that of cremophor EL and poloxamer 188. The ciliotoxicity results showed that all the non-ionic surfactants were regarded as safe at selected concentrations. Based on the in situ absorption data and ciliotoxicity results, the following three samples, 0.5% laurate SE, 0.1% cremophor EL and 0.5% poloxamer 188 were selected for in vivo absorption studies in rats. Among them, 0.5% laurate SE group presented the highest enhancing effect, followed by 0.1% cremophor EL and 0.5% poloxamer 188 group, with absolute bioavailability 29.99%, 22.64% and 20.90%, respectively.

Conclusions: Laurate SE is a promising intranasal absorption enhancer.     

Subchronic Oral Toxicity of Triethyl Lead in the Male Weanling Rat. Clinical, Biochemical, Hematological, and Histopathological Effects

Subchronic Oral Toxicity of Triethyl Lead in the Male WeanlingRat. Clinical, Biochemical, Hematological, and HistopathologicalEffects. YAGMINAS, A. P., FRANKLIN, C. A., VILLENEUVE, D. C,GILMAN, A. P., LITTLE, P. B., AND VALLI, V. E. O. (1990). Fundam.Appl. Toxicol. 15, 580–596. This study was designed toascertain the effects of low level exposure of triethyl lead(3EL) to the male weanling rat. Groups of 20 animals were administeredby gavage 3EL at 0.05, 0.10, 0.20, 0.50, and 1.00 mg/kg bodywt for 91 days, 5 days/week. Lead acetate (PbHOAC) at 200 mg/kgbody wt/day was given as a positive control. Weight gain wasreduced in those animals receiving 1.0 3EL. Spleen and kidneyweights were elevated in the PbHOAC group. Residues of 3EL andits metabolites diethyl lead (2EL) and lead (Pb) accumulatedin a dose-dependent manner in blood, liver, kidney, and brain;3EL accumulated preferentially in the liver while inorganiclead accumulated in the kidney. Dose-dependent changes occurredin serum calcium which was decreased and in phosphorus whichwas elevated for all dose groups. Serum cholesterol was elevatedin the three highest 3EL groups as was alkaline phosphatase.LDH was lowered in the PbHOAC-treated group but microsomal anilinehydroxylase was elevated. Hematological changes consisted ofelevated platelet counts in the 1.0 3EL group and decreasedmean corpuscular hemoglobin content and mean corpuscular volumein the PbHOAC-treated group. Treatment related histopathologicalchanges were seen in thyroid, liver, kidney, and bone marrow.Based on these data a no observed adverse effect level for 3ELwas at 0.10 mg/kg/body wt.     

Influence of enteric-coated lactose on the release profile of 4-aminopyridine from HPMC matrix tablets

A weakly basic experimental drug, 4-aminopyridine, was taken as a model to study the influence of enteric-coated lactose (EL) on the release profile from hydroxypropyl methylcellulose matrices. Powder mixtures were wet-granulated with water. The dried granulation was compressed with a hydraulic press at 85 MPa. Dissolution studies were made using HCl 0.1 N and then phosphate buffer pH 7.4. Dissolution curves were described by M(t)/M(inf) = k*t(N). A trend toward increasing exponent (n) and decreasing release constant (k) values is observed with increasing EL concentrations up to 9%; this is attributed to an increasing obstruction of the diffusion path by isolated EL particles that are insoluble in HCl and are surrounded by a water-filled space. After a critical EL concentration, the water-filled spaces surrounding EL particles percolate, producing the opposite effect, increasing the release constant and decreasing the exponent (n) values as the EL proportion increases from 10% to 50%. EL particles (2% to 9%) decrease the drug and water transport in matrices dissolving in HCl. Thereafter, at pH 7.4, the pores formed by dissolution of EL particles produce the opposite. Both processes contribute to flattening the release profile. Release profiles with decreasing release constant values show a logarithmic trend toward increasing values of the exponent (n), changing from diffusion toward relaxation-erosion-controlled processes.     

Functional dissociation of striatal and hippocampal cholinergic systems in egocentric and allocentric localization: effect of overtraining.

The effects of overtraining on the retention of egocentric localization (EL) and allocentric localization (AL) tasks using rats injected with 1.8 nmol/12 microliters of ethylcholine mustard aziridinium ion (AF64A) in the striatum or hippocampus were examined. In the EL task, an arm positioning at in same direction of a randomly selected start arm was baited throughout trials. In the AL task, a baited arm was fixed throughout trials regardless of the position of the start arm. The performance of EL retention was disrupted only by intrastriatal AF64A injection. The overtrained striatal lesioned animals showed better performance compared to the non-overtrained striatal lesioned animals. In contrast, hippocampal AF64A injection selectively impaired AL retention. Unlike the effect of overtraining on EL retention in striatal lesioned animals, however, the AL performance of hippocampal lesioned animals was not significantly affected by overtraining. The results support the idea of functional dissociation of striatal and hippocampal cholinergic systems in EL and AL learning and provide further evidence that the striatal cholinergic system may be involved in the memory (consolidation or retrieval) process of EL learning, and the hippocampal cholinergic system in the processing of specific cues necessary for efficient performance in AL learning rather than memory.     

Systemic treatment with n-6 polyunsaturated fatty acids attenuates EL4 thymoma growth and metastasis through enhancing specific and non-specific anti-tumor cytolytic activities and production of TH1 cytokines

Recently, there has been a great interest in the effects of different types of n-6 polyunsaturated acids (n-6 PUFAs) upon the immune system and cancer development. However, the effects of n-6 PUFAs are still controversial and as yet undefined. The present study aimed to investigate the anti-tumor effects of n-6 PUFAs against EL4 thymoma and the associated immune mechanisms. To this, sesame oil, a vegetable oil enriched with n-6 PUFAs, or free linoleic acid (LA) were administered intraperitoneally into C57BL/6 mice before and after challenge with EL4 lymphoma cells. Treatment with either sesame oil or LA attenuated the growth and metastasis of EL4 lymphoma. The anti-tumor effect of LA was superior to that of sesame oil, and associated with an increase in the survival rate of the tumor-bearing mice. In addition, both sesame oil and LA showed dose-dependent anti-lymphoma growth in vitro. Treatment with LA generated significant increases in the anti-lymphoma cytolytic and cytostatic activities of T cells and macrophages, respectively, and enhanced production of IL-2 and IFN-gamma while decreased production of IL-4, IL-6 and IL-10. In summation, the results suggest that n-6 PUFAs, represented by LA, can attenuate EL4 lymphoma growth and metastasis through enhancing the specific and non-specific anti-tumor cytolytic activities and production of TH1 cytokines. These findings might be of great importance for a proper design of systemic nourishment with PUFAs emulsions for cancer patients.     

Preclinical evaluation of alternative pharmaceutical delivery vehicles for paclitaxel

New solubilizers, including Sorporol 230, Sorporol 120Ex, Aceporol 345-T, Aceporol 460 and Riciporol 335, as potential new delivery vehicles for paclitaxel were investigated, since recent studies have shown that the paclitaxel delivery vehicle Cremophor EL significantly alters the pharmacokinetics of paclitaxel. Cremophor EL and Tween 80 were used as a reference. As in the case of Cremophor EL, alteration of blood distribution of paclitaxel occurred in the presence of all tested vehicles. Also, no differences in the affinity of paclitaxel for the tested solubilizers was found during equilibrium dialysis experiments. The different vehicles could be distinguished by a different rate of esterase-mediated breakdown, which was correlated with the fatty acid content of the solubilizers. The activation of the complement cascade was less pronounced for all solubilizers, except Riciporol 335, compared to Cremophor EL. The strategies presented here provide the possibility to rapidly screen future candidate delivery vehicles with optimal characteristics for use as a solubilizer in clinical formulations of paclitaxel or other poorly water-soluble drugs.