合欢皮提取物抑制血管生成的体内药效学研究

为了探讨合欢皮提取物体内对血管生成的抑制作用。本文采用MTT法和Matrigel plug方法考察合欢皮提取物对人血管内皮细胞(HMEC)增殖和体内Matrigel胶小管形成的影响,运用Lewis肺癌转移小鼠模型,观察合欢皮提取物对小鼠Lewis肺癌的疗效。结果表明合欢皮提取物可明显抑制HMEC细胞的体外增殖(IC50为30μg/mL),并且呈明显的剂量依赖性,同时具有抑制Matrigel plug中的血管新生的作用。合欢皮提取物还能够明显抑制Lewis肺癌肿瘤细胞生长,减少转移病灶。提示合欢皮提取物具有明显抑制血管生成的作用,有可能成为有效的血管生成抑制剂。     

灵芝多糖抗肿瘤作用的免疫分子机制研究

灵芝多糖是一种从灵芝实体中被提取出来的多糖组份,灵芝多糖极易溶于水。很多科学研究表明,灵芝多糖具有一定的免疫调节功能以及一定的抗肿瘤等作用,并且认为在体外灵芝多糖并没有直接的对肿瘤细胞有抑制作用或者杀伤作用,反而在体内灵芝多糖则具有很好的抗肿瘤作用。所以,通常情况下,对灵芝多糖的抗肿瘤作用的研究主要就是通过增加免疫功能,尽管我国很多科学家已经从免疫功能的角度对灵芝多糖的抗肿瘤作用进行研究,但确切的机制并没有明确。因此文章结合具体实验对灵芝多糖的抗肿瘤作用的免疫分子机制进行深入研究。     

灵芝菌丝体中一个三萜类化合物的核磁归属及活性初探

通过液体振荡-静置两阶段发酵获得灵芝菌丝体,并采用硅胶柱色谱层析、反相柱层析和甲醇重结晶的方法,从中分离得到4个三萜类化合物。根据NMR、MS等波谱数据分析,化合物分别被鉴定为lanosta-7,9(11),24-trien-3α-acetoxy-26-oic acid（1）、灵芝酸R（2）、灵芝酸T（3）和灵芝酸S（4）,其中化合物1的核磁信号全归属为首次报道。4个三萜类化合物均具有较好的抑制肿瘤细胞L1210及K562增殖的活性,且化合物1的体外抗肿瘤活性为首次证实,其对肿瘤细胞L1210及K562增殖的半数抑制浓度IC₅₀分别为22.17μmol/L和54.79μmol/L。     

一种无孢灵芝菌丝体的活性成分

利用制备型高效液相、凝胶层析、制备型薄层色谱等方法对无孢灵芝龙芝2号的固体发酵菌丝体进行分离纯化,通过波谱分析,化合物分别鉴定为灵芝酸P（1）,灵芝酸T1（2）,灵芝酸Mk（3）,灵芝酸S（4）,灵芝酸T（5）,ganodermanondiol（6）,灵芝酸Me（7）,5α, 8α-epidioxyergosta-6, 22-dien-3β-ol（8）,灵芝酸R（9）,lanosta-7, 9(11), 24-trien-3α-hydroxy-26-oic acid（10）,ganodermenonol（11）。其中灵芝酸T1为首次发现的天然产物。体外细胞实验证实,11种化合物对肿瘤细胞L1210的增殖均有很强的抑制作用,其增殖抑制的IC₅₀值均在39.69μmol/L以下。     

层迭灵芝子实体的体外抗肿瘤及免疫活性

【背景】层迭灵芝Ganoderma lobatum是灵芝属中的一个种,在民间有药用历史,但缺乏对其化学成分和药理活性的科学研究。【目的】以赤芝Ganoderma lingzhi子实体为参照,研究对比层迭灵芝子实体的抗肿瘤及免疫活性的强弱,探讨层迭灵芝的药用价值。【方法】采用化学分析及仪器分析的方法,比较2种灵芝子实体中三萜及多糖含量差异,并进行体外抗肿瘤及免疫活性研究。【结果】层迭灵芝和赤芝的子实体中三萜含量差异不大,分别为1.14%和1.21%,但2种灵芝中三萜化合物的种类差异较大。层迭灵芝子实体中的多糖含量较赤芝稍高,分别为3.60%和2.67%,2种子实体中多糖的重均分子量分布特征有所差别。2种灵芝醇提物对肿瘤细胞K562及SW620的增殖均具有一定的抑制活性,其中,层迭灵芝对SW620细胞具有较强的抑制活性,其IC50值达到了52.5μg/mL。2种灵芝水提物可以促进RAW 264.7细胞释放NO,说明两者均具有一定的免疫活性。【结论】层迭灵芝具有较好的抗肿瘤及免疫活性,可以作为药用开发的原料来源。     

纳米级灵芝子实体粉末和破壁灵芝孢子粉体外抗肿瘤活性研究

对纳米级灵芝子实体粉末及破壁灵芝孢子粉石油醚提取物(PE)、氯仿提取物(CE)、丙酮(AE)、甲醇提取物(ME)、水提取物(WE)与灵芝子实体及灵芝孢子提取量进行对比,利用GC-MS联用仪对石油醚提取物进行了成分分析鉴定,对水提物中总糖进行了含量测定,并利用宫颈癌细胞Hela和晶体上皮细胞SRA01/04进行了体外增殖作用和剂量效应关系研究,为灵芝资源的保护及进一步开发利用提供理论基础。结果表明,纳米化灵芝子实体及破壁灵芝孢子不同溶剂提取量显著增加,纳米级灵芝子实体粉末水提取物具有抑制宫颈癌细胞Hela和晶体上皮细胞SRA01/04增殖的作用。破壁灵芝孢子各溶剂提取物对宫颈癌细胞Hela和晶体上皮细胞SRA01/04没有明显的增殖抑制作用。     

灵芝子实体中三萜类物质抗肿瘤活性的谱效关系

通过HPLC指纹图谱结合多元线性回归分析对不同产地灵芝子实体的功效性特征进行评价,为寻找灵芝中活性三萜提供理论依据。利用高效液相分析方法,结合样品对肿瘤细胞L1210的增殖抑制率,运用“中药色谱指纹图谱相似度评价系统2004A版”软件和多元线性回归分析11批不同品种灵芝子实体中的三萜活性成分。样品与标准指纹图谱的相似度均在0.9以上,共标定了12个共有物质峰,其中与抗L1210肿瘤细胞活性关系密切的三萜物质有灵芝酸C₂、灵芝酸G、灵芝烯酸B、灵芝烯酸A、灵芝酸K、灵芝酸A、灵芝酸F和灵芝醛A。     

糖肽对小鼠Lewis肺癌实验性转移作用的初步病理观察

我们曾报道从小鼠Lewis肺癌组织通过蛋白水解酶及分子筛层析分离的总糖肽,在体外可明显地抑制某些肿瘤细胞及分离的层粘连蛋受体与基膜成分层粘连蛋白的识别和结合。本文报告将此糖肽与Lewis肺癌细胞混合,通过尾静脉注入小鼠体内,对实验性癌转移的抑制作用。初步病理结果表明,此糖肽几乎可以完全抑制实验性转移瘤的形成,保护小鼠不死于癌转移。提示糖肽可能具有阻断癌转移之作用。将实验组一部分存活小鼠再行同种癌细胞皮下接种,可以照常成瘤。表明糖肽阻抑实验性癌转移的效能可能并非调动了宿主的免疫机制所致。糖肽还可减慢皮下接种的癌细胞的生长速度,但对癌细胞并无直接毒性作用。     

栽培条件对灵芝子实体的三萜组成及其活性的影响

以不同灵芝品种、不同栽培基质、不同管理方式、不同生长时期获得的灵芝子实体为原料,用95%乙醇超声提取,对提取物进行了含量测定、三萜组成分析和体外抗肿瘤实验。结果表明,不同子实体醇提物中三萜和甾醇类物质的含量在4.60-6.20mg/g之间;高效液相分析发现10种三萜化合物的种类和含量在样品间存在明显差异。所有灵芝子实体醇提物对肿瘤细胞L1210的增殖均有一定的抑制作用,开伞期的灵芝子实体醇提物抑制肿瘤增殖的活性优于其他生长时期。     

灵芝多糖抗肿瘤作用的初步探讨

目的 :初步探讨灵芝多糖对小鼠肿瘤的抑制作用。方法 :观察灵芝多糖对 S180 、U14腹水型荷瘤小鼠生存期的影响 ,了解灵芝多糖对 S180 、U14、H2 2 肿瘤细胞有无直接杀伤作用 ,以寻找治疗肿瘤的新方法。结果 :灵芝多糖能够显著延长荷瘤小鼠的生存期 (P<0 .0 5) ,治疗组小鼠的生存期比对照组小鼠的生存期明显延长 ,但是灵芝多糖对肿瘤细胞无直接杀伤作用 (P<0 .0 5)。结论 :灵芝多糖抗肿瘤作用是通过机体的免疫系统介导的。     

一些抗癌药物对荷瘤小鼠血清中唾液酸含量的影响

测定荷六种小鼠肿瘤S180肉瘤(实体型和腹水型),腹水肝癌(HepA),艾氏腹水瘤(EC),白血病P388和Lewis肺癌的小鼠腹水和血清中唾液酸含量,结果显示血清中唾液酸含量与肿瘤生长、肿瘤类型有关。腹水中唾液酸含量高,推测肿瘤能比正常组织产生更多唾液酸。对四种腹水肿瘤用阴离子交换树脂层析鉴定,发现HepA腹水中葡萄糖代唾液酸(NcuGc)含量明显低于其它三种腹水瘤。还研究了十几种抗癌药物对荷S180和Lewis肺癌小鼠血清中唾液酸含量的影响。发现吗丙嗪(probimane)和顺铂(DDP)能降低荷瘤小鼠血清中唾液酸含量,提示此二药物在肿瘤治疗中更具选择性。     

Vaccination with TCL plus MHSP65 induces anti-lung cancer immunity in mice

To develop effective anti-lung cancer vaccines, we directly mixed mycobacterial heat shock protein 65 (MHSP65) and tumor cell lysate (TCL) from Lewis lung cancer cells in vitro and tested its efficacy on stimulating anti-tumor immunity. Our results showed that MHSP65–TCL immunization significantly inhibited the growth of lung cancer in mice and prolonged the survival of lung cancer bearing mice. In vivo and in vitro data suggest that MHSP65–TCL could induce specific CTL responses and non-specific immunity, both of which could contribute to the tumor inhibition. Thus, this report provides an easy approach to prepare an efficient TCL based tumor vaccine.     

IRM-2小鼠移植性肿瘤模型的生物学特性

目的观察IRM-2小鼠和C57BL/6小鼠接种Lewis肺癌生物学特性的对比研究。方法取肿瘤组织研磨,用生理盐水稀释成2×10^6/mL,取细胞悬液接种于IRM-2小鼠和C57BL/6小鼠腋下,0.2 mL/只。观察两品系肿瘤生长、荷瘤鼠生存时间,外周血细胞及病理指标变化。结果两品系小鼠成瘤率均是100%,荷瘤鼠存活时间无明显差异,IRM-2小鼠荷瘤鼠体重净增长明显高于C57BL/6荷瘤小鼠（P〈0.05）。白细胞分类及病理指标变化无明显差别。结论IRM-2小鼠与C57BL/6小鼠Lewis肺癌模型生物学特性基本一致,IRM-2小鼠可以建立稳定的Lewis肺癌肿瘤模型应用于实验研究。     

Asian ginseng extract inhibits in vitro and in vivo growth of mouse lewis lung carcinoma via modulation of ERK‐p53 and NF‐κB signaling

Asian ginseng (AG) is the most commonly used medicinal herb in Asian countries. It is often prescribed for cancer patients as a complementary remedy. However, whether AG in fact benefits cancer patients remains unknown because some studies reported that AG facilitates tumor growth, which contradicts its usage as a dietary remedy to cancer patients. In addition, most of research works on ginseng for anti‐cancer were using single ginsenoside rather than whole root extracts used in clinics. Thus, intensive studies using the type of ginseng as its clinical form are necessary to validate its benefits to cancer patients. In this study, anti‐tumor potency and underlying molecular mechanisms of the ethanol extract of AG (EAG) were examined in mice with Lewis lung carcinoma (LLC‐1). We showed that EAG significantly suppressed tumor growth in LLC‐1‐bearing mice with concomitant down‐regulation of PCNA proliferative marker, and it exhibited specific cytotoxicity to cancer cells. EAG also induced MAPK and p53 signaling in LLC‐1 cells, which suppressed cyclin B–cdc2 complex and in turn induced G2–M arrest and apoptosis. Although EAG could activate NF‐κB signaling, the proteasome inhibitor of MG‐132 could effectively prevent NF‐κB targeted gene expression induced by EAG and then sensitize LLC‐1 cells to induce EAG‐mediated apoptosis. Collectively, EAG in a relatively high dose significantly suppressed tumor growth in LLC‐1‐bearing mice, indicating that AG may benefit lung cancer patients as a dietary supplement. This is the first report demonstrating possible combination of EAG with proteasome inhibitors could be a novel strategy in anti‐cancer treatment. J. Cell. Biochem. 111: 899–910, 2010. © 2010 Wiley‐Liss, Inc.     

小鼠Lewis肺癌组织中糖肽的分离纯化及其对肿瘤细胞与层粘连蛋白基质粘着的影响

小鼠Lewis肺癌组织经氯仿甲醇去除脂类,用木瓜蛋白酶消化,再经Sephadex柱层析分离得到总糖肽。它有明显地抑制小鼠Lewis肺癌细胞,S180肉瘤细胞及人巨细胞肺癌细胞与层粘连蛋白(Laminin,LN)基质粘着的作用;对层粘连蛋白受体(LN-R)与其配体的识别及结合也具有同样明显的阻断效应。糖肽的上述作用均具有剂量依赖性。进一步经ConA-Sepharose CL-4B亲和层析将总糖肽分为三个部分。与ConA不结合的糖肽部分对Lewis肺癌细胞与LN基质的粘着也具有剂量性抑制作用。     

IL-10 promotes resistance to apoptosis and metastatic potential in lung tumor cell lines

Treatment of non-small cell lung carcinoma (NSCLC) remains at a disappointingly low success rate. Not only is metastatic spread common in NSCLC, but therapeutic success decreases dramatically once metastases are present. Understanding factors which contribute to poor prognosis in NSCLC is critical for development of more successful therapeutic approaches. Interleukin-10 (IL-10) expression has been shown in several studies to correlate with a poorer prognosis in NSCLC; however, the mechanisms by which IL-10 affects lung tumor growth and metastases are unclear. The goal of this study was to evaluate the effects of tumor-derived IL-10 on the growth and metastasis of lung cancer cells in a murine model. Lewis lung carcinoma cells were stably transfected with the chicken ovalbumin gene (cOVA) as a model tumor antigen (LL43 tumor cells) and subsequently transfected with the murine IL-10 gene (LL43-10 tumor cells). Subcutaneous growth of the LL43 tumor cells was not affected by expression of IL-10. However, LL43-10 tumors had a fourfold increase in tumor microvessel density, as indicated by CD31 staining. Metastatic potential was also increased in IL-10-expressing lung tumor cells, leading to a greater number of tumor cells in lymph nodes draining the primary tumor site. Finally, exposure of Lewis lung tumor cells in vitro to exogenous IL-10 dramatically increased their resistance to UV-induced apoptosis. These results indicate that a primary effect of IL-10 on lung cancer cells may be to increase their metastatic potential by promoting angiogenesis and resistance to apoptosis.     

桦木酸对H22荷瘤小鼠肿瘤细胞凋亡及细胞周期的影响

研究桦木酸对H22荷瘤小鼠生命延长率、肿瘤细胞凋亡及细胞周期的影响。结果表明桦木酸能够明显延长H22荷瘤小鼠生生存时间,其中低、中剂量效果显著（P〈0．05）;利用DNA结合性荧光探针直接对细胞DNA染色后FCM分析,桦木酸可能是通过影响H22肿瘤细胞S期而诱导肿瘤细胞凋亡。     

CDA-2, a Urinary Preparation,Inhibits Lung Cancer Development through the Suppression of NF-kappaB Activation in Myeloid Cell

CDA-2 (cell differentiation agent 2), a urinary preparation, has potent anti- proliferative and pro-apoptotic properties in cancer cells. However, the mechanisms of tumor inhibitory action of CDA-2 are far from clear, and especially there was no report on lung cancer. Here we demonstrate that CDA-2 and its main component phenylacetylglutamine (PG) reduce the metastatic lung tumor growth, and increases survival time after inoculation with Lewis lung carcinoma (LLC) cells in a dose-dependent manner in C57BL6 mice. Proliferative program analysis in cancer cells revealed a fundamental impact of CDA-2 and PG on proliferation and apoptosis, including Bcl-2, Bcl-XL, cIAP1, Survivin, PCNA, Ki-67 proteins and TUNEL assays. CDA-2 and PG significantly reduced NF-κB DNA-binding activity in lung cancer cells and in alveolar macrophages of tumor bearing mice and especially decreased the release of inflammatory factors including TNFα, IL-6, and KC. Furthermore, CDA-2 and PG decrease the expressions of TLR2, TLR6, and CD14, but not TLR1, TLR3, TLR4, and TLR9 in bone-marrow-derived macrophages (BMDM) of mice stimulated by LLC-conditioned medium (LLC-CM). Over-expressing TLR2 in BMDM prevented CDA-2 and PG from inhibiting NF-κB activation, as well as induction of TNFα and IL-6. TLR2:TLR6 complexes mediate the effect of NF-κB inactivation by CDA-2. In conclusion, CDA-2 potently inhibits lung tumor development by reduction of the inflammation in lung through suppression of NF-κB activation in myeloid cells, associating with modulation of TLR2 signaling.     

藻红蛋白亚基光敏剂对小鼠移植瘤作用的超微结构研究

目的：从形态学角度探讨藻红蛋白(R-PE)β亚基光动力学抗肿瘤效果及其作用机理。方法：用不同密度的波长为496nm的氩离子激光对S180小鼠移植瘤进行β亚基光动力学治疗,并对治疗后的瘤体进行透射电镜的形态学观察。结果：用100μg／m1的β亚基,在200J/cm2激光照射剂量条件下治愈了瘤体直径为0．5cm-0．7cm大小的小鼠移植瘤,发现瘤组织中引起细胞死亡的途经有差异,被PDT抑制的肿瘤内部细胞表现出典型的凋亡细胞特征。结论：R-PE β亚基具较强的光动力学抗肿瘤效果,光动力治疗机理可能涉及肿瘤内部细胞死亡主要是凋亡途径而瘤周为坏死,且与血管系统破坏及白细胞参与的抗炎症反应相关。