Antrodia camphorata inhibits proliferation of human breast cancer cells in vitro and in vivo

Antrodia camphorata (A. camphorata) has been shown to induce apoptosis in cultured human breast cancer cells (MDA-MB-231). In this study, we report the effectiveness of the fermented culture broth of A. camphorata in terms of tumor regression as determined using both in vitro cell culture and in vivo athymic nude mice models of breast cancer. We found that the A. camphorata treatment decreased the proliferation of MDA-MB-231 cells by arresting progression through the G1 phase of the cell cycle. This cell cycle blockade was associated with reductions in cyclin D1, cyclin E, CDK4, cyclin A, and proliferating cell nuclear antigen (PCNA), and increased CDK inhibitor p27/KIP and p21/WAF1 in a dose and time-dependent manner. Furthermore, the A. camphorata treatment was effective in delaying tumor incidence in the nude mice inoculated with MDA-MB-231 cells as well as reducing the tumor burden when compared to controls. A. camphorata treatment also inhibited proliferation (cyclin D1 and PCNA) and induced apoptosis (Bcl-2 and TUNEL) when the tumor tissue sections were examined histologically and immunohistochemically. These results suggest that the A. camphorata treatment induced cell cycle arrest and apoptosis of human breast cancer cells both in vitro and in vivo.     

咖啡酸苯乙酯纳米混悬剂的制备及其体外抑制乳腺癌细胞作用研究

目的 优化咖啡酸苯乙酯纳米混悬剂的处方,并考察其体外抑制乳腺癌细胞作用.方法 以泊洛沙姆188、蜂胶作为载体,采用反溶剂沉淀法制备,通过星点设计–效应面法优化最佳制备工艺参数,并考察咖啡酸苯乙酯纳米混悬剂的稳定性、载药量、包封率以及冻干保护剂的筛选,同时考察对4T1乳腺癌细胞的生长抑制作用和细胞摄取情况.结果 咖啡酸苯...     

Protective effect of ferulic acid on cisplatin induced nephrotoxicity in rats

This study aims to determine the potential protective effects of ferulic acid against cisplatin-induced nephrotoxicity and to compare its effect with curcumin, a well-known protective agent against cisplatin- induced toxicity in rats. Administration of cisplatin resulted in high BUN (Blood Urea Nitrogen), creatinine, MDA (Malondialdehyde), MPO (Myeloperoxidase), TOS (Total Oxidative Status), PtNT (Protein Nitrotyrosine) levels (p < 0.05). Histological observations showed abnormal morphology of kidney; in addition with appearance of TUNEL positive cells indicating apoptosis in cisplatin administered group. HO-1 (Heme Oxygenase-1) levels measured by RT-PCR (Real Time Polymerase Chain Reaction), and TAS (Total Antioxidative Status) revealed antioxidant depletion due to cisplatin toxicity in animals (p < 0.05). All parameters showed improvement in groups treated with ferulic acid (p < 0.05). Ferulic acid treatment was found significant in preventing oxidative stress, increasing antioxidative status and regaining histological parameters to normal, indicating nephroprotective and antioxidant effects of this phenolic compound.     

去甲二氢愈创木酸类似物26C基于活性氧簇诱导细胞凋亡的体外抗肺癌活性研究

目的 研究去甲二氢愈创木酸（NDGA）类似物26C的抗肺癌活性与初步机制。方法 利用MTT实验评价26C对肺癌细胞NCI-H460的细胞毒性;用集落克隆、划痕实验分别检测26C对NCI-H460细胞生长、迁移的影响;流式细胞仪检测26C对NCI-H460细胞的周期阻滞及诱导凋亡情况;活性氧簇（ROS）实验探究26C引起细胞凋亡的作用机制。结果 26C对NCI-H460的IC₅₀为（4.7±0.5）μmol/L,对NCI-H460的集落形成、迁移有较强的抑制作用,其活性明显优于先导化合物NDGA;26C可将细胞周期阻滞在G₂/M期,并通过升高ROS水平的作用机制诱导NCI-H460细胞凋亡。结论 化合物26C为具有研发前景的抗肺癌候选化合物,其通过提高ROS水平、阻滞细胞周期来抑制细胞生长和诱导细胞凋亡。     

路路通酸对乳腺癌MCF-7细胞和宫颈癌C-33A细胞增殖的影响

目的研究路路通酸(BTA)对乳腺癌MCF-7细胞及宫颈癌C-33A细胞增殖的影响。方法采用MTT法检测不同浓度BTA作用不同时间后对MCF-7细胞、C-33A细胞的增殖抑制作用;流式细胞仪检测BTA处理后细胞周期的变化。结果 BTA作用于乳腺癌MCF-7细胞24、48、72 h后的IC50分别为(37.62±1.72)、(27.32±0.99)、(19.19±0.90)μmol/L。BTA作用于宫颈癌C-33A细胞24、48、72 h后的IC50分别为(34.55±0.88)、(27.20±1.03)、(16.74±0.79)μmol/L,BTA对2种细胞增殖有明显抑制作用,且呈浓度时间依赖性(P<0.05),流式结果显示,BTA将MCF-7细胞阻滞在S期,并诱导其凋亡;BTA将C-33A细胞阻滞在G1-S期。结论BTA对乳腺癌MCF-7细胞和宫颈癌C-33A细胞具有较强的增殖抑制作用,其机制与细胞周期阻滞和诱导细胞凋亡有关。     

Butyrate reduces the frequency of micronuclei in human colon carcinoma cells in vitro

Butyrate, formed by bacterial fermentation of plant foods, has been shown to protect human colon cells from selected genotoxic substances. The mechanism for this effect could be the enhancement of toxicological defence leading to an increased detoxification of genotoxic risk factors and thus to a reduction of DNA and chromosome damage. Previous protective properties of butyrate against DNA damage induction in colon cells were demonstrated using the comet assay. In the present study the effect of butyrate on chromosome damage induced by ferric nitrilotriacetate (Fe-NTA) and hydrogen peroxide (H₂O₂) (suggested to be putative risk factors of colorectal carcinogenesis) was investigated using the cytokinesis-block micronucleus (CBMN) test. It was possible to reveal that pre-treatment of HT29 colon carcinoma cells with butyrate (2 and 4 mM) for 15 min caused a reduction of micronuclei induced with H₂O₂ (75 μM; p < 0.01) and Fe-NTA (500 and 1000 μM; p < 0.05). The decrease in the level of Fe-NTA- and H₂O₂-induced micronuclei was also confirmed in most of the corresponding variants of 24 h pre-treatment of cells with butyrate. The results obtained demonstrate for the first time protective properties of butyrate against chromosome damage induced by H₂O₂ and Fe-NTA in human colon carcinoma cells.     

新疆阿魏树脂不同极性部位对人肺癌细胞的体外抑制作用研究

目的研究新疆阿魏的提取物、不同极性部位以及倍半萜香豆素类化合物对人肺癌细胞的体外抑制作用,明确新疆阿魏树脂的活性成分。方法应用MTT法检测新疆阿魏树脂提取物(100 mg/m L)、不同极性部位(100 mg/m L)和倍半萜香豆素化合物(100、50、10、1、0.1μmol/L)对人肺癌细胞系A549、H292、H1792、H460作用48 h后生长的抑制作用,采用抑制率和IC50为指标衡量其活性。结果新疆阿魏树脂氯仿提取物在100 mg/m L质量浓度下对A549、H292、H1792、H460细胞系的抑制率分别为75.45%、65.39%、46.54%、73.68。低极性部位在100 mg/m L质量浓度下对4种细胞系的生长抑制率分别为88.74%、80.62%、60.11%、92.35%。从低极性部位中分离得到的主要倍半萜香豆素类化合物farnesiferol B、farnesiferone A、farnesiferol C、多花素宁,其中farnesiferol C、多花素宁的IC50分别为35.49、15.44、44.25、21.63μmol/L和23.60、17.90、38.33、12.35μmol/L,表现出显著的生长抑制作用,且具有剂量相关性。结论新疆阿魏树脂的低极性部位为体外抑制人肺癌细胞生长作用的活性部位,其所含有的高含量倍半萜香豆素化合物farnesiferol C为其主要活性成分。     

胃肠安联合替加氟对结肠癌HT29细胞体内外抑制作用的研究

目的研究胃肠安联合替加氟对结肠癌HT29细胞的体内外抑制作用。方法采用MTT法测定胃肠安联合替加氟对结肠癌HT29细胞的抑制作用,并采用流式细胞仪检测结肠癌HT29细胞中CD133+的表达情况。建立结肠癌HT29细胞裸鼠模型,对照组尾静脉注射生理盐水,0.2 mL/次,2次/周。替加氟组尾iv替加氟注射液,36 mg/kg,1次/周;胃肠安丸组,用纯净水将按胃肠安丸配成4 mg/kg,ig 0.2 mL/d;胃肠安+替加氟组尾iv替加氟注射液,36 mg/kg,1次/周;同时,用纯净水将按胃肠安丸配成4 mg/kg,ig 0.2 mL/d。观察胃肠安联合替加氟对荷瘤裸鼠生存状况和结肠癌HT-29细胞的影响。结果与对照组比较,替加氟组、胃肠安组、胃肠安+替加氟组抑制结肠癌HT-29细胞的增殖能力明升高,差异有统计学意义(P0.05);胃肠安+替加氟组明显高于单独使用替加氟组、胃肠安组,差异有统计学意义(P0.05)。与对照组比较,替加氟组、胃肠安组、胃肠安+替加氟组的CD133+细胞比例明显降低,差异有统计学意义(P0.05);胃肠安+替加氟组明显低于单独使用替加氟组、胃肠安组,差异有统计学意义(P0.05)。治疗后,胃肠安组、替加氟组、胃肠安+替加氟组的实体瘤质量明显低于对照组,差异具有统计学意义(P0.05);胃肠安+替加氟组的实体瘤质量明显低于胃肠安组和替加氟组,差异有统计学意义(P0.05)。结论胃肠安联合替加氟对结肠癌HT29细胞及其干细胞的增殖有抑制作用。     

槐耳颗粒联合DC-CIK细胞对乳腺癌MDA-MB-231干细胞体内外杀伤作用研究

目的 观察槐耳颗粒联合树突细胞(DC)-细胞因子诱导的杀伤细胞(CIK)对乳腺癌MDA-MB-231干细胞体内外的杀伤效果.方法 用槐耳颗粒(500 mg/L)、槐耳颗粒(1000 mg/L)与乳腺癌MDA-MB-231细胞作用后,荧光倒置显微镜观察MDA-MB-231细胞形态的变化,MTT比色法检测槐耳颗粒对MDA-...     

甘草次酸修饰马钱子碱纳米给药系统的体内外肝靶向性评价

目的 制备甘草次酸（GA）修饰的马钱子碱（B）-聚乙二醇-二硫代二丙酸-单硬脂酸甘油酯（PSG）纳米粒（NPs）（B-GPSG-NPs）并评价其体内外肝靶向性。方法 采用溶剂乳化超声法制备B-GPSG-NPs和B-PSG-NPs,于透射电镜下观察其外观形态,测定其粒径、多分散性指数（PDI）、Zeta电位、包封率、载药量等理化性质。建立检测心、肝、脾、肺、肾、脑组织中马钱子碱含量的高效液相色谱法。将雌雄各半的小鼠90只随机分为3组：马钱子碱组、B-PSG-NPs组、B-GPSG-NPs组,禁食不禁水12h后,尾iv相应溶液（以马钱子碱计10mg·kg^-1）,分别于给药后10、30、60、120、180min取各组织进行HPLC检测,计算相对摄取率（Re）和靶向效率（Te）,以评价给药系统的体内靶向性。制备载异硫氰基荧光素（FITC）的FITC-B-PSG-NPs、FITC-B-GPSG-NPs,FITC、空白PSG载体制成的纳米粒（PSG-NPs）、空白GPSG载体制成的纳米粒（GPSG-NPs）以及含马钱子碱质量浓度分别为500、250、125μg·mL^-1的FITC-B-PSG-NPs和FITC-B-GPSG-NPs与CBRH-7919肝癌细胞共培养24h,荧光显微镜下观察CBRH-7919细胞对各受试物的摄取情况,以评价给药系统的体外靶向性。结果 B-GPSG-NPs的粒径为（98.91±3.62）nm,呈正态分布;PDI值为（0.221±0.006）,Zeta电位为-（19.63±0.40）mV,包封率为（78.37±1.83）%,载药量为（2.86±0.05）%;B-PSG-NPs与B-GPSG-NPs组肝脏的Re分别为1.49和1.72,明显高于其他组织;马钱子碱组肾脏Te最高,脑Te最低,而B-PSG-NPs和B-GPSG-NPs组肝脏中马钱子碱的Te明显高于其他各组织;CBRH-7919细胞摄取B-GPSG-NPs效率明显高于B-PSG-NPs,并表现出剂量相关性。结论 制备的BGPSG-NPs在体内外均表现出良好的肝靶向效应,且优于无GA修饰的B-PSG-NPs。     

湿毒清胶囊中阿魏酸的含量测定

目的建立湿毒清胶囊中阿魏酸的含量测定方法。方法采用HPLC法,uBondapak C18(4.0mm×250mm)柱,甲醇-25%冰醋酸溶液(30∶70)为流动相,检测波长323nm。结果阿魏酸在50~100μg·mL-1浓度范围内呈良好的线性关系,r=0.9999(n=5)。平均回收率100.15%,RSD为1.60%。结论该法简便、灵敏、准确,可用于湿毒清胶囊的质量控制。     

Metabolism of ferulic acid in rats

Ferulic acid is the major active constituent in many natural Chinese medicinal herbs. The metabolism of ferulic acid has been investigated using solid-phase extraction and HPLC-DAD methods that were established to separate and analyze the metabolites in urine, feces and bile. Three metabolites, identified by enzymatic hydrolysis, HPLC-DAD, HPLC-MS and MS/MS, are all glucuronic acid conjugates of ferulic acid. Ferulic acid conjugated with one glucuronic acid at different positions produces M1 and M3. Ferulic acid conjugated with two glucuronic acids produces M2, which is the main metabolite. A metabolic pathway is proposed.     

美法仑-甘草次酸复合物的合成及其体外抗肿瘤活性研究

目的设计合成美法仑–甘草次酸复合物,并对其体内外抗肿瘤活性进行研究。方法以美法仑和18α-甘草次酸为原料,通过酯化、氧化、酰化和缩合反应制备目标化合物3a和3b,结构经元素分析、MS、1H-NMR确证,并采用MTT法对其体外抗肿瘤活性进行研究,同时考察了其对正常大鼠肝细胞BRL和小鼠成纤维细胞L929的细胞毒性。结果目标化合物3a、3b的体外抗肿瘤活性明显优于母体药物18α-甘草次酸和美法仑,且对正常细胞的毒性小于氮芥类药物美法仑。结论美法仑–甘草次酸复合物3a和3b抗肿瘤活性良好,具有开发成抗肿瘤候选药物的前景。     

丙戊酸钠及其代谢产物的体外肝毒性研究

目的 研究丙戊酸钠及3个代谢产物（2-丙基-4-五烯酸、3-羟基丙戊酸、5-羟基丙戊酸）对体外人正常肝细胞L02增殖活性及对肝细胞损伤相关指标的影响。方法 实验分为对照组和实验组,对照组细胞常规培养,实验组加入丙戊酸钠及3个代谢产物,采用CCK-8法检测细胞增殖活性,PCR法检测CYP1A1、CYP1A2、PCNA、Bax及Bcl-2的mRNA相对含量,Western Blotting法检测蛋白表达,同时检测细胞上清液中谷草转氨酶（AST）、谷丙转氨酶（ALT）、乳酸脱氢酶（LDH）的含量。结果 与对照组相比,随着丙戊酸钠及3个代谢产物浓度和时间的增加,对 L02细胞增殖活性的抑制逐渐增强,CYP1A1、CYP1A2及Bax的mRNA相对含量和蛋白表达量升高,PCNA及Bcl-2的 mRNA相对含量和蛋白表达量均有下降,AST、ALT、LDH含量升高。结论 丙戊酸钠及3个代谢产物与肝毒性有关。     

通塞脉片中阿魏酸的含量测定

目的建立通塞脉片中阿魏酸的含量测定方法。方法采用HPLC法,uBondapak C18（4．6mm×250mm）,甲醇,25％冰醋酸溶液（35：65）为流动相。检测波长323nm。结果阿魏酸在0．01034—0．1034mg·mL^-1浓度范围内呈良好的线性关系,r=0．9999（n=6）。平均回收率98．68％,RSD为2．09％。结论该法简便、灵敏、准确,可用于通塞脉片的质量控制。     

HPLC法测定健脑丸中阿魏酸含量

目的:测定健脑丸中阿魏酸含量。方法:采用HPL C法,色谱柱为C18,流动相为甲醇-水-冰醋酸(38∶6 2∶0 .1) ,检测波长为32 3nm。结果:平均加样回收率为99.0 7%。结论:方法简便、快速、灵敏度高,可用于该品的质量控制     

全反式维甲酸固体脂质纳米粒的制备及体内外评价

目的以山嵛酸甘油酯(Compritol 888 ATO)为脂质材料，采用超声分散法制备维甲酸固体脂质纳米粒，并考察其体内外性质。方法选用脂溶性较高的维甲酸作为模型药物，采用超声分散法制备固体脂质纳米粒，并对其各种理化性质进行研究。考察了纳米粒的体外释放，以维甲酸溶液剂为对照，测定了两种纳米粒在大鼠体内的药代动力学参数。结果采用超声分散法可以简便、快速制备得到两种维甲酸固体脂质纳米粒，透射电镜测得纳米粒为圆球状，大小均匀。动态光散射法测得平均粒径分别为(158±9) nm和(89±11) nm。于4 ℃放置1年粒径无明显变化，载药量为3.3%，包封率大于95%。药物体外释放符合Weibull方程。与对照组相比，两种维甲酸固体脂质纳米粒静脉注射后药物在血液中的滞留时间显著延长。结论超声分散法适用于固体脂质纳米粒的制备。     

去甲泽拉木醛的体外抗真菌作用研究

目的 研究去甲泽拉木醛的体外抗真菌作用。方法 采用微量液基稀释法测定去甲泽拉木醛与氟康唑单独应用于23株真菌的最低抑菌浓度（MIC）,以棋盘式微量液基稀释法测定两药联合抗耐药白念珠菌的协同指数（FICI）,判断两药联合抗菌效果;并通过纸片扩散实验直观验证两药联合的协同作用。最后通过CCK-8法测定去甲泽拉木醛的细胞毒性。结果 去甲泽拉木醛单用时呈现广谱的抗真菌作用,MIC范围为4~32 g/L。两药联用时,可将氟康唑的有效浓度从大于64 g/L降至0.25 g/L,FICI值介于0.129~0.254之间,两药表现出协同抗耐药白念珠菌作用。CCK-8结果显示,去甲泽拉木醛在高于MIC值4倍浓度下才展示出细胞毒性。结论 去甲泽拉木醛表现出较好的抗真菌作用,与氟康唑联合时有很好的协同效果,且毒性较低。     

Assessment of the potential irritancy of oleic acid on human skin: Evaluation in vitro and in vivo

As skin barrier modulating compounds, fatty acids are frequently used in formulations for transdermal or topical delivery. In this study the effects of oleic acid on keratinocytes in vitro was compared with its in vivo skin irritancy in humans. Dose- and time-dependent effects of oleic acid were examined in submerged human keratinocyte cultures, in reconstructed human epidermis (RE-DED), and in excised human skin, using alterations in morphology and changes in interleukin-1 mRNA levels as endpoints. In vitro results were compared with responses of living human skin after topical application of oleic acid, using non-invasive bioengineering methods. Direct interaction of oleic acid and submerged keratinocyte cultures resulted in cell toxicity at very low concentrations of the fatty acid. By contrast, when oleic acid was applied topically on RE-DED or on excised skin, no alterations in morphology were observed. Modulation of stratum corneum thickness indicated a key role of the stratum corneum barrier in the control of oleic acid-induced toxicity. In agreement with these findings, no epidermal tissue damage was seen in vivo, whereas oleic acid induced a mild but clearly visible skin irritation and inflammatory cells were present in the upper dermal blood vessels. Small amounts of oleic acid induced IL-1 mRNA expression in submerged keratinocyte cultures, whereas in RE-DED and in excised skin, IL-1 mRNA levels were increased only when the concentration applied topically was at least two orders of magnitude higher. It is concluded that minute amounts of oleic acid are sufficient to cause local (i.e. inside the viable epidermis) modulation of cytokine production. These concentrations do not affect morphology but induce skin irritation in vivo. To achieve comparable effects in the skin, much higher topical doses are needed than expected according to the locally required levels, owing to the rate-limiting transport of the fatty acid across the stratum corneum barrier.     

HPLC法测定宫炎康胶囊中阿魏酸的含量

目的建立高效液相色谱法测定宫炎康胶囊中阿魏酸的含量。方法采用Aglient C18为色谱柱;流动相:乙腈-0.4%磷酸溶液(15∶85),流速为1.0 mL·min-1,检测波长为316 nm。结果阿魏酸在2.58¹2.88μg·mL-1范围内线性关系良好,回收率为98.4%,RSD=1.1%。结论方法简便,结果准确,能有效控制宫炎康胶囊的质量。