超高压水射流对丹参脂溶性成分丹参酮ⅡA的影响研究

[目的]研究超高压水射流对丹参脂溶性成分丹参酮ⅡA的影响。[方法]用150~350 MPa不同压力的水射流处理丹参醇提液,采用薄层色谱法和高效液相色谱法定性鉴别和测定丹参的化学成分种类及含量变化。[结果]不同压力水射流处理丹参提取液后,丹参的薄层图谱中斑点个数、大小及Rf值无明显差别;丹参的液相图谱中峰个数及形状亦无明显差别,但经150~300 MPa压力的水射流处理后,丹参酮ⅡA损失率达50%,经350 MPa压力的水射流处理后,丹参酮ⅡA的损失率将近80%。[结论]该研究为超高压水射流对中药有效成分的影响研究提供基础试验数据,为研究中药新的加工处理方法进行了有益的探索。     

丹参酮I抑制小鼠B16黑色素瘤细胞研究

将丹参酮I（含量为90%）配制成不同浓度作用于体外培养的B16细胞,对B16细胞用MTT法检测药物对细胞生长抑制率,收集并采用PI染色,观察细胞的形态,通过流式细胞术（FCM）检测细胞周期及凋亡率。结果表明：不同质量浓度的（5、4、3μg/mL）丹参酮I对B16细胞的抑制作用均有明显的剂量和时间效应;丹参酮I可以使B16细胞周期的构成发生明显的变化;丹参酮I对B16细胞的生长有明显的抑制作用,作用机制之一是使S期细胞的比例降低,促使细胞凋亡。     

丹参主要农艺性状与丹参酮ⅡA、丹酚酸B的相关性及通径分析

采用田间小区试验,对27个丹参品系的形态学性状与丹参酮ⅡA、丹酚酸B的含量进行相关性及通径分析。结果表明,丹参酮ⅡA含量与花轮数、根条数呈显著正相关。丹酚酸B含量与分枝数呈显著正相关,与花枝数呈极显著正相关。可见,花轮数、根条数和分枝数可作为高丹参酮ⅡA和丹酚酸B含量丹参品种选育的重要指标。     

中药有效成分丹参酮IIA、三七总苷与阿司匹林抗结直肠癌机制的研究现状及展望

结直肠癌是消化系统最常见的恶性肿瘤之一,并且其发病率呈逐渐升高的趋势。大量研究发现阿司匹林具有降低结直肠癌发生率的可能,而最新研究发现在具有活血化瘀的中药丹参及三七的提取物丹参酮ⅡA及三七总苷也发现了同样的疗效,因此本文就阿司匹林、丹参酮ⅡA及三七总苷抗结直肠癌的机制研究作一综述,以期为其下阶段研究和开发提供参考。     

丹参饮片产地加工工艺与质量研究

[目的]研究不同加工方法对丹参饮片质量的影响,为改良丹参饮片产地加工工艺提供参考。[方法]采用干切、浸润、水蒸气闷润3种方法加工丹参饮片,通过饮片品相认定并结合有效成分含量测定综合评价丹参饮片质量。[结果]丹参干切片切制困难,表面色泽差,呈砖红色,常出现开裂、翘边,片形不一致且成片率低;浸润所得丹参饮片表面颜色加深,切面平整、无裂缝、呈暗红色,芯部呈紫黑色,片形不美观,丹酚酸B、丹参酮ⅡA含量均显著降低;水蒸气闷润所得丹参饮片表面颜色鲜亮、呈紫红色,切面平整、光滑、无裂痕、呈淡黄色,质硬,参味浓,片形美观,成片率高,加工省时省力,且丹酚酸B与丹参酮ⅡA含量无显著变化。[结论]从饮片品相、各活性成分含量、生产成本等方面综合考虑,丹参饮片的产地加工方法应以水蒸气闷润软化药材为宜。     

Tanshinone ⅡA attenuates doxorubicin-induced injury in H9c2 cardiomyocytes via AMPK-mediated autophagy

AIM: To investigate the effect of tanshinone ⅡA on doxorubicin-induced rat H9c2 cardiomyocyte injury. METHODS: The H9c2 cardiomyocytes were treated with tanshinone ⅡA and/or doxorubicin with or without AMPK inhibitor dorsomorphin. The cell viability was measured by CCK-8 assay, the LDH release was examined for evaluating the cell injury, autophagy was analyzed by immunofluorescence staining, and AMPK activation was determined by Western blot. RESULTS: Compared with control group, the viability of H9c2 cells was decreased, the release of LDH was increased, the autophagy degree was increased, and AMPK activation was inhibited after treatment with doxorubicin (P<0.05). Compared with doxorubicin group, the treatment with tanshinone ⅡA restored the cell viability, reduced the release of LDH, further increased autophagy degree, and activated AMPK in the H9c2 cells (P<0.05). AMPK inhibitor dorsomorphin attenuated the abilities of tanshinone ⅡA to restore the cell viability, reduce the release of LDH, and increase autophagy degree in the H9c2 cells (P<0.05). CONCLUSION: Tanshinone ⅡA attenuates doxorubicin-induced injury in H9c2 cardiomyocytes, and its mechanism might be related to AMPK-mediated autophagy, which provides experimental and theoretical basis for the clinical application of tanshinone ⅡA in the prevention and treatment of doxorubicin-induced cardiomyocyte injury.     

Tanshinone IIA inhibits doxorubicin resistance in gastric cancer cells

AIM: To investigate the inhibitory effect and the specific mechanism of tanshinone IIA on doxorubicin (DOX)-resistant gastric cancer cells. METHODS: The sensitivity of gastric cancer cells lines to DOX was determined by MTT assay. DOX-resistant gastric cancer cell lines were established by step selection with increasing concentrations of DOX. The cell cycle arrest, apoptosis and autophagy related-markers were analyzed by flow cytometry and Western blot. The expression of P-glycoprotein (P-gp), breast cancer resistance protein (BCRP) and multi-drug resistance-associated protein 1 (MRP-1) was determined by RT-qPCR and Western blot. RESULTS: DOX-sensitive cell lines SNU-719 and SNU-601 as well as the cell lines relatively resistant to DOX including SNU-638, SNU-668, SNU-216 and SNU-620 were identified according to the IC₅₀ values of DOX for different cell lines. Two DOX-resistant cell lines SNU-719R and SNU-601R were also established. Tanshinone IIA inhibited the expression of MRP-1 in DOX-resistant cell lines. Compared with DOX treatment alone group, combined treatment of DOX and tanshinone IIA in cancer cells decreased the G₂/M phase cell number, increased the protein expression of p21, decreased the protein expressions of cyclin B1 and cyclin-dependent kinase 1 (CDK1) in the SNU-719 R cells and SNU-620 cells. In addition, compared with DOX treatment alone group, combined treatment of DOX and tanshinone IIA in the cancer cells increased the protein expressions of p53, Bax and LC3B-II, decreased the protein expression of Bcl-2 and p62 (P<0.05). CONCLUSION: Tanshinone IIA is an effective drug in the inhibition of DOX resistance in gastric cancer.     

HPLC法测定丹参保健茶中3种丹参酮含量

[目的]用HPLC测定丹参保健茶中隐丹参酮、丹参酮Ⅰ和丹参酮ⅡA的含量。[方法]色谱条件为：色谱柱为Insertsil ODS-SP（4.6mm×250 mm,5μm）;流动相为乙腈-水（V∶V=62∶38）,等度洗脱;检测波长为270 nm;流速为1.0 ml/min;进样量为15μl,柱温30℃。[结果]3种丹参酮成分得到较好地分离,丹参保健茶中隐丹参酮、丹参酮Ⅰ、丹参酮ⅡA的平均回收率分别为98.2%、98.7%、98.4%（RSD值分别为1.20%、1.96%和1.74%,n=3）;样品中3种丹参酮含量分别为1.19、0.79和2.25 mg/g。[结论]该方法稳定可靠、简便快速,可用于丹参保健茶的质量控制。     

丹参药渣中提取丹参酮的研究(摘要)(英文)

[目的]水浸丹参和三七后药渣中提取丹参酮并确定丹参酮成分。[方法]采用有机溶剂法提取丹参药渣中丹参酮,7种有机溶剂分别为甲醇、正己烷、石油醚、乙酸乙酯、丙酮、乙醚、二氯甲烷、三氯甲烷。以薄层层析法检测确定最佳提取溶剂。在薄层层析板上,用毛细管吸取标准品及样品,点样于薄层层析板上,每次点样依照样品浓度确定点样次数,每次点样均须用电吹风吹干。丹参酮展开剂的配方为:石油醚∶乙酸乙酯∶冰乙酸=8∶3∶1(V∶V∶V),直接观察;三七皂苷展开剂为V(氯仿)∶V(甲醇)∶V(水)=14∶6∶1,用10%H2SO4加热显色。通过高效液相色谱法确定丹参药渣中丹参酮成分。高效液相色谱法(HPLC):高效液相色谱仪为美国Waters2695-2996DAD;色谱柱为HypersilODS25μm(4.6mm×150.0mm)fromElite;进样量,10μl;体积流量,1.0ml/min。洗脱液A为水,B为甲醇。洗脱条件,初始时,A为40%,B为60%;5min时,A为40%,B为60%;20min时,A为20%,B为80%;30min时,A为20%,B为80%。[结果]乙醚为丹参药渣中提取丹参酮的最佳溶剂。水浸丹参和三七后干药渣,先经乙醇提取得到脂溶性提取物,再以乙醚为溶剂进行索氏提取,丹参酮混合物得率为2.17%;高效液相色谱法检测,丹参酮混合物中丹参酮ⅡA、次甲丹参醌、隐丹参酮、丹参酮I含量分别为3.62%、1.02%、2.56%、2.75%。[结论]丹参药渣中丹参酮成分与药材丹参基本一致,丹参药渣可以作为一种丹参酮资源,具有开发利用的价值。