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《Immunopharmacology and immunotoxicology》2013,35(3):265-273
AbstractCinobufagin (CBG) is a cardiotoxic bufanolide steroid secreted by the skin and parotid venom glands of the Asiatic toad Bufo bufo gargarizans (called Chan-Su). Although CBG is known to exhibit anti-cancer activities, very little is known about its potential mechanism(s) of action. In this study, we investigated whether CBG mediates its effect through the modulation of the mitogen-activated protein kinases (MAPKs) signaling pathway in human multiple myeloma (MM) U266 cells. We found that CBG caused the significant activation of ERK, JNK and p38 MAPK in U266 cells. CBG showed much higher cytotoxicity against U266 cells as compared to peripheral blood mononuclear cells (PBMC). Induction of CBG increased reactive oxygen species (ROS) generation from mitochondria, which is associated with the induction of apoptosis as characterized by increased sub-G1 DNA contents of cell cycle, positive Annexin V binding, activation of caspase-3 and cleavage of PARP. Inhibition of ROS generation by N-acetyl-l-cysteine (NAC) significantly prevented CBG-induced ERK, JNK and p38 MAPK activation and apoptosis. CBG also down-regulated the expression of various downstream gene products that mediate cell proliferation, survival, angiogenesis and metastasis. Interestingly, ERK, JNK and p38MAPK pharmacological inhibitors blocked CBG-induced MAPKs activation and ERK inhibitor (PD98059) also prevented the CBG-induced caspase-3 activation and PARP cleavage in U266 cells. Taken together, these findings suggest that CBG can act as a potent anticancer agent against MM and possibly exerts its effects through the ROS-mediated activation of ERK, JNK and p38 MAPK leading to the activation of caspase-3 in U266 cells. 相似文献
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目的:建立用HPLC测定蒡芩慢咽滴丸中蟾酥内酯含量的方法。方法:采用十八烷基键合硅胶色谱柱(250mm×4.6mm,5μm),以乙腈-0.5%磷酸二氢钾溶液(用磷酸调pH值为3.2)(32∶68)为流动相,检测波长为296nm,柱温40℃。结果:酯蟾毒配基及华蟾酥毒基的线性范围分别为0.2~1.0μg(r=0.9997)及0.224~1.12μg(r=0.9996),平均回收率(n=6)分别为99.91%(RSD=0.7960%)和99.84%(RSD=1.0023%)。结论:该方法简单可靠。 相似文献
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液相色谱法测定强力救心滴丸中蟾酥的华蟾酥毒基和酯蟾毒配基含量 总被引:2,自引:0,他引:2
目的:测定强力救心滴丸中蟾酥的主要成分华蟾酥毒基和酯蟾毒配基含量。方法:样品经甲醇超声处理提取,采用高效液相色谱法测定,色谱柱为 Alltima C_(18)色谱柱(4.6mm×250mm,5μm);柱温:40℃;流速:1mL·min~(-1);流动相:乙腈-0.5%磷酸二氢钾(pH=3.2)(50:50);检测波长:296nm。结果:华蟾酥毒基在0.25-1.51μg 范围内、酯蟾毒配基在0.26-1.55μg 范围内具有良好线性关系。华蟾酥毒基平均加样回收率为99.6%,RSD=2.1%(n=5);酯蟾毒配基平均加样回收率为98.2%,RSD=1.4%(n=5)。结论:该法准确、可靠,可用于强力救心滴丸中蟾酥的酯蟾毒配基和华蟾酥毒基含量测定。 相似文献
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目的 建立检测大鼠血浆中3种蟾蜍甾烯类化合物蟾毒灵、华蟾酥毒基及酯蟾毒配基的HPLC法,并用于蟾酥在大鼠体内的药动学研究。方法 分别于大鼠尾iv给予蟾酥提取物0.8 mg/kg后2、5、10、15、20、30、45、60、90 min,眼眶取血,采用乙腈沉淀蛋白与液液萃取相结合方法进行血浆样品预处理,以HPLC法测定大鼠血浆中蟾毒灵、华蟾酥毒基及酯蟾毒配基的质量浓度,以Kinetica软件拟合药动学参数。结果 蟾毒灵、华蟾酥毒基及酯蟾毒配基均得到很好的分离,重现性、精密度、线性关系良好,达到体内分析要求;经非房室模型拟合,得到蟾毒灵、华蟾酥毒基及酯蟾毒配基在大鼠体内主要药动学参数;iv给药30 min时,血药浓度均降至Cmax的1/5以下。结论 建立的蟾毒灵、华蟾酥毒基及酯蟾毒配基血药浓度测定方法操作简单、结果准确可靠;蟾蜍甾烯类成分在大鼠体内代谢较为迅速,所得数据可为蟾酥提取物的药动学研究提供参考。 相似文献
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蟾酥通痹软膏的质量标准研究 总被引:3,自引:0,他引:3
目的:建立蟾酥通痹软膏(蟾酥、冰片、防己等)的质量标准。方法:采用TLC法对处方中的防己等进行定性鉴别;用HPLC法测定了华蟾酥毒基和脂蟾毒配基的含量。结果:在TLC图谱中可检出防己等药材的特征斑点;华蟾酥毒基在1.15μg~5.75μg之间线性关系良好,r=0.999 7,精密度实验RSD为1.05%,平均回收率为97.95%,(RSD=1.63%,n=5);脂蟾毒配基在1.05μg~5.25μg之间线性关系良好,r=0.999 6,精密度实验RSD为1.02%,平均回收率为97.45%,(RSD=0.61%,n=5)。结论:所采用方法准确、可靠,可行性及重复性良好,能有效控制该制剂的质量。 相似文献
7.
目的用HPLC法测定蟾皮中蟾蜍灵、脂蟾毒配基、华蟾蜍精等成分的含量。方法样品经乙醇室温浸渍提取,采用HPLC法测定。色谱柱为DiamonsilTMC18(250 mm×4.6 mm,5μm),流动相为甲醇-乙腈-水(体积比为4∶1∶5),流速为1.0 mL.min-1,检测波长为296 nm,柱温为40℃。结果蟾蜍灵、脂蟾毒配基和华蟾蜍精分别在0.0848~0.5088、0.0884~0.5304和0.2000~1.2000μg内与色谱峰面积线性关系良好,相关系数r分别为0.9999、0.9992和1.0000。蟾蜍灵、脂蟾毒配基和华蟾蜍精的平均回收率分别为99.3%、99.6%和99.0%,RSD分别为0.5%、0.2%和0.6%(n=9)。结论HPLC法可用于蟾皮中蟾蜍灵、脂蟾毒配基和华蟾蜍精的含量测定。 相似文献
8.
目的建立消炎解毒软膏中华蟾酥毒基和脂蟾毒配基含量测定方法。方法采用HPLC法测定消炎解毒软膏中华蟾酥毒基和脂蟾毒配基的含量,色谱条件为Alltima C18:色谱柱(4.6mm×250mm,5μm);流动相:水-乙腈(50∶50),流速:1mL·min-1;检测波长:296nm;柱温:35℃。结果华蟾酥毒基的进样量在0.427~17.080μg峰面积积分值与进样量有良好的线性关系(r=0.9999),平均回收率为98.3%,RSD为2.0%(n=6);脂蟾毒配基的进样量在0.262~10.480μg峰面积积分值与进样量有良好的线性关系(r=0.9999);平均回收率为98.9%,RSD为1.8%(n=6)。结论本方法简便、快速准确、灵敏度高、重现性好,可作为该制剂的含量测定方法。 相似文献
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Xiao-Liang Wang Guang-Hou Zhao Jin Zhang Qi-Yun Shi Wei-Xiao Guo Xiu-Li Tian 《Journal of Asian natural products research》2013,15(5):383-392
The objective of this study was to evaluate the immunomodulatory effects of cinobufagin (CBG) isolated from Chan Su (Venenum Bufonis) in vitro. In this paper, our results show that CBG signi?cantly stimulated cell proliferation of splenocytes and peritoneal macrophages (PMΦ) and markedly enhanced the phagocytic activation of PMΦ. CBG also significantly increased CD4+CD8+ double-positive T-cell populations and the percentage of S-phase cells of splenic lymphocytes. The levels of several Th1 cytokines, including interferon-γ and tumor necrosis factor-α, are signi?cantly increased after CBG treatment, whereas the levels of the Th2 cytokine interleukin-4 and interleukin-10 are signi?cantly decreased. As a result, the ratio of Th1/Th2 also increased. Taken together, these results indicated that CBG had potential immune system regulatory effects and suggested that this compound could be developed as a novel immunotherapeutic agent to treat immune-mediated diseases such as cancer. 相似文献