雷公藤甲素通过抗ER磷酸化抑制小鼠4T1乳腺癌增殖作用研究

雷公藤甲素（triptolide,TP）具有显著的抑制肿瘤作用。本研究采用MTT法和结晶紫染色法检测TP对小鼠乳腺癌细胞4T1增殖的影响;流式细胞仪检测TP诱导的4T1细胞凋亡率;Western blot检测4T1中相关蛋白雌激素受体α（estrogen reporter α,ERα）,磷酸化ERα（p-ERα）,ERβ,p-ERβ,细胞外信号调节的蛋白激酶（ERK）,p-ERK,p38,p-p38,c-Jun氨基末端激酶（SAPK/JNK）,p-SAPK/JNK的表达。给药30 d后,活体荧光示踪技术观察TP对4T1-Luc细胞接种的BALB/c 雌性小鼠乳腺癌的荧光强度,同时检测肿瘤体积和质量,计算抑瘤率。肿瘤组织苏木素-伊红（HE）染色后观察病理变化;免疫组织化学法（IHC）分析肿瘤组织ERα及ERβ的表达情况。发现TP体外浓度为100,10,1,0.1 μmol·L^-1时可显著抑制4T1乳腺癌细胞的增殖,诱导细胞凋亡。TP剂量为200 μg·kg^-1时能抑制小鼠乳腺癌的增殖,TP的抗乳腺癌增殖作用与降低ERα,ERβ的磷酸化密切相关,但与MAPK通路的活化无明显相关性。     

雷公藤内脂醇对正常人和SLE患者PBMCs中细胞增殖的影响

目的:研究雷公藤内酯醇(TPL)对正常人和系统性红斑狼疮(SLE)患者外周血单核细胞(PBMCs)增殖的影响,以探讨雷公藤内酯醇在SLE治疗中的意义。方法:采用MTT法检测雷公藤内酯醇对正常人和SLE患者PBMC细胞的抑制率。结果:TPL对正常人和SLE患者PBMCs均有明显的生长抑制作用,并呈浓度和时间依赖;与正常人相比较,TPL对于SLE患者的抑制作用更明显。结论:根据TPL作用于正常人和SLE患者PBMCs的半数抑制浓度差别说明可以找一个合适的剂量对SLE发挥最大治疗作用,而对正常人无明显影响。     

Mucuna pruriens proves more effective than L-DOPA in Parkinson's disease animal model

Seeds of the Mucuna pruriens plant, now known to contain L -DOPA, have long been used for the treatment of Parkinson's disease patients in ancient Eastern Indian ethnotherapeutics. Following validation of the intrastraital 6-OHDA injection with amphetamine in the parkinsonian rat model, the animals were fed synthetic L -DOPA (125 or 250 mg/kg) or Mucuna pruriens endocarp (MPE, 2.5 or 5.0 g/kg) mixed with rat chow (n =6, for each dose and drug). Controls received no drug. An additional dose of L -DOPA or MPE in the same doses plus carbidopa (50 mg/kg) were administered via gavage (controls received only carbidopa 50 mg/kg) 1 h prior to testing with rotometer. Contralateral rotation (to the side of the 6-OHDA lesion) (CLR) was recorded for 240 min as a measure of antiparkinsonian activity. Results indicated that dose for dose, MPE showed twice the antiparkinsonian activity compared with synthetic L -DOPA in inducing CLR in the parkinsonian animal model. This study suggests that MPE may contain unidentified antiparkinsonian compounds in addition to L -DOPA, or it may have adjuvants that enhance the efficacy of L -DOPA. © 1997 John Wiley & Sons, Ltd.     

论中医现代化应当具备的有效突破

中医现代化的组成部分主要包括理论体系,诊疗技术和中药现代化[1].多数学者认为中西医结合理论体系的建立(或称中医学理论的现代重构)是一个再造中医学术的长远目标[2],从自然哲学到自然科学,从经验医学时代跨越实验医学到整体医学时代,无不充满漫道雄关.尽管如此,国内尚有不少学者从不同角度不同方面作了许多有益的探索.大量的事实已经证明,随着中医学的不断发展,在许多方面将面临突破,朝着现代化的目标迈进.笔者认为理论体系的现代化这一中医现代化的核心问题,已经到了非解决不可的时候了,寻找和把握在中医现代化方面的有效突破,对于本学科的发展将起积极的推进作用.笔者就可能发生的几方面的突破展开讨论.     

雷公藤甲素对BRL大鼠肝细胞增殖和凋亡的影响

目的研究雷公藤甲素(TP)对BRL大鼠肝细胞增殖和凋亡的影响。方法体外培养BRL大鼠肝细胞,经不同浓度TP处理细胞后,采用CCK-8法检测BRL大鼠肝细胞增殖能力;生化仪测定细胞上清液中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)活性;AV/PI双染后流式细胞仪检测细胞的凋亡率;酶联免疫吸附测定法(ELISA)测定细胞上清液中肿瘤坏死因子(TNF-α)、白介素10(IL-10)、干扰素-γ(IFN-γ)等细胞因子含量。结果与正常对照组比较,雷公藤甲素能明显抑制BRL大鼠肝细胞增殖,诱导细胞凋亡;雷公藤甲素1、10、100μmol/L能明显升高BRL大鼠肝细胞上清液中AST活性,100μmol/L能明显升高ALT活性并降低IL-10含量(P0.05,P0.01)。与雷公藤甲素100μmol/L组比较,雷公藤甲素1、10μmol/L组细胞上清液中ALT含量明显降低,雷公藤甲素10μmol/L组细胞上清液中IL-10含量明显升高,雷公藤甲素1μmol/L组细胞凋亡率明显降低且细胞上清液中TNF-α含量明显升高(P0.05,P0.01)。与雷公藤甲素10μmol/L组比较,雷公藤甲素1μmol/L组细胞凋亡率明显降低,细胞上清液中TNF-α含量明显升高(P0.01)。结论雷公藤甲素对BRL大鼠肝细胞增殖具有明显的抑制作用,且呈浓度依赖性,该抑制作用可能与诱导BRL大鼠肝细胞凋亡有关。     

Vandellia cordifolia regulated cell proliferation and cytokines production in human mononuclear cells

Vandellia cordifolia (V. cordifolia) used for treatment inflammation in traditional Chinese medicine was selected for immunopharmacological activity test. The effects of V. cordifolia extracted fractions on human mononuclear cells (HMNC) proliferation were determined by tritiated thymidine uptake. The results indicated that VC-ME fraction suppressed HMNC proliferation activated with phytohemagglutinin (PHA) and stimulated cell cycle progression was arrested at the G0/G1 stage. The inhibitory mechanisms may involve the blocking of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) production, since VC-ME suppressed IL-2 and IFN-gamma production of HMNC in a dose-dependent manner. Therefore, it is suggested that immunomodulatory agents are contained in V. cordifolia.     

Corydalis yanhusuo W.T. Wang extract inhibits MCF-7 cell proliferation by inducing cell cycle G2/M arrest

Corydalis yanhusuo W.T. Wang (YHS) is a traditional Chinese herb widely prescribed for promoting blood circulation, reinforcing vital energy and alleviating pain. Our previous studies showed that an ethanol extract of YHS inhibits metastasis of breast cancer cells in vitro. In the present study, the anti-proliferative effect of the extract was determined by MTT assay and the LDH release was measured with a commercial kit. Intracellular reactive oxygen species (ROS) production and mitochondrial membrane potential (ΔΨm) were monitored by CM- H(2)DCF-DA and JC-1 staining, respectively. Cell cycle was analyzed with propidium iodide (PI) staining by flow cytometry and protein expressions were measured by Western blotting. The YHS extract significantly inhibited MCF-7 cell proliferation in a dose-dependent manner. Significant increase of ROS formation and decrease of ΔΨm were observed. Furthermore, it induced MCF-7 cell cycle arrest at the G2/M phases. In addition, the p-cdc-2/cdc-2 protein expression ratio was increased while Rb and p21 protein expressions were decreased. The YHS extract inhibited MCF-7 proliferation by inducing G2/M cell cycle arrest, which might be mediated by inducing ROS formation, decreasing ΔΨm and regulating cell cycle related protein expressions.     

总丹参酮对HSC-T6细胞增殖和产生胶原的影响

目的 研究总丹参酮对体外肝星状细胞(HSC-T6)增殖和胶原合成的影响.方法 采用血清刺激HSc-T6细胞,用[3H]-TdR和[3H]-Proline掺入法分别检测其增殖和胶原合成的活性.结果 总丹参酮在1.0～16.0 mg·L<'-1>浓度范围内能够浓度依赖性地抑制血清促进的HSC-T6细胞增殖和产生胶原.结论 总丹参酮对HSC-T6细胞的增殖和产生胶原有明显的抑制作用.该作用可能是总丹参酮抗肝纤维化的机理之一.     

芦荟大黄素对Jurkat T细胞增殖和凋亡的作用及机制

目的 研究芦荟大黄素对Jurkat T细胞的增殖抑制和凋亡诱导作用,并探讨可能的机制.方法 应用细胞计数测定增殖,Hoechst/PI双染法及DNA片断化分析细胞凋亡特征,流式细胞术检测细胞周期分布.进一步应用二氢乙锭荧光染色测量活性氧产物,Western blotting检测胞浆细胞色素C(Cyt-c)的量,比色法检测caspase-3和caspase-9活性.结果 芦荟大黄素剂量和时间依赖性地抑制Jurkat T细胞增殖.芦荟大黄素诱发细胞核皱缩,核DNA片断化,细胞周期出现亚G1期凋亡峰,且停滞于G2/M期.芦荟大黄素介导细胞内活性氧水平显著提高,线粒体释放Cyt-c量显著增加,caspase-3和caspase-9活性显著增强.结论 芦荟大黄素剂量依赖性地抑制Jurkat T细胞增殖并诱导其凋亡.诱导凋亡的机制中,包括增加活性氧产物和线粒体损伤途径.     

Yellow tea is more potent than other types of tea in suppressing liver toxicity induced by carbon tetrachloride in rats

The present study compared the effects of six Chinese teas categorized by their production process: green, white, yellow, oolong, black and pu-erh teas, on carbon tetrachloride (CCl4)-induced liver injury. Wistar rats were given ad libitum the Chinese teas prepared according to the home-style methods for 1 week, and then intraperitoneally injected with CCl4 (1 mg/kg body weight) or olive oil as a vehicle. The yellow tea significantly ameliorated the increase in the activity of the alanine- and aspartate-aminotransferases in plasma. Thus, the drinking of yellow tea may contribute to protection against liver injury.     

山茱萸、生地有效部位及配伍组合抑制糖基化终末产物致肾小球系膜细胞增殖的研究

目的:通过体外培养大鼠肾小球系膜细胞(GMC),建立糖基化终末产物(AGEs)致肾小球系膜细胞增殖的细胞模型,观察山茱萸有效部位、生地有效部位及其配伍组合对糖基化终末产物致肾小球系膜细胞增殖的抑制作用,以确定最佳配伍组合物。方法:体外培养肾系膜细胞,当细胞贴壁生长至80%~90%时,(1),用浓度为50、100、200、400mg/L的山茱萸各部位(环烯醚萜苷、三萜酸、多糖)、生地各部位(环烯醚萜苷、多糖、寡糖)进行干预,然后加入200mg/L的糖基化终末产物刺激,24 h后检测细胞增殖情况,筛选出活性较强的部位。(2),将活性较强的部位山茱萸环烯醚萜苷和三萜酸(A)、生地环烯醚萜苷(B)按1:0、0:1、1:1、1:2、2:1进行配伍组合(分别为A1、B1、A1B1、A1B2、A2B1),以浓度为25、50、100mg/L生药及含药血清进行干预,加入200mg/L的AGEs刺激,24 h后检测细胞增殖情况以及细胞上清液中纤维连接蛋白(FN)、层粘连蛋白(LN)、Ⅳ型胶原(Coll-Ⅳ)水平。结果:(1)山茱萸环烯醚萜苷部位、山茱萸三萜酸部位与生地环烯醚萜苷部位活性较强,具有显著抑制糖基化终末产物致肾小球系膜细胞增殖的作用。(2)配伍组合物(A1、B1、A1B1、A1B2、A2B1)生药及含药血清均能够抑制肾小球系膜细胞增殖及其分泌纤维连接蛋白、层粘连蛋白、Ⅳ型胶原的水平,并呈一定量效关系。结论:山茱萸环烯醚萜苷部位、山茱萸三萜酸部位、生地环烯醚萜苷部位及其配伍组合能够明显抑制糖基化终末产物致肾系膜细胞增殖及其分泌的纤维连接蛋白、层粘连蛋白、Ⅳ型胶原水平,减少细胞外基质沉积,其中配伍组合A2B1抑制效应最强,为最佳配伍组合。     

中药有效成分防治骨关节炎的作用机制研究进展

骨关节炎是临床上最常见的关节退行性疾病,影响整个关节组织。中药因低不良反应、低成本、多靶点起效等优势在骨关节炎的防治中占据重要位置。研究发现中药有效成分防治骨关节炎的作用机制主要包括减少炎症因子的产生、减轻氧化应激、抑制基质降解酶、调节软骨细胞代谢及调控相关信号通路等方面。主要综述了中药有效成分防治骨关节炎作用机制的研究进展,旨在为进一步研究中药有效成分在骨关节炎防治中的应用价值提供参考和依据。     

Constituents isolated from Cordyceps militaris suppress enhanced inflammatory mediator's production and human cancer cell proliferation

Aim of the study

The purpose of this study is to isolate the pure compounds from the extracts of Cordyceps militaris obtained through solid-state cultivation process, and evaluate their anti-inflammatory and anticancer properties.

Materials and methods

Silica gel column chromatographic purification of Cordyceps militaris extracts resulted in the isolation of 10 pure compounds (1-10). The compounds 1-10 were examined for their growth inhibitory properties against nitric oxide (NO), tumor necrosis factor (TNF)-α and interleukin (IL)-12 enhanced production from LPS/IFN-γ-stimulated macrophages. Additionally, the anti-proliferation effects of 1-10 on human cancer cell lines, colon (colon 205), prostate (PC-3), and hepatoma (HepG2) cells were also analyzed.

Results

Compound 8 displayed potent growth inhibition on NO, TNF-α and IL-12 production with an IC₅₀ value of 7.5, 6.3, and 7.6 μg/ml, respectively. A similar inhibitory trend on these inflammatory mediators was observed for 3, 7, 9 and 10 with an IC₅₀ values ranging from 10.8 to 17.2 μg/ml. On the other hand, compounds 3 and 8 were potent anti-proliferative agents with an IC₅₀ value of 35.6 and 32.6 μg/ml toward PC-3 and colon 205 cell lines, respectively. The compounds 1 and 2 showed potent anti-proliferation in PC-3 and colon 205 cells, while only 3 displayed such effect in HepG2 cells.

Conclusion

The present study provides scientific supporting information for the ethnopharmacological use of Cordyceps militaris as an anti-inflammatory and anticancer agent.     

雷公藤属3种植物不同群体和个体中雷公藤甲素的研究

目的测定雷公藤属3种植物(雷公藤、昆明山海棠和黑蔓)不同群体和个体中雷公藤甲素(tripto lide),为评价雷公藤药材质量和寻找雷公藤的优质种质奠定基础。方法建立雷公藤甲素HPLC测定方法,并测定了全国主要分布区25个群体91个个体的木质部和韧皮部中的雷公藤甲素。结果黑蔓的雷公藤甲素质量分数很低;雷公藤和昆明山海棠种间的雷公藤甲素差异不明显,个体质量分数木质部为1.0×10-6~5.83×10-5;韧皮部为2.3×10-6~1.030×1-0 4。结论雷公藤和昆明山海棠不同个体雷公藤甲素质量分数最高值与最低值相差约50倍,不同居群雷公藤甲素质量分数最高值与最低值相差10多倍,不同来源的药材质量差异极大,严重影响用药的安全性;雷公藤甲素质量分数高的居群位于浙江西南部和中部、湖南新宁、贵州雷山和安徽黄山;湖南新宁、贵州雷山和浙江江山居群中有雷公藤甲素质量分数极高的个体,值得进一步研究,以寻找质量分数高的优良单株。     

淫羊藿素和脱水淫羊藿素对人类乳腺癌细胞T47D增殖和细胞周期的影响

目的 探讨淫羊藿素和脱水淫羊藿索对人类乳腺癌细胞株T47D增殖和细胞周期的影响.方法 采用噻唑蓝(MTT)比色法测定淫羊藿素和脱水淫羊藿素对雌激素依赖性乳腺癌细胞T47D的细胞增殖作用.并以雌激素受体拮抗剂ICI 182,780和雌激素受体ERB激动剂DPN为工具药来评价淫羊藿素和脱水淫羊藿素发挥雌激素样作用与雌激素受体的关系,流式细胞术对T47D细胞的增殖情况进行分析.结果 淫羊藿素和脱水淫羊藿素在10-8～10-6范围内能促进T47D细胞的增殖,并将T47D细胞周期由G1期向S期推进,促进DNA合成,提高细胞分裂增殖指数,且淫羊藿素和脱水淫羊藿素促进T47D细胞增殖作用被雌激素受体拮抗剂所拮抗.结论 淫羊藿素和脱水淫羊藿素具有雌激素活性,此作用可能是通过雌激素受体(ER)介导的.     

吉西他滨对人膀胱癌细胞株T24增殖、凋亡及自噬相关蛋白表达的影响

目的观察吉西他滨对人膀胱癌细胞株T24增殖、凋亡及自噬相关基因表达的影响。方法将T24细胞分为阳性对照组、低剂量实验组、中剂量实验组、高剂量实验组和空白对照组;阳性对照组用10~(-8) mol/L紫杉醇处理,低、中、高剂量实验组以1×10~(-7),1×10~(-8),1×10~(-9) mol/L吉西他滨处理,空白对照组不做任何处理。药物干预24,48 h后采用溴化四唑蓝(MTT)法检测细胞增殖情况,流式细胞术检测细胞凋亡情况,蛋白免疫印迹法(WB)检测细胞中自噬相关蛋白表达情况,透射电镜扫描细胞中自噬小体的形成情况。结果与阳性对照组比较,低、中剂量实验组培养24,48 h增殖抑制率(PIR)均明显降低(P均0.05);实验组T24细胞PIR均随着药物浓度的增加和培养时间的延长而增高(P均0.05)。阳性对照组和低、中、高剂量实验组细胞凋亡率均显著高于空白对照组(P均0.05)。随着培养时间的延长,中剂量实验组自噬相关蛋白蛋白表达量呈逐渐升高趋势(P0.05),P62蛋白表达量呈逐渐减低趋势(P0.05)。培养4 h后,中剂量实验组细胞中出现了较多的自噬小体。结论吉西他滨可抑制T24细胞增殖,并促其凋亡,具有一定时间-剂量依赖性;同时其还可诱导细胞自噬的发生。     

雷公藤药材中雷公藤甲素及总二萜内酯的含量测定

目的 建立雷公藤药材中雷公藤甲素及总二萜内酯含量测定的方法.方法 采用高效液相色谱法,色谱柱：Platisil ODS柱（250mm×4.6mm,5μm）,以甲醇-水（45∶55）为流动相进行洗脱,于218nnm波长处检测,测定雷公藤甲素的含量;采用Kedde显色法,在550nm波长处测定吸光度,测定总二萜内酯的含量.结果 以高效液相色谱法测定雷公藤甲素,在0.58～46.40μg·mL-1（r=0.9999）范围内线性关系良好,平均回收率为100.52％ （n =6）;比色法测定雷公藤总二萜内酯（以雷公藤甲素计）在0.49-39.36μg·mL-1（r =0.9999）范围内线性关系良好,平均回收率为93.28％（n=6）;雷公藤药材中雷公藤甲素平均含量为4.20μg·g-1,总二萜内酯为86.64μg·g-1.结论 本实验所建立的方法稳定,精密度良好,适合于雷公藤甲素及总二萜内酯的含量测定,可用于雷公藤药材的质量控制.     

Auricular acupuncture increases cell proliferation in the dentate gyrus of Sprague-Dawley rats.

In a previous study, we have reported that acupuncture increases cell proliferation in the dentate gyrus after transient global ischemia in gerbils. In the present study, the effect of auricular acupuncture (AA) on dentate cell proliferation was examined in appropriately fed and food-deprived adult Sprague-Dawley (S-D) rats. 5-Bromo-2'-deoxyuridine-5'-monophosphate (BrdU) immunohistochemistry revealed a significant increase in cell birth in the dentate gyrus of both groups after AA treatment. In addition, the effect of AA on cell proliferation in the dentate gyrus was more profound in the food-supplied group. These findings demonstrate that AA enhances cell proliferation in the dentate gyrus of adult rats.     

胰岛素抵抗3T3-L1脂肪细胞模型及中药有效成分的干预作用

高糖高胰岛素、游离脂肪酸、炎症因子、地塞米松、金属铬、雌激素、葡糖胺等均可诱导3T3-L1脂肪细胞建立胰岛素抵抗模型,且具有稳定、可靠、易于重复等优点。目前已证实小檗碱+梓醇、小檗碱、熊果酸、西洋参茎叶总皂苷及其他活性成分、蒲黄总黄酮、灵芝多糖、附子多糖、地黄寡糖、积雪草酸、白藜芦醇、葛根素等中药有效成分具有改善胰岛素抵抗及调节糖代谢作用。