目的探讨虎眼万年青总皂苷(Ornithogalum caudatum AIT saponins)诱导人乳腺癌MDA-MB-231细胞的凋亡的机制,为开发应用于治疗乳腺癌的药物提供实验依据。方法用MTT比色法检测虎眼万年青总皂苷对人乳腺癌MDA-MB-231细胞增殖的抑制作用;...目的探讨虎眼万年青总皂苷(Ornithogalum caudatum AIT saponins)诱导人乳腺癌MDA-MB-231细胞的凋亡的机制,为开发应用于治疗乳腺癌的药物提供实验依据。方法用MTT比色法检测虎眼万年青总皂苷对人乳腺癌MDA-MB-231细胞增殖的抑制作用;流式细胞术分析细胞凋亡;Western Blotting法检测细胞凋亡相关蛋白Bcl-2、Bax与Bad表达的变化。结果虎眼万年青总皂苷对人乳腺癌MDA-MB-231细胞生长有明显抑制作用,并呈剂量、时间依赖性;能诱导人乳腺癌MDA-MB-231细胞发生凋亡,且凋亡率呈浓度依赖性;能下调Bcl-2蛋白的表达,升高Bax/Bcl-2的比值,其高剂量组具有显著统计学差异(P<0.01),能上调Bad蛋白的表达,其低、中、高剂量组具有显著统计学差异(P<0.01)。结论虎眼万年青总皂苷可抑制乳腺癌MDA-MB-231细胞增殖,并可诱导细胞凋亡,其机制可能与升高Bax/Bcl-2的比值,上调Bad蛋白的表达相关。展开更多
OBJECTIVE To investigate toll-like receptor 4(TLR4)-related the regulation of Ornithogalum caudatum extract(OCE) on inflammatory responses in lipopolysaccharide(LPS) activated macro.phages.METHODS Primary peritoneal m...OBJECTIVE To investigate toll-like receptor 4(TLR4)-related the regulation of Ornithogalum caudatum extract(OCE) on inflammatory responses in lipopolysaccharide(LPS) activated macro.phages.METHODS Primary peritoneal macrophage,Raw 264.7,and THP-1 were incubated in 96-well plate for 24 h and treated with OCE of the concentration of 0-400 μg/ml for 4 h.The viability of cells was measured by MTT assay.Specific concentrations of OCE were added into the medium of primary peri.toneal macrophage,Raw 264.7,and THP-1,respectively,then following with lipopolysaccharides(LPS).Cells were harvested and the total cellular protein and nuclear protein were extracted,and the protein content was determined using BCA protein assay Kit.The expressions of TLR4,inducible nitric oxide synthase(iNOS),cyclooxygenase 2(COX-2),α-inhibitor of NF-κB(IκB-α) and nuclear factor-κB(NF-κB) were assayed by Western blot.The expressions of interleukin-1α(IL-1α),interleukin-1β(IL-1β),interleukin-18(IL-18),and tumor necrosis factor-α(TNF-α) were measured by RT-PCR.RESULTS The results of MTT showed that OCE has no cytotoxicity in Raw 264.7 cells between 1.56 μg/ml and 400 μg/ml.Compared with normal group,the expressions of TLR4,iNOS,COX-2,NF-κB and IL-1α,IL-1β,IL-18,TNF-α,the level of nitric oxide(NO) were significantly increased by LPS stimulation,while OCE pretreat.ment reduced these increase induced by LPS.However,OCE pretreatment reversed the reduction of IκB-α after LPS stimulation.CONCLUSION OCE might suppress TLR4 expression and block the inflamma.tion process of NF-κB and iNOS,further decrease the expression of COX-2 and inhibit the release of inflammatory factors.展开更多
目的从虎眼万年青中提取、分离水溶性多糖,初步研究其特征和抗肿瘤活性。方法采用热水提取,乙醇沉淀,Sevag法脱蛋白,DEAE-Sepharose Fast Flow离子交换柱色谱和Sephadex G-75凝胶过滤柱色谱分离纯化,得到虎眼万年青均一多糖OCAP-2-2。...目的从虎眼万年青中提取、分离水溶性多糖,初步研究其特征和抗肿瘤活性。方法采用热水提取,乙醇沉淀,Sevag法脱蛋白,DEAE-Sepharose Fast Flow离子交换柱色谱和Sephadex G-75凝胶过滤柱色谱分离纯化,得到虎眼万年青均一多糖OCAP-2-2。毛细管区带电泳法(CZE)分析单糖组成;采用高效凝胶渗透色谱法(HPGPC)测定多糖纯度和相对分子质量;动物移植性实体瘤的瘤重实验法研究对小鼠S180肉瘤的抑瘤作用,采用细胞体外培养技术,MTT法检测对人白血病细胞株K562细胞的增殖抑制作用。结果经过分离纯化得到的均一多糖组分OCAP-2-2主要由葡萄糖、木糖、甘露糖、半乳糖等4种单糖组成,其相对分子质量之比为2.16∶1.26:0.88∶1.00,平均相对分子质量9.84×104,总糖含量为92.3%,总糖醛酸含量为6.21%,蛋白质含量为3.68%;在0.1～100μg.mL-1内与荷瘤对照组相比,OCAP-2-2对小鼠S180肉瘤有显著的抑制活性,其中多糖浓度为100μg.mL-1时抑瘤率达(53.16±4.23)%(P<0.001);OCAP-2-2对K562细胞有明显的增殖抑制作用(P<0.01),在多糖浓度为0.10μg.mL-1时,增殖抑制率最高为(39.83±7.31)%(P<0.01)。结论 OCAP-2-2具有很高的抗肿瘤活性,可以探索作为一种潜在的天然抗肿瘤药物。展开更多
A water soluble crude extract prepared from Ornithogalum caudatum Ait. (OCA) showing a high immunomodulating activitiy was isolated and characterized by virtue of gel filtration and column chromatography. The pre...A water soluble crude extract prepared from Ornithogalum caudatum Ait. (OCA) showing a high immunomodulating activitiy was isolated and characterized by virtue of gel filtration and column chromatography. The presence of the monosaccharides has been established by the chemical analysis. The quantitative analysis of the alditol acetate derivatives of them showed the ratios of the monosaccharides analyzed by means of GC respectively. The concentrations of protein(280 nm) and carbohydrate(496 nm) were detected respectively. The information of the molecular weight from the pure polysaccharide was obtained by several standard Dextrans from the Sephadex chromatography.展开更多
文摘目的探讨虎眼万年青总皂苷(Ornithogalum caudatum AIT saponins)诱导人乳腺癌MDA-MB-231细胞的凋亡的机制,为开发应用于治疗乳腺癌的药物提供实验依据。方法用MTT比色法检测虎眼万年青总皂苷对人乳腺癌MDA-MB-231细胞增殖的抑制作用;流式细胞术分析细胞凋亡;Western Blotting法检测细胞凋亡相关蛋白Bcl-2、Bax与Bad表达的变化。结果虎眼万年青总皂苷对人乳腺癌MDA-MB-231细胞生长有明显抑制作用,并呈剂量、时间依赖性;能诱导人乳腺癌MDA-MB-231细胞发生凋亡,且凋亡率呈浓度依赖性;能下调Bcl-2蛋白的表达,升高Bax/Bcl-2的比值,其高剂量组具有显著统计学差异(P<0.01),能上调Bad蛋白的表达,其低、中、高剂量组具有显著统计学差异(P<0.01)。结论虎眼万年青总皂苷可抑制乳腺癌MDA-MB-231细胞增殖,并可诱导细胞凋亡,其机制可能与升高Bax/Bcl-2的比值,上调Bad蛋白的表达相关。
基金supported by National Natural Science Foundation of China(8146056481760668+3 种基金81560597812606648136065881660689)
文摘OBJECTIVE To investigate toll-like receptor 4(TLR4)-related the regulation of Ornithogalum caudatum extract(OCE) on inflammatory responses in lipopolysaccharide(LPS) activated macro.phages.METHODS Primary peritoneal macrophage,Raw 264.7,and THP-1 were incubated in 96-well plate for 24 h and treated with OCE of the concentration of 0-400 μg/ml for 4 h.The viability of cells was measured by MTT assay.Specific concentrations of OCE were added into the medium of primary peri.toneal macrophage,Raw 264.7,and THP-1,respectively,then following with lipopolysaccharides(LPS).Cells were harvested and the total cellular protein and nuclear protein were extracted,and the protein content was determined using BCA protein assay Kit.The expressions of TLR4,inducible nitric oxide synthase(iNOS),cyclooxygenase 2(COX-2),α-inhibitor of NF-κB(IκB-α) and nuclear factor-κB(NF-κB) were assayed by Western blot.The expressions of interleukin-1α(IL-1α),interleukin-1β(IL-1β),interleukin-18(IL-18),and tumor necrosis factor-α(TNF-α) were measured by RT-PCR.RESULTS The results of MTT showed that OCE has no cytotoxicity in Raw 264.7 cells between 1.56 μg/ml and 400 μg/ml.Compared with normal group,the expressions of TLR4,iNOS,COX-2,NF-κB and IL-1α,IL-1β,IL-18,TNF-α,the level of nitric oxide(NO) were significantly increased by LPS stimulation,while OCE pretreat.ment reduced these increase induced by LPS.However,OCE pretreatment reversed the reduction of IκB-α after LPS stimulation.CONCLUSION OCE might suppress TLR4 expression and block the inflamma.tion process of NF-κB and iNOS,further decrease the expression of COX-2 and inhibit the release of inflammatory factors.
文摘目的从虎眼万年青中提取、分离水溶性多糖,初步研究其特征和抗肿瘤活性。方法采用热水提取,乙醇沉淀,Sevag法脱蛋白,DEAE-Sepharose Fast Flow离子交换柱色谱和Sephadex G-75凝胶过滤柱色谱分离纯化,得到虎眼万年青均一多糖OCAP-2-2。毛细管区带电泳法(CZE)分析单糖组成;采用高效凝胶渗透色谱法(HPGPC)测定多糖纯度和相对分子质量;动物移植性实体瘤的瘤重实验法研究对小鼠S180肉瘤的抑瘤作用,采用细胞体外培养技术,MTT法检测对人白血病细胞株K562细胞的增殖抑制作用。结果经过分离纯化得到的均一多糖组分OCAP-2-2主要由葡萄糖、木糖、甘露糖、半乳糖等4种单糖组成,其相对分子质量之比为2.16∶1.26:0.88∶1.00,平均相对分子质量9.84×104,总糖含量为92.3%,总糖醛酸含量为6.21%,蛋白质含量为3.68%;在0.1～100μg.mL-1内与荷瘤对照组相比,OCAP-2-2对小鼠S180肉瘤有显著的抑制活性,其中多糖浓度为100μg.mL-1时抑瘤率达(53.16±4.23)%(P<0.001);OCAP-2-2对K562细胞有明显的增殖抑制作用(P<0.01),在多糖浓度为0.10μg.mL-1时,增殖抑制率最高为(39.83±7.31)%(P<0.01)。结论 OCAP-2-2具有很高的抗肿瘤活性,可以探索作为一种潜在的天然抗肿瘤药物。
文摘A water soluble crude extract prepared from Ornithogalum caudatum Ait. (OCA) showing a high immunomodulating activitiy was isolated and characterized by virtue of gel filtration and column chromatography. The presence of the monosaccharides has been established by the chemical analysis. The quantitative analysis of the alditol acetate derivatives of them showed the ratios of the monosaccharides analyzed by means of GC respectively. The concentrations of protein(280 nm) and carbohydrate(496 nm) were detected respectively. The information of the molecular weight from the pure polysaccharide was obtained by several standard Dextrans from the Sephadex chromatography.
