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Chinese Journal of Natural Medicines 2016年第3期14卷 203-209页

作者： lin sen-sen LI Fang-Fang SUN Li FAN Wei GU Ming ZHANG Lu-Yong QIN Song YUAN Sheng-Tao New Drug Screen Center China Pharmaceutical University Nanjing Sanhome Pharmaceutical Co. Ltd.

Marsdenia tenacissima, a traditional Chinese medicine, is long been used to treat various diseases including asthma, cancer, trachitis, tonsillitis, pharyngitis, cystitis, and pneumonia. Although Marsdenia tenacissima has been demonstrated to have strong anti-tumor effects against primary tumors, its effect on cancer metastasis remains to be defined, and the molecular mechanism underlying the anti-metastatic effect is unknown. In the present study, we investigated the effects of XAP(an extract of Marsdenia tenacissima) on A549 lung cancer cell migration and explored the role of CCR5-CCL5 axis in the anti-metastatic effects of XAP. Our resutls showed that XAP inhibited A549 lung cancer cell migration and invasion in a dose-dependent manner. The protein levels of CCR5, but not CCR9 and CXCR4, were decreased by XAP. The secretion of CCL5, the ligand of CCR5, was reduced by XAP. XAP down-regulated Rho C expression and FAK phosphorylation. In conclusion, XAP inhibited A549 cell migration and invasion through down-regulation of CCR5-CCL5 axis, Rho C, and FAK.

关键词： XAP A549 cell migration CCR5-CCL5 axis Rho C FAK

Potent anti-angiogenicactivity of B19—a mono-carbonyl analogue of curcumin

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Chinese Journal of Natural Medicines 2014年第1期12卷 8-14页

作者： SUN Li LIU Jin lin sen-sen SHI Wen-Ting ZHU Jing LIANG Guang YUAN Sheng-Tao Jiangsu Center for Drug Screening China Pharmaceutical University Nanjing 210009 China Jiangsu Center for Pharmacodynamics Research and Evaluation China Pharmaceutical University Nanjing 210009 China Wenzhou Medical College Pharmacy School Wenzhou 325035 China

AIM： The compound B19 （C21H22O5） is a newly synthesized, mono-carbonyl analog of curcumin that has exhibited potentialantitumor effects. This present study was performed to identify the anti-angiogenic activity of this compound. METHODS AND RESULTS： B19 inhibited migration and tube formation of human umbilical vein endothelial cells, and arrested microvessel outgrowth from rat aortic *** addition, B19 suppressed the neovascularization of chicken chorioallantoic membrane. Mechanistic studies revealed that B19 suppressed the downstream protein kinase activation of vascular endothelial growth factor （VEGF） by decreasing phosphorylated forms of serine/threonine kinase Akt, extracellular signal-regulated kinase, and p38 mitogen-activated protein kinase, with or without stimulating vascular endothelial growth factor （VEGF）. CONCLUSIONS： B 19 exerted anti-angiogenic activity in vitro and ex vivo, which suggests that it merits further investigation as a promising anticancer angiogenesis compound.

关键词： B 19 Curcumin derivative Angiogenesis Human umbilical vein endothelial cell (HUVEC) Vascular endothelial growth factor (VEGF)

Marsdenia tenacissima extract induces G_0/G_1 cell cycle arrest in human esophageal carcinoma cells by inhibiting mitogen-activated protein kinase(MAPK) signaling pathway

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Chinese Journal of Natural Medicines 2015年第6期13卷 428-437页

作者： FAN Wei SUN Li ZHOU Jing-Qian ZHANG Cang QIN Song TANG Ying LIU Yang lin sen-sen YUAN Sheng-Tao Jiangsu Center for Drug Screening China Pharmaceutical University Department of Complex Prescription of Traditional Chinese Medicine School of Chinese Material Medicine China Pharmaceutical University Nanjing Sanhome Pharmaceutical Co. Ltd. Jiangsu Center for Pharmacodynamics Research and Evaluation China Pharmaceutical University

Marsdenia tenacissima extract(MTE, trade name: Xiao-Ai-Ping injection) is an extract of a single Chinese plant medicine. It has been used for the treatment of cancer in China for decades, especially for esophageal cancer and other cancers in the digestive tract. In the present study, the potential mechanism for MTE's activity in esophageal cancer was explored. The effects of MTE on the proliferation of human esophageal cancer cells(KYSE150 and Eca-109) were investigated by the MTT assay, the Brd U(bromodeoxyuridine) incorporation immunofluorescence assay, and flow cytometric analysis. MTE inhibited cell proliferation through inducing G0/G1 cell cycle arrest in KYSE150 and Eca-109. Western blot analysis was employed to determine protein levels in the MTE treated cells. Compared with the control cells, the expression levels of the cell cycle regulatory proteins cyclin D1/D2/D3, cyclin E1, CDK2/4/6(CDK: cyclin dependent kinase), and p-Rb were decreased significantly in the cells treated with MTE at 40 mg·m L-1. In addition, MTE had an inhibitory effect on the MAPK(mitogen-activated protein kinase) signal transduction pathway, including ERK(extracellular signal-regulated kinase), JNK(c-Jun N-terminal kinase), and p38 MAPK. Moreover, MTE showed little additional effects on the regulation of cyclin D1/D3, CDK4/6, and p-Rb when the ERK pathway was already inhibited by the specific ERK inhibitor U0126. In conclusion, these data suggest that MTE inhibits human esophageal cancer cell proliferation through regulation of cell cycle regulatory proteins and the MAPK signaling pathways, which is probably mediated by the inhibition of ERK activation.

关键词： Marsdenia tenacissima extract Cell cycle arrest Mitogen-activated protein kinase signaling pathway Human esophageal cancer

DT-13, a saponin of dwarf lilyturf tuber, exhibits anti-cancer activity by down-regulating C-C chemokine receptor type 5 and vascular endothelial growth factor in MDA-MB-435 cells

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Chinese Journal of Natural Medicines 2014年第1期12卷 24-29页

作者： ZHAO Ren-Ping lin sen-sen YUAN Sheng-Tao YU Bo-Yang BAI Xian-Shu SUN Li ZHANG Lu-Yong Jiangsu Center for Drug Screening China Pharmaceutical University Jiangsu Center for Pharmacodynamics Research and Evaluation China Pharmaceutical University Department of Complex Prescription of Traditional Chinese Medicine School of Chinese Material Medicine China Pharmaceutical University Department of Molecular Physiology University of Saarland Key Laboratory of Drug Quality Control and Pharmacovigilance China Pharmaceutical University Ministry of Education

AIM： To investigate the anticancer activity of DT-13 under normoxia and determine the underlying mechanisms of action. METHODS： MDA-MB-435 cell proliferation, migration, and adhesion were performed to assess the anticancer activity of DT-13, a saponin from Ophiopogonjaponicus, in vitro. In addition, the effects of DT-13 on tumor growth and metastasis in vivo were evaluated by orthotopic implantation of MDA-MB-435 cells into nude mice; mRNA levels of vascular endothelial growth factor （VEGF）, C-C chemokine receptor type 5 （CCR5） and hypoxia-inducible factor 1a （HIF-1a） were evaluated by real-time quantitative PCR; and CCR5 protein levels were detected by Western blot assay. RESULTS： At 0.01 to 1 umol·L -1, DT-13 inhibited MDA-MB-435 cell proliferation, migration, and adhesion significantly in vitro. DT-13 reduced VEGF and CCR5 mRNAs, and decreased CCR5 protein expression by down-regulating HIF-1 a. In addition, DT-13 inhibited MDA-MB-435 cell lung metastasis, and restricted tumor growth slightly in vivo. CONCLUSION： DT-13 inhibited MDA-MB-435 cell proliferation, adhesion, and migration in vitro, and lung metastasis in vivo by reducing VEGF, CCR5, and HIF-la expression.

关键词： DT- 13 Saponin Ophiopogonjaponicus Anticancer activity CCR5 VEGF HIF- 1 a

The saponin DT-13 inhibits gastric cancer cell migration through down-regulation of CCR5-CCL5 axis

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Chinese Journal of Natural Medicines 2014年第11期12卷 833-840页

作者： lin sen-sen FAN Wei SUN Li LI Fang-Fang ZHAO Ren-Ping ZHANG Lu-Yong YU Bo-Yang YUAN Sheng-Tao New Drug Screen Center China Pharmaceutical University Department of Complex Prescription of TCM China Pharmaceutical University

AIM: To investigate the effect of DT-13 on gastric cancer cell migration, and to explore the possible mechanisms underlying the anti-metastasis activity of DT-13. METHODS: Growth inhibition of DT-13 was analyzed by the MTT assay. Cell migration was measured by the scratch-wound assay and transwell double chamber assay. To investigate the possible mechanisms underlying the anti-metastasis activity of DT-13, chemokine receptors that are involved in cancer metastasis(CCR2, CCR5, CCR7, CXCR4, and CXCR6) were detected by conventional PCR. The effect of DT-13 on CCR5 and CXCR4 expression was further evaluated by quantitative PCR and Western blot, respectively. The secretion of CCL5(ligand of CCR5) and SDF-1(ligand of CXCR4) were detected by enzyme-linked immunosorbent assay(ELISA). RESULTS: DT-13 inhibited BGC-823 and HGC-27 cell growth in a dose dependent manner, and the estimated IC50 value for 24 h treatment was 23.5 ± 5.1 μmol·L-1 for BGC-823 cells and 35.6 ± 7.6 μmol·L-1 for HGC-27 cells. DT-13 also significantly decreased gastric cancer cell migration. DT-13 significantly decreased the gene expression of CCR5 in both BGC-823 and HGC-27 gastric cancer cells, and moderately reduced the expression of CXCR4. Similar to the results of gene expression, significant down-regulation of CCR5 protein was observed, but CXCR4 protein levels were much less affected. CCL5 secretion, but not SDF-1 production, was inhibited by DT-13. CONCLUSION: DT-13 inhibited gastric cancer cell migration by down-regulation of the CCR5-CCL5 axis.

关键词： Liriope muscari Saponin DT-13 BGC-823 HGC-27 Gastric cancer cell migration CCR5 CCL5 CXCR4

复合保鲜剂对腐生葡萄球菌的抑菌活性及其作用机理

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食品与机械 2018年第4期34卷 158-162页

作者：朱亚珠林森森浙江国际海运职业技术学院浙江舟山316021

以山梨酸钾、双乙酸钠和异抗坏血酸钠作为复合保鲜剂的基本成分,研究其对虾皮中腐生葡萄球菌的抑菌作用。通过酶标法来确定复合保鲜剂对腐生葡萄球菌的最小抑菌浓度(MIC),测定抑菌活力、细菌生长曲线、细胞膜完整性、碱性磷酸酶(AKP)含... 详细信息

以山梨酸钾、双乙酸钠和异抗坏血酸钠作为复合保鲜剂的基本成分,研究其对虾皮中腐生葡萄球菌的抑菌作用。通过酶标法来确定复合保鲜剂对腐生葡萄球菌的最小抑菌浓度(MIC),测定抑菌活力、细菌生长曲线、细胞膜完整性、碱性磷酸酶(AKP)含量等指标,并结合细菌超微结构观察,综合评价复合保鲜剂对腐生葡萄球菌的作用效果。结果表明:复合保鲜剂能明显降低腐生葡萄球菌的生长速率,其最低抑菌浓度(MIC)为109μg/mL,最低杀菌浓度(MBC)为219μg/mL,抑菌活力均在使用初期的作用最强(约3h);随着作用时间的延长,菌液中的碱性磷酸酶量显著增多,在260nm处的吸收值显著增加,表明菌体内部核酸与蛋白质外泄,菌体细胞壁与细胞膜受到破坏。此外,电镜扫描结果显示,试验组的菌体发生干瘪破裂现象,细胞质从细胞内大量渗出。故一定浓度的复合保鲜剂对虾皮中的腐生葡萄球菌有抑制(杀灭)作用,对细胞体有一定破坏作用。

关键词：复合保鲜剂腐生葡萄球菌抑菌机理

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内皮细胞三维培养芽生模型的构建

中国药科大学学报 2012年第1期43卷 60-63页

作者：王萍孙立林森森袁胜涛中国药科大学新药筛选中心南京210009

构建内皮细胞三维芽生模型,为调控肿瘤血管生成药物的药效研究提供一种更为准确的检测手段。通过倒挂悬滴式的培养方法使脐静脉内皮细胞形成致密的细胞团,再将细胞团分别放入Matrigel,鼠尾胶及纤维胶3种基质中进行三维培养。结果显示内... 详细信息

构建内皮细胞三维芽生模型,为调控肿瘤血管生成药物的药效研究提供一种更为准确的检测手段。通过倒挂悬滴式的培养方法使脐静脉内皮细胞形成致密的细胞团,再将细胞团分别放入Matrigel,鼠尾胶及纤维胶3种基质中进行三维培养。结果显示内皮细胞团在纤维胶中形成了明显的相互连接的毛状管腔样结构,即芽生。经验证,内皮细胞团在纤维胶中能形成明显的管腔样结构,且该方法能够成为一种体外观察研究管腔形成的有效方法。

关键词：血管生成内皮细胞芽生细胞培养三维体系

UPLC-Q-TOF-MS分析鸡血藤乙酸乙酯提取物中抑制表皮生长因子受体活性成分

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北京中医药大学学报 2014年第1期37卷 58-62页

作者：王冬梅林森森郑司浩黄林芳中国医学科学院药用植物研究所北京100193 中国药科大学新药筛选中心

目的应用均相时间分辨荧光（HTRF）技术，筛选发现具有抑制表皮生长因子受体（EGFR）活性的鸡血藤提取物，同时采用超高效液相色谱．飞行时间质谱联用技术（UPLC—Q—***）对其活性部位进行分析和鉴别。方法药材经石油醚渗滤、乙醇提取... 详细信息

目的应用均相时间分辨荧光（HTRF）技术，筛选发现具有抑制表皮生长因子受体（EGFR）活性的鸡血藤提取物，同时采用超高效液相色谱．飞行时间质谱联用技术（UPLC—Q—***）对其活性部位进行分析和鉴别。方法药材经石油醚渗滤、乙醇提取、乙酸乙酯萃取和水煎煮后，得到4个提取部位。采用HTRF法检测鸡血藤乙酸乙酯提取物对EGFR的抑制作用，计算提取物对EGFR的抑制率。采用ACQUITYUPLCBEH C-18色谱柱，以0．1％甲酸水溶液（A）．乙腈（B）为流动相梯度洗脱，200～400nm扫描，使用ESI离子源，在正离子模式下采集数据。结果首次发现鸡血藤的乙酸乙酯部位显示较强的EGFR抑制活性，IC-18为2．277mg／L，并从其活性部位中分析鉴定出15个化学物，主要成分为黄酮类化合物，其中10个分别是儿茶素．儿茶素，阿福豆素一儿茶素，杨梅素-3＇-O—β—D吡喃木糖苷，芒柄花苷，芒柄花黄素，Afrormosin，表没食子儿茶素，催吐萝芙木醇-9-O-木糖-（1→6）葡萄糖苷，木犀草苷，没食子儿茶素．儿茶素；5个未知成分。儿茶素、表没食子儿茶素、芒柄花黄素为主要活性成分。结论鸡血藤乙酸乙酯提取物所含的黄酮类成分为抑制EGFR活性的主要成分，本研究以期为其在抗癌方面的应用提供理论依据，同时为进一步跟踪分离活性成分奠定基础。

关键词：鸡血藤表皮生长因子受体抑制剂均相时间分辨荧光有效成分超高效液相色谱-飞行时间质谱联用技术

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水利工程施工过程中防渗技术分析

工程建设与设计 2024年第21期 107-109页

作者：林森森广东省水利水电第三工程局有限公司广东东莞523000

总结了防渗技术的作用及防渗技术选择原则,分析了高压喷射灌浆、劈裂灌浆、防渗墙(混凝土或搅拌桩)、复合土工膜等防渗技术在水利工程的应用要点。最后,以某水库大坝为研究对象,分析了其防渗方案的施工要点,并以透水率为评价指标,评价... 详细信息

总结了防渗技术的作用及防渗技术选择原则,分析了高压喷射灌浆、劈裂灌浆、防渗墙(混凝土或搅拌桩)、复合土工膜等防渗技术在水利工程的应用要点。最后,以某水库大坝为研究对象,分析了其防渗方案的施工要点,并以透水率为评价指标,评价了大坝的防渗效果。

关键词：水利工程防渗原则防渗技术施工案例防渗效果

全长及细胞内磷酸化位点缺失的人组织因子真核表达重组质粒的构建及其生物学功能

同方期刊数据库博看期刊评论

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中国药科大学学报 2009年第5期40卷 465-470页

作者：施伟孙立林森森袁胜涛张陆勇中国药科大学新药筛选中心江苏省药效研究与评价服务中心南京210009

目的:研究组织因子(tissue factor,TF)细胞内区域的功能,分析TF细胞内区域对人乳腺癌细胞MCF-7迁移能力的影响。方法:Trizol一步法抽提人乳腺癌细胞MDA-MB-231总RNA,RT-PCR扩增TF全长(TF full-length)基因和TF细胞内磷酸化位点缺失(TF t... 详细信息

目的:研究组织因子(tissue factor,TF)细胞内区域的功能,分析TF细胞内区域对人乳腺癌细胞MCF-7迁移能力的影响。方法:Trizol一步法抽提人乳腺癌细胞MDA-MB-231总RNA,RT-PCR扩增TF全长(TF full-length)基因和TF细胞内磷酸化位点缺失(TF truncated)基因,分别克隆入pGEM-T载体,再构建人TF full-length基因和TF truncated基因的真核表达质粒pcDNA3.1(+)-TF full-length和pcDNA3.1(+)-TF truncated。将构建好的pcDNA3.1(+)-TF full-length和pcDNA3.1(+)-TF truncated瞬时转染MCF-7细胞,RT-PCR和Western blotting鉴定重组质粒的表达,并用发色底物法检测瞬时转染后TF的促凝血功能。利用Transwell小室检测转入人TF full-length基因和TF truncated基因后MCF-7细胞的迁移能力。结果:扩增出人TF full-length基因和TF truncated基因,成功克隆入真核表达载体pcDNA3.1(+),并且在MCF-7细胞中表达后具有促凝血活性。瞬时转染pcDNA3.1(+)-TF truncated不能增加MCF-7细胞的迁移能力,而转染pcDNA3.1(+)-TF full-length后可以显著增强MCF-7细胞的迁移能力。结论:成功构建了带有人TF full-length,TF truncated基因的真核表达质粒pcDNA3.1(+)-TF full-length和pcDNA3.1(+)-TF truncated,为研究TF以及其细胞内磷酸化位点区域在肿瘤学中的作用奠定了基础。

关键词：组织因子磷酸化位点缺失真核表达载体pcDNA3.1(+) 细胞迁移