南大青叶薄层色谱法鉴别的完善与HPLC法含量测定的建立

目的:提高南大青叶的质量标准,对其质量进行更有效控制。方法:以靛蓝、靛玉红为对照品,通过薄层色谱法(TLC)对南大青叶进行定性分析;采用高效液相色谱法(HPLC)对靛蓝进行含量测定,以十八烷基硅烷键合硅胶为填充剂;以乙腈(A)-水(B)为流动相,梯度洗脱(0~10 min,5%→60%A;10~20 min,60%A);流速为1.0 mL·min^-1;检测波长为289 nm。结果:TLC斑点清晰,分离度高,方法耐用性好,能区分南大青叶及其混伪品广西马蓝;HPLC法含量测定结果显示,靛蓝在24.98~249.8 ng线性关系良好(r=0.9995,n=6),加样回收率为100.80%,RSD=1.8%,n=6。测定样品11批,结果靛蓝的含量范围为0.52%~0.92%。结论:所建立的薄层色谱鉴别方法专属性好,能有效鉴别该药材中靛蓝及靛玉红成分;HPLC方法分离度好、稳定可靠,可有效控制南大青叶药材的质量,为该药材的合理开发利用提供参考。     

Anti-tumor activity of noble indirubin derivatives in human solid tumor models In Vitro

Indirubin has been identified as a component of a traditional Chinese medicine, Danggui Longhui Wan, which is used for the treatment of chronic myelogenous leukemia. Indirubin inhibits cyclin-dependent kinases (CDKs) and induces cell cycle arrest and apoptosis in cancer cells. Many indirubin derivatives have been studied for their potential anti-solid tumor activity. We have synthesized and evaluated many indirubin derivatives. In order to compare and confirm the potential of our major derivatives as anti-solid tumor agents, we examined their anti-proliferative activity in monolayers, as well as in multicellular spheroids (MCS) cultures of human colorectal cancer cells, DLD-1 and HT-29. The MCS model is an in vitro solid tumor model that is increasingly used for the evaluation of anti-solid tumor activity. 5-nitro-indirubin-3′-oxime (4c) and 5′-bromo-5-nitro-indirubin-3′-oxime (4l), compared to 5-trimethylacetamido-indirubin-3′-oxime (11) and 5-diphenylacetamido-indirubin-3′-oxime (33) showed greater anti-proliferative effects in monolayers, but lower anti-proliferative effects in MCS. Overall, our data suggest that compounds 11 and 33 may exert a significant anti-solid tumor activity via a mechanism other than CDK inhibition, different from that of 4c and 4l. These compounds are worth further investigation with respect to their anti-solid tumor activity and their mechanism of action in various solid tumor models.     

复方贯众感冒滴鼻液的提取工艺研究

目的优选复方贯众感冒滴鼻液提取工艺的最佳条件。方法以靛玉红含量为考察指标,运用L9(34)正交试验设计,考察加水量、提取时间、提取次数等因素对提取工艺的影响。结果提取时间和提取次数对检测指标均有显著影响,加水倍数为不显著因数。实验结果表明,以加水量为药材的10倍量,煎煮时间1.5 h,提取2次为该制剂的最佳提取工艺。结论正交实验的最终确定方法简便、实用、经济,符合工业化生产的要求。     

板蓝根颗粒剂中靛玉红、腺苷含量测定方法学研究

目的：建立板蓝根颗粒剂中靛玉红、腺苷含量测定方法。方法：采用高效液相色谱法对板蓝根颗粒剂中靛玉红、腺苷进行了含量测定。结果：靛玉红在0.251～1.255μg范围内线性关系良好（r=0.9998）;平均回收率为98.61%,RSD为0.79%（n=5）;腺苷在38.12～190.60mg·L-1范围内线性关系良好,（r=0.9998）,平均回收率为102.80%,RSD为1.33%（n=5）。结论：高效液相色谱法对板蓝根颗粒剂中靛玉红、腺苷含量测定简便易行,准确可靠。     

复方感冒滴鼻液提取工艺的考察

目的考察复方感冒滴鼻液提取工艺的最佳条件。方法以靛玉红含量为考察指标,运用L9（3）^4正交试验设计．考察加水量、提取时间、提取次数等因素对提取工艺的影响。结果煎煮时间和煎煮次数对检测指标均有显著影响,加水倍数为不显著因数。结论以加水量为药材的10倍量、煎煮时间1．5h,煎煮2次为该制剂的最佳提取工艺。     

Opposite effects of lithium and valproic acid on trophic factor deprivation-induced glycogen synthase kinase-3 activation, c-Jun expression and neuronal cell death

Recent studies demonstrate that lithium and valproic acid (VPA), two commonly used mood-stabilizing drugs, have neuroprotective effects against a variety of insults. Inhibition of the proapoptotic enzyme, glycogen synthase kinase-3 (GSK-3), has been suggested to be the mechanism of action of neuroprotection for both drugs. In this study, we tested if lithium and VPA could protect cultured cerebellar granule neurons (CGNs) from GSK-3-mediated apoptosis induced by trophic factor withdrawal (serum/potassium deprivation). Both lithium and indirubin, a specific GSK-3 inhibitor, protected CGNs in a dose-dependent manner. In contrast, VPA did not provide any neuroprotection and even potentiated cell death. Immunoblot analysis revealed that lithium inhibited the trophic factor deprivation-induced activation of GSK-3 as well as the in vivo phosphorylation of the microtubule-associated protein Tau on Ser199, a specific target site for GSK-3. Under these same experimental conditions, however, VPA neither inhibited GSK-3 activation nor hindered GSK-3 mediated Tau phosphorylation. Furthermore, in accordance with their effects on neuronal survival, lithium prevented the induction of c-Jun expression in trophic factor-deprived CGNs, whereas VPA potentiated it. Collectively, these results show that VPA is not a universal inhibitor of neuronal GSK-3, and that instead of being neuroprotective, VPA can even exacerbate neuronal death under some conditions.     

HPLC测定青黛中的靛蓝和靛玉红

目的 建立青黛中靛蓝和靛玉红含量的测定方法.方法 采用HPLC法定量分析,色谱柱为DiamonsilTM C18(250 mm×4.6 mm,5 μm),流动相为甲醇-水(80:20),流速为1.0 ml·min-1,柱温为30℃;靛蓝的检测波长为287 nm,靛玉红的为292 nm.结果 靛蓝0.07～0.52 μg(r=0.9996)、靛玉红0.02～0.20 p,g(r=0.9999)与峰面积的线性关系良好(r=0.9996),其平均回收率分别为100.9%(RSD=1.42%,n=6)、101.3%(RSD=1.87%,n=6).结论 所建方法可用于青黛中靛蓝和靛玉红的含量测定,为完善青黛的质量标准提供了依据.     

靛玉红腭衍生物合成前体靛玉红色谱离析研究

目的:为合成新的高效、低毒靛玉红腭衍生物需对对原料靛玉红进行前处理,分离除去干扰性异构体杂质,建立合成靛玉红腭衍生物及其晶体培养的后继工作中有指导意义的常规色谱离析方法.方法:进行色谱筛选研究,用分离度Ks,选择因子a确定色谱离析系统.结果:S、t、Ks,、a、k显示乙酸乙酯/氯仿、乙酸乙酯/石油醚二元组分展开剂,G-CMC-Na、硅胶HCMC-Na的色谱离析可用于对原料的处理,以硅胶H-CMC-Na/乙酸乙酯/氯仿的色谱离析经重结晶可得到靛玉红暗红色细小针状晶体.结论:根据实验确定的Ks,a的色谱离析系统,可以实现合成反应中对原料靛玉红的前处理.     

靛玉红类化合物的抗肿瘤作用及构效关系的研究

Twenty-four indirubin derivatives or analogues have been subjected to experimental antitumor tests. Preliminary results revealed that 13 compounds showed marked inhibitory action against Walker 256 in rats with a tumor weight inhibition of 29⁶7%. Among them compounds 3,4,12,14 and 22 exhibited higher activity than indirubin, especially compounds 3 and 22. The structure-activity relationship of this series of indirubin derivatives and its analogues is discussed.