Effect of yeast elicitor and salicylic acid on the fluctuation of phytohormone contents in Ti-transformed Salvia miltiorrhiza cell cultures

When Ti transformed Salvia miltiorrhiza cells werecultured in a MS-NH₄ medium (MS without ammonium nitrate, containing30 g/L sucrose) at 25 °C in darkness for 18d, the total tanshinone (cryptotanshinone and tashinone IIA)contents in cultures were 12.23 mg/L and 15.07 mg/Lfor yeast elicitor (4 g/L), and yeast elicitor plus 200mol/L salicylic acid (SA) treated cultures, respectively,whereas only trace amounts of tanshinone were detected in the control or SAtreated cells. To explore the hormonal background concerning these phenomena,endogenous phytohormones were determined using ELISA kits. We found that ABA andiPAs contents in yeast elicitor plus SA treated cell cultures were increased 2.8to 9.8-fold and 3.6 to 5.8-fold respectively, while contents of GA₁and IAA were decreased by 13.2%–56.9% and 34.8%–74.6% respectively.This suggests that higher levels of ABA and iPAs combined with lower levels ofGA₁ and IAA inhibit the growth of cells, then probably stimulate thetanshinone production.     

Induction of CYP1A by a diterpene quinone tanshinone IIA isolated from a medicinal herb Salvia miltiorrhiza in C57BL/6J but not in DBA/2J mice

Effects of tanshinone IIA, an active diterpene quinone of the herbal medicine Salvia miltiorrhiza (Danshen), on cytochrome P450 (CYP), UDP-glucuronosyl transferase (UGT), and glutathione S-transferase (GST) were studied in the arylhydrocarbon (Ah)-responsive C57BL/6J (B6) and nonresponsive DBA/2J (D2) mice. Oral treatment of tanshinone IIA caused a dose-dependent increase of liver microsomal 7-methoxyresorufin O-demethylation (MROD) activity in B6 but not in D2 mice. In B6 mice, tanshinone IIA increased hepatic benzo(a)pyrene hydroxylation (AHH), 7-ethoxyresorufin O-deethylation, MROD, and 7-ethoxycoumarin O-deethylation activities. The levels of Cyp1A2 protein and mRNA were elevated. On the contrary, in D2 mice, tanshinone IIA decreased hepatic AHH and nifedipine oxidation activities and the CYP3A protein level without affecting other activities determined. Cyp1A2 protein and mRNA levels were not affected by tanshinone IIA in D2 mice. Tanshinone IIA had no effects on UGT and GST activities in both B6 and D2 mice. These results demonstrated that induction of CYP1A2 by tanshinone IIA depended on the Ah-responsiveness and occurred at pre-translational level.     

Tanshinone II-A inhibits low density lipoprotein oxidation in vitro

Tanshinone II-A (TSII-A) is a major component of Salvia miltorrhiza Bunge which has long been used for preventing and ameliorating anginal pain in China. However the effect of TSII-A on low density lipoprotein (LDL) oxidation has not been studied. The present study was performed to investigate the effects of TSII-A on LDL oxidation using four oxidizing systems, including copper-, peroxyl radical- and peroxynitriteinitiated and macrophage-mediated LDL oxidation. LDL oxidation was measured in terms of formation of thiobarbituric acid-reactive substances (TBARS), relative electrophoretic mobility (REM) on agarose gel and lag time. In all four systems, TSII-A has apparent antioxidative effects against LDL oxidation, as evidenced by its dose-dependent inhibition of TBARS formation, prolongation of lag time and suppression of increased REM.

Regarding the mechanism underlying its antioxidative effect, TSII-A neither scavenged superoxide nor peroxynitrite. It also did not chelate copper. But it has mild peroxyl radical scavenging activity. The direct binding to LDL particles and conformational change of LDL structure by TSII-A were suggested, because it increased negative charge of LDL which was shown by increased REM on agarose gel. In conclusion, TSII-A is an effective antioxidant against LDL oxidation in vitro. The underlying mechanism appears to be related to its peroxyl radical scavenging and LDL binding activity.     

丹参活性成分的现代中药药理研究进展

对传统中药丹参中丹参酮、丹酚酸、丹参单体IH764-3三种主要活性成分及其近年来的中药药理学研究进展进行综述,为进一步开发利用这一传统中药提供理论依据。     

PEG-amino acid-przewaquinone a conjugations: Synthesis,physicochemical properties and protective effect in a rat model of brain ischemia-reperfusion

A total of 21 PEG-przewaquinone A conjugations with high drug loading ability, good water solubility and in vivo slow-release quality were obtained by conjugating przewaquinone A with PEG through amino acids and tripeptides spacers respectively. Notably, compound 3a can obviously reduce the brain ischemia-reperfusion damage dose-dependently in a rat model, which indicated the efficacy of our PEG prodrug strategy.     

Mutation breeding of Emericella foeniculicola TR21 for improved production of tanshinone IIA

TR21, an original tanshinone IIA-producing endophytic fungus from the root of Salvia miltiorrhiza Bunge, produces low levels of tanshinone IIA. Three techniques were used to rapidly improve tanshinone IIA production: TR21 mutation by ultraviolet radiation (UV), TR21 mutation by sodium nitrite (NaNO₂) and TR21 mutation by a combination of both UV and NaNO₂. An improved mutant, labeled as NU152, was obtained from the combination of UV and NaNO₂. The content of tanshinone IIA produced by NU152 held a relative high value of 51.44 ± 0.22 μg per g, and the NU152 produced a more than 1.46-fold increase in tanshinone IIA yield, compared with the wild type TR21 (P < 0.05). The fungus NU152 showed a possible way for the sustainable production of tanshinone IIA.     

丹参酮ⅡA对正常和氧化损伤内皮细胞的保护作用研究

采用过氧化氢诱导建立的内皮细胞氧化损伤模型,探讨了丹参酮ⅡA对氧化损伤的HUVECs的保护作用.通过内皮细胞LDH、SOD、NOS、GSH-Px的活性和MDA、NO含量的变化趋势,MTT和流式细胞术反映的细胞存活率的变化,研究了丹参酮ⅡA对正常和氧化损伤HUVECs的作用.结果表明丹参酮ⅡA在10～100μmol/L浓度之间时,无明显细胞毒性.浓度在20 μmol/L以上时,丹参酮ⅡA能显著增强HUVECs抵御氧化损伤的能力;同时降低了LDH的泄漏,阻止了MDA等过氧化物的生成,提高了NOS、NO、SOD和GSH-PX分泌量,显著减少了细胞早期凋亡率和坏死率.表明丹参酮ⅡA可以保护HUVECs免受氧化损伤,防止动脉粥样硬化斑块的形成.     

丹参酮IIA衍生物的合成

目的:丹参酮IIA是中药丹参的脂溶性成分,具有抗肿瘤、抗氧化、抗心脑血管疾病等多种生理活性。本文拟对其进行结构改造以获得活性更好的丹参酮IIA衍生物。方法:首先,以丹参酮IIA为原料,通过Vilsmeier反应在其16-位引入醛基,再与醋酸胺进行还原胺化反应,以较高收率得到16-位胺甲基取代的丹参酮IIA衍生物。接着,对其氨基进行修饰,得到10个不同N-取代的丹参酮IIA衍生物。同时考察反应温度、反应溶剂和反应时间等条件对还原胺化反应的影响,确定最佳反应条件。结果:通过1H-NMR、13C-NMR以及LC-MS对所有产物结构进行了确认。还原胺化反应的最佳反应条件为:以1,2-二氯乙烷为溶剂,温度保持40℃,反应时间为2h。结论:反应步骤简单、条件温和、产率较高,是合成16-位取代的丹参酮IIA衍生物的理想方法。     

丹参酮的药理作用研究进展

丹参酮在中药丹参中的含量较高,有较强的生理活性,是丹参主要有效成分之一。丹参酮的药理作用极其广泛,本文就丹参酮的抗肿瘤、抗动脉粥样硬化、抗心律失常、缩小心肌梗死面积、降低心肌耗氧量、逆转左心室肥厚、减轻缺血再灌注损伤、保肝脏及抗肝纤维化、保护心肌、改善微循环、抗菌消炎等作用进行综述,以便促进中药丹参在临床上广泛的应用。     

Tanshinone IIA, an isolated compound from Salvia miltiorrhiza Bunge, induces apoptosis in HeLa cells through mitotic arrest

AIMS: Tanshinone IIA (Tan IIA) is a compound isolated from Salvia miltiorrhiza Bunge (Danshen). The aim of this study is to investigate the mechanisms of its anti-cancer effect. MAIN METHODS: To clearly delineate the cell cycle-dependent effects of Tan IIA, we used either synchronized cells or single living cell analysis to conduct our studies. Subcellular fractionation, Western blot analysis, immuno-fluorescence staining and FACS analysis were also employed in our study. KEY FINDINGS: We found that Tan IIA could arrest cancer cells in mitosis by disrupting the mitotic spindle and subsequently triggered cells to enter apoptosis through the mitochondria-dependent apoptotic pathway. Thus, Tan IIA could selectively kill mitotic cells over interphase cells. In comparison with other existing anti-cancer drugs that cause mitotic arrest by interfering with the microtubule structure (such as vincristine or taxol), Tan IIA destroyed only the mitotic spindle during the M phase but not the microtubule structure in interphase cells. Furthermore, Tan IIA could trigger the mitotic arrested cells to enter apoptosis faster than vincristine or taxol. SIGNIFICANCE: Since Tan IIA can selectively induce cancer cells to enter apoptosis through mitotic arrest, it has the potential to be developed into an anti-cancer drug.