Enhancement of rhubarb extract solubility and bioactivity by 2-hydroxypropyl-β-cyclodextrin

Rhubarb is a traditional Chinese medicinal herb, and the ethanolic extract of rhubarb consists of active anthraquinones, which are hydrophobic and have antiproliferative effects on hepatoma cell lines. To increase the aqueous solubility of rhubarb and study the consequent bioavailability, the ethanolic extract of rhubarb was complexed with 2-hydroxypropyl-β-cyclodextrin (HP-β-CD), a cyclic oligosaccharide that has a hydrophilic outer surface and a hydrophobic central cavity, to form a rhubarb–HP-β-CD complex. This complex was characterized by performing nuclear magnetic resonance spectroscopy, two-dimensional rotating frame spectroscopy and thin layer chromatography to confirm the inclusion of anthraquinones from rhubarb extract in HP-β-CD (weight ratio of rhubarb extract:HP-β-CD = 1:9). We investigated the effects of complexing rhubarb extract with HP-β-CD on the growth of Huh7 and HepG2 cells by performing cytotoxicity analysis, cellular uptake test, and colony formation assay. Our results showed that complexation of rhubarb extract with HP-β-CD increased the aqueous solubility and bioavailability of rhubarb and thus enhanced its effect on hepatoma cells.     

火木层孔菌发酵菌粉及其组分对小鼠移植性肝癌H22的体内生长作用

采用肝癌H22荷瘤小鼠的肿瘤模型,对火木层孔菌(桑黄)Phellinus igniarius发酵菌粉及其各提取物组分的体内抗肿瘤活性进行评价。结果表明,火木层孔菌发酵菌粉及其各提取物组分对荷瘤小鼠肝癌H22都具有一定的抗肿瘤作用并能延长荷瘤小鼠生存期,其中,火木层孔菌发酵菌粉(1,000mg/kg/d)及其组分I(I多糖组分)(360mg/kg/d)具有较为显著的抗肿瘤作用,抑瘤率分别为33.5%和40.3%。组织病理学研究结果表明在菌粉及其多糖组分作用后,肿瘤细胞坏死细胞明显增多,免疫组化检测表明火木层孔菌发酵菌粉及其多糖组分能明显的降低瘤组织中Bcl-2基因蛋白的表达,提高小鼠瘤组织中的Bax基因蛋白的表达。火木层孔菌发酵菌粉及其多糖组分具有较好的体内抗肿瘤活性。     

细胞松弛素D、秋水仙素作用下大鼠肝癌细胞中黏弹性的研究

采用微管吮吸技术测定大鼠肝癌细胞的黏弹性;研究了秋水仙素、细胞松弛素D以及两者混合作用后对于肝癌细胞黏弹性的影响。结果表明:用CD处理癌细胞后发现癌细胞的弹性系数K1明显下降。与对照组相比:微丝骨架被CD抑制后,在加入Col后肝癌细胞的弹性系数K1显著降低;而微管骨架被col抑制后,在加入CD后肝癌细胞的弹性系数K1、K2和μ无明显变化。提示在微管骨架系统完整的情况下,微丝对肝癌细胞的黏弹性系数的的影响起主要作用。而微管骨架系统受到破坏后,微丝需借助于微管网络的作用来影响细胞的黏弹性。本研究对揭示癌细胞中骨架系统之间的交互作用对于细胞黏弹性的影响提高实验依据。     

丹参酮通过VEGF/VEGFR信号通路抑制肝癌细胞迁移和侵袭能力的实验研究

目的:探究丹参酮通过VEGF/VEGFR信号通路抑制肝癌细胞迁移和侵袭能力的机制。方法:体外培养人肝癌细胞Hep3B、HepG2,并分为:实验组、阳性对照组和空白对照组,实验组使用丹参酮处理,阳性对照组使用阿霉素处理,空白对照为加入10μL DMSO或生理盐水,使用CCK8检测肝癌细胞的增殖情况;分别加入浓度为31μmol/L和2.5μmol/L的丹参酮及阿霉素显微镜下观察细胞形态变化;使用流式细胞术检测肝癌细胞的凋亡情况和细胞周期情况;使用Western blot检测肝癌细胞VEGF及VEGFR蛋白表达情况。结果:MTT实验结果显示,随着丹参酮和阿霉素使用浓度的升高人肝癌细胞Hep3B、HepG2生长受到显著的抑制(P0.05);显微镜下观察发现丹参酮可以抑制肝癌肿瘤细胞增殖;流式细胞检测发现相比空白对照组,阳性对照组和实验组(人肝癌细胞Hep3B、HepG2)细胞凋亡率显著增加(P0.05);相比空白对照组,实验组和阳性对照组G0/G1期细胞百分比显著增加(P0.05),S期和G2/M期细胞百分比显著降低(P0.05);蛋白质印记实验结果显示,相比空白对照组,实验组和阳性对照组VEGF及VEGFR蛋白表达显著降低(P0.05),且实验组表达显著低于阳性对照组(P0.05)。结论:参酮通可以通过抑制VEGF/VEGFR信号通路,将肝癌细胞分裂阻滞在G0/G1,达到抑制肝癌细胞的增殖及迁移和侵袭能力的效果。     

Ultrasensitive detection of ferritin in human serum by Western blotting based on quantum dots-labeled avidin-biotin system

Various methods have been applied for serum ferritin detection, however, these methods still have some limitations. Over the last few years, quantum dots (QDs) have become very attractive for immunoassays because of their enormous potentials in ultrasensitive analysis. In this study, a Western blotting method combined with QDs‐labeled avidin–biotin system for detecting human serum ferritin was described. Meanwhile, the traditional diaminobenzidine (DAB)–horseradish peroxidase (HRP) method had been compared with the method. The linearity of this QDs‐based Western blotting method was from 0.27 to 1.1 ng, and the quantification limit was 0.27 ng, the sensitivity was up to pictogram values. Real serum samples such as hepatoma, thalassemia patient and normal individual sera were analyzed, the analysis results demonstrated that there was significant difference in the concentrations of ferritin between patients and normal individual serum. Furthermore, the recovery of ferritin from the serum samples of patients ranged from 98.15 to 119.67%, and the RSD (relative standard deviation) ranged from 8.73 to 11.61%, the repeatabilities were well within the acceptable range, which revealed that this method is a stable and reproducible method for detecting serum ferritin and have potential application prospect in clinical laboratory of serum ferritin detection.     

海藻色素糖蛋白对肝癌细胞Caspase-3和Bax蛋白表达的影响

目的：研究麒麟菜海藻色素糖蛋白（SPG）对肝癌细胞Caspase-3和Bax蛋白表达的影响。方法：将50只皮下接种H22肝癌细胞株的小鼠随机分为5组,每组10只。高、中、低剂量组分别每天经口灌胃给予100、50、10mg/kg的SPG,肿瘤对照组灌胃生理盐水,连续10d。环磷酰胺组隔天腹腔注射环磷酰胺20mg/kg.bw。取肝癌组织用MTT法测定各组肝癌细胞增殖活性,免疫组化法检测各组肝癌组织Caspase-3和Bax蛋白表达水平。结果：高剂量组Caspase-3和Bax蛋白表达率分别为40.20%和38.10%,而肿瘤对照组分别为5.00%和4.68%,差异均有显著性（P〈0.05）。高剂量SPG组和肿瘤对照组的肝癌细胞增殖活性分别为0.711±0.028和1.135±0.032,差别有显著性（P〈0.05）。结论：SPG可促进肝癌细胞Caspase-3和Bax蛋白表达,诱发肝癌细胞凋亡。     

RMP基因干扰对肝癌细胞周期的影响

目的:建立RMP(RPB5-Mediating Protein)基因干扰的稳定细胞株,研究RMP基因干扰对正常肝细胞及其肝癌细胞周期的影响.方法:构建RMP基因的短发卡RNA(short-hairpin RNA,shRNA)真核表达载体pGPU6-Neo-RMPi-484.通过脂质体转染的方法转染到SMMC-7721肝...     

Silkworm powder containing manganese superoxide dismutase regulated the immunity and inhibited the growth of Hepatoma 22 cell in mice

The effects of SOD contained silkworm powder on immune regulation and inhibition against Hepatoma 22 tumor cells in vivo were investigated. The activity of natural killer cell (NK) and the ConA-stimulated spleen proliferation were measured. The results found that the SOD-contained silkworm powder caused an enhancement on NK cell activity, which implied this material modulated the immune system in mice in vivo. The NK cell activities of Hepatoma 22 tumor modeled mice treated with silkworm powder including SOD were increased significantly compared to a modeled control and silkworm powder without SOD, reaching 36.18%. In addition, the ConA-stimulated spleen proliferation of SOD treated mice was higher than that of the controls. The treatment of SOD contained silkworm powder presented 40.3% of average inhibition rate to Hepatoma 22 tumor, showing stronger inhibition against tumor. There were no significant difference in body weight between modeled control and SOD silkworm powder feeding in Hepatoma 22 tumor modeled mice, suggesting the SOD silkworm powder is safety as an inhibitant to tumor. In conclusion, these findings demonstrate that administration of silkworm powder containing SOD results in activation of NK cells and immunity, suggesting the silkworm powder containing SOD plays a positive role in tumor inhibition.     

新型磁性纳米脂质复合物对肝癌细胞和肝细胞的作用研究

目的:研究新型磁性纳米脂质复合物对肝癌细胞和肝细胞的作用。方法:将肝癌细胞(Hep-G2)和肝细胞(L-02)根据加入的不同浓度新型纳米脂质复合物,各自分为空白对照组、不同浓度含铁脂质体组、空脂质体组,用MTT法观察各组细胞毒性,筛选出适合的浓度和时间;用普鲁士蓝染色、电镜检测观察各组细胞吞噬情况;用原子分光光度计观察各组细胞内铁含量;用细胞磁共振观察新型纳米脂质复合物体外显影效果。结果:新型磁性纳米脂质复合物含铁浓度在0.238μg/ml以上时,24小时有较强的细胞毒性,对肝癌细胞和肝细胞的抑制率均超过40%,故以下实验选择含铁浓度在0.238μg/ml以下;肝癌细胞和肝细胞与各浓度新型磁性纳米脂质复合物共同培养,24小时普鲁士蓝染色最高阳性率分别为5.5%和1.25%,24小时细胞内铁含量最高值分别为0.675pg/cell和0.460pg/cell;24小时电镜观察肝癌细胞和肝细胞空白对照组均未见颗粒样物质,各含铁脂质体组和空脂质体组均可见颗粒样物质;24小时肝癌细胞和肝细胞磁共振感兴趣区均未见明显信号表达。结论:新型磁性纳米脂质复合物含铁浓度在0.238μg/ml以下时,24小时细胞毒性较小;肝癌细胞对新型磁性纳米脂质复合物的吞噬作用稍高于肝细胞对其的吞噬;进入细胞内的新型磁性纳米脂质复合物含铁量低,体外显影效果不佳。     

5- 脱氧杂氮胞苷对人肝癌细胞HepG2 增殖及beclin1 表达的影响

目的:观察5-脱氧杂氮胞苷(5-aza-CdR)对HepG2细胞生长抑制及beclin1表达的影响,以探讨其抗肿瘤发生的潜在机制。方法:采用四甲基偶氮唑盐(MTT)法检测5-aza-CdR对HepG2细胞的生长抑制;用相差显微镜观察不同药物浓度下不同时间段的肝癌细胞形态学改变;采用RT-PCR法和Western blot法检测5-Aza-CdR对抑癌基因beclin1的mRNA和蛋白表达的影响。结果:5-aza-CdR可抑制HepG2细胞生长,呈剂量依赖性,并上调beclin1的mRNA和蛋白的表达。结果:显示102.4umol/L5-aza-C-dR作用72小时细胞增殖抑制率最高,可达(84.3±3.31)%,beclin1的mRNA和蛋白表达上调最明显,与对照组相比差异有统计学意义。结论:5-aza-CdR可抑制HepG2细胞增殖,其机制可能是通过恢复某些抑癌基因的表达,上调beclin1的mRNA和蛋白表达。