新二萜醌去氢丹参新酮

从鼠尾草属植物红根草(Salvia prionitis Hance)的根中分到四个二萜醌类化合物,经光谱分析推定为去氢丹参新酮(Ⅰ),丹参新酮(Ⅱ),丹参新醌乙(Ⅲ)和丹参新醌丁(Ⅳ)。Ⅰ为新化合物,所有化合物均系从该植物中首次分到。     

人参叶微量新成分的研究

从人参(Panax ginseng C.A.Meyer)叶中分离出14种单体化合物。其中4种微量成分用IR、MS、¹HNMR、13CNMR及化学方法等分别鉴定为20(R)-原人参三醇(Ⅰ)、胡萝卜甙(Ⅱ)、3β,12β-二羟基-20(22),24-达玛二烯—3—0—β—D—葡萄吡喃糖甙(Ⅲ)及20(R)-原人参二醇—3—0—β—D—葡萄吡喃糖甙(Ⅳ)。其中Ⅲ及Ⅳ系新化合物,分别命名为人参皂甙-Rh₃(ginsenoside-Rh₃)及20(R)-人参皂甙-Rh₃[20(R)-ginsenoside-Rh₂]。     

丹参有效成分的研究——Ⅴ.紫丹参甲素和乙素的分离和化学结构

从云南产紫丹参(Salvia przewalskii Maxim.var.mandarinorum Stib.)中分离到两种新的二萜醌类化合物,取名为紫丹参甲素(C₁₉H₁₈O₄,熔点173～5℃)和紫丹参乙素(C₁₄H₁₂O(4),熔点242～3℃),经物理和化学方法的研究推定其结构式分别为(Ⅰ)和(Ⅱb)。紫丹参甲素有抗动物肿瘤活性,且其抑菌作用比隐丹参酮强。试探了紫丹参乙素质谱中M-16裂片的裂解由来。     

中国辽宁栽培西洋参化学成分的研究

中国辽宁栽培西洋参(Panax quinquefolius Linn)的总皂甙用低压硅胶柱和反相Rp18Labar柱层析分离得到18种化合物,用IR,MS(FD-MS,FAB-MS),¹³C-NMR及化学方法鉴定了16种化合物的化学结构;分别为棕榈酸(1),齐墩果酸(2),胡萝卜甙(daucosterin 3),人参皂甙-Rh₁(4),—Rg₃(5),—Rg₂(6),—Rg₁(7),—Rf(8),—Re(9),—Rd(10),—Rb₂(11),—Rb₁(12)、—R₀(13),蔗糖(14),人参三糖(15)及一种新皂甙(16),结构为:20(s)原人参二醇-3-[-O-β-D-吡喃糖基(1→2)β-D-葡萄吡喃糖基(1→2)β-D-葡萄吡喃糖基],20-[-O-β-D-葡萄吡喃糖基(1→6)β-D-葡萄吡喃糖甙,命名为人参皂甙-RAO(ginsenoside-RA₀)。化合物(4)和(5)系首次从西洋参中分离出的已知皂甙。     

6α-甲基-17α-羟基-黄体酮和6α-甲基-17α-羟基-11去氧-皮质酮-21-乙酸酯的微生物羟化

6α-甲基-11-去氧-17α-羟基-皮质酮-21-乙酸酯(Ⅱ)经短刺克宁汉霉微生物转化得6β-羟基化合物(Ⅵ_a)及6β,11β-羟基化合物(Ⅶ_a)。6α-甲基-17α-羟基-黄体酮(Ⅰ)经短刺克宁汉霉菌转化亦得6β-羟基化合物(Ⅵ_b)及6β,11β-羟基化合物(Ⅶ_b)。化合物(Ⅱ)如用梨头霉转化则得11α-羟化物(Ⅲ)。Ⅶ_a、Ⅶ_b、Ⅷ_a、Ⅷ_b及Ⅺ_b的结构是通过核磁共振谱和质谱证明的。     

血吸虫病化学治疗的研究——ⅩⅩⅩⅦ.β-(5-硝基-4-溴-2-呋喃)-及β-(4,5-二溴-2-呋喃)丙烯酰胺及其酯类的合成

本文报道β-(4,5-二溴-2-呋喃)-及β-(5-硝基-4-溴-2-呋喃)丙烯酰胺及其酯类衍生物26个的合成。动物筛选结果表明;化合物Ⅲ₆,Ⅲ₈和Ⅲ₁₃对感染日本血吸虫小白鼠有明显的治疗作用。化合物Ⅱ₆有较明显的预防作用。     

蔓性千斤拔化学成分的研究

从豆科植物蔓性千斤拔[Flemingia philippinensis(Merr.et Rolfe)Li]的根中分得六个化合物和一个烷酸的混合物(Ⅰ～Ⅶ)。经理化常数测定和光谱分析证明,化合物Ⅰ和Ⅱ为新化合物,分别命名为蔓性千斤拔素C(Ⅰ)和蔓性千斤拔素D(Ⅱ),其余为已知物,分别鉴定为5,7,3′,4′-四羟基-6,8-双异戊烯基异黄酮(Ⅲ),flemichin D(Ⅳ),n C₂₂～C₃₀烷酸(Ⅴ),β-谷甾醇(Ⅵ)和羽扇豆醇(Ⅶ)。化合物Ⅲ和Ⅳ对P₃₈₈白血病细胞具有显著的抑制作用。     

华北白前中的C21甾体类化合物

从华北白前(Cynanchum hancockianum)干燥根的乙醇提取物中分离鉴定了6个C₂₁甾体化合物,其中白前甙元C(glaucogenin C,Ⅰ),白薇甙A(cynatratoside A,Ⅱ),白前甙元A(glaucogenin A,Ⅲ),脱水何拉得甙元(anhydrohirundigenin,Ⅳ)为已知化合物;华北白前甙元B(hancogenin B,Ⅴ),华北白前甙A(hancoside A,Ⅵ)为新化合物。提出了变型C₂₁甾体甙元质谱的裂解规律。药理实验发现化合物Ⅱ具有一定的抗肿瘤活性,化合物Ⅵ具有一定的抗内毒素作用。     

傣药小灯台中的吲哚生物碱

从傣药小灯台(Winchia catophylla A. DC.)中分到4个吲哚生物碱,经理化常数测定,光谱分析和化学转化,分别鉴定为echitamine chloride(Ⅰ),echitamidine(Ⅱ),N_B-demethyl-echitamine(Ⅲ)和22-O-acetyl-N_b--demethyl-echitamine(Ⅳ),其中Ⅳ为新的吲哚生物碱。     

基于网络药理学和分子对接技术探讨丹参治疗缺血性脑卒中的作用机制

目的 利用网络药理学、分子对接技术，筛选并分析丹参治疗缺血性脑卒中（CIS）的关键化合物、作用靶点、生物学功能及信号通路，阐述其可能的作用机制。方法 使用TCMSP数据库检索丹参的活性成分及作用靶点，在GeneCards、NCBI和OMIM数据库中获取CIS的靶点，并将药物和疾病交集后的靶点输入STRING数据库，构建蛋白质相互作用网络，利用Cytoscape 3.7.2软件构建药物-化合物-作用靶点-疾病网络。通过Bioconductor进行GO功能富集和KEGG通路分析。采用AutoDock Tools 1.5.6、AutoDock vina 1.1.2将得到的关键活性成分和核心靶点进行分子对接，运用Pymol和Ligplot软件对结果进行可视化。结果 从丹参中共筛选出65个活性成分和108个对应靶点，以及CIS交集后获得靶点87个，其中度值大于50的靶点有6个，包括AKT1、IL6、FOS、VEGFA、MAPK1、EGFR，即本研究的核心靶点。GO功能富集分析得到GO条目124个（P<0.05），KEGG通路富集分析筛出134条信号通路（P<0.05），以PI3K-AKT信号通路所占靶点数目最多。构建的药物-化合物-作用靶点-疾病网络显示，木犀草素、丹参酮Ⅱ_A等活性化合物在整个网络中发挥着关键作用；分子对接结果显示，木犀草素、丹参酮Ⅱ_A与6个核心靶蛋白均具有较好的亲和力（结合能远小于-5 kJ/mol）。结论 丹参的主要活性化合物，包括木犀草素、丹参酮Ⅱ_A等，可作用于AKT1、IL6、FOS、VEGFA、MAPK1、EGFR等核心靶点，共同参与调节PI3K-AKT等多条信号通路，发挥抑制细胞凋亡、抗炎、神经保护等作用，可能是丹参治疗CIS的潜在机制。     

甘西鼠尾根化学成分研究甘西鼠尾根化学成分研究

目的研究甘西鼠尾(Salvia przewalskii Maxim)的化学成分。方法应用硅胶柱色谱法对甘西鼠尾根中化学成分进行分离,根据理化常数测定和光谱(IR,FAB-MS,EI-MS,¹HNMR,¹³CNMR,¹H-¹H COSY,¹H-¹³C COSY,HMBC,NOESY)分析鉴定化合物的结构。结果获得并鉴定了8个化合物结构:丹参酮II-A(I),隐丹参酮(II),紫丹参甲素(III),柳杉酚(IV),熊果酸(V),2α,3α-二羟基乌苏-12-烯-28-酸(VI),齐墩果酸(VII),甘西鼠尾新酮A(VIII)。结论VIII为新化合物,II,IV,V,VI和VII均为首次从该植物中分得。     

Inhibitory effects of tanshinone Ⅱ-A on invasion and metastasis of human colon carcinoma cells

Aim:

To investigate the effects and possible mechanisms of tanshinone II-A, an alcohol extract of the root of Salvia miltiorrhiza Bunge, on tumor invasion and metastasis of human colon carcinoma (CRC) cells.

Methods:

The effects of tanshinone II-A on invasion and metastasis of CRC cell lines HT29 and SW480 were evaluated by in vitro and in vivo assays. Western blotting was used to investigate possible molecular mechanisms of tanshinone II-A anti-cancer actions.

Results:

Tanshinone II-A inhibited migration and invasion of CRC cells in a dose-dependent manner. The inhibitory effect also depended on time, with the most significant effects observed at 72 h. Tanshinone II-A also significantly inhibited in vivo metastasis of colon carcinoma SW480 cells. It inhibited in vitro and in vivo invasion and metastasis of CRC cells by reducing levels of urokinase plasminogen activator (uPA) and matrix metalloproteinases (MMP)-2 and MMP-9, and by increasing levels of tissue inhibitor of matrix metalloproteinase protein (TIMP)-1 and TIMP-2. Tanshinone II-A was also shown to suppress the nuclear factor-kappaB (NF-κB) signal.

Conclusion:

Tanshinone II-A inhibited in vitro and in vivo invasion and metastasis of CRC cells. The effect resulted from changes in the levels of uPA, MMP-2, MMP-9, TIMP-1 and TIMP-2, and apparent inhibition of the NF-κB signal transduction pathway.     

丹参酮II-A通过调控TLR4/IκBα/NFκB信号通路抑制LPS诱导的细胞炎症

目的建立脂多糖(LPS)诱导的小鼠单核巨噬细胞(RAW264.7)炎症模型,探究丹参酮II-A(Tan IIA)的抗炎活性及其机制。方法CCK-8法测定Tan IIA对细胞活力的影响;迁移小室测定Tan IIA对LPS诱导细胞迁移能力作用;ELISA法测定细胞上清液中小鼠肿瘤坏死因子α(tumor necrosis factoralpha,TNF-α)、白介素6(interleukin 6,IL-6)、IL^-1β、单核细胞趋化蛋白-1(monocyte chemoattractant protein,MCP-1)的含量;Western blot法检测基质金属蛋白酶2(matrix metalloproteinases,MMP-2)、MMP-9、Toll样受体-4(TLR4)、IκB-α、p-IκB-α、NFκB和p-NFκB蛋白的表达。结果Tan IIA对LPS诱导的RAW264.7细胞培养液中炎症因子TNF-α、IL-6、IL^-1β和MCP-1的分泌有明显的抑制作用;明显下调MMP-2、MMP-9、TLR4、p-IκB-α和p-NFκB的蛋白的表达,抑制IκB-α磷酸化和NFκB的入核和活化。结论Tan IIA可通过抑制MMP-2和MMP-9的表达以及TLR4/κB-α/NF-κB信号通路,调控TNF-α、IL-6、IL^-1β等炎症因子的释放而发挥抗炎活性。     

Addition of tanshinone IIA to UW solution decreases skeletal muscle ischemia-reperfusion injury

AIM: To investigate whether tanshinone IIA could improve the effect of UW solution for skeletal muscle preservation and to determine the dose range of tanshinone IIA providing optimal protection during ischemia and reperfusion. METHODS: Ischemic rat limbs were perfused with UW solution or UW plus tanshinone IIA (UW+T, 0.05, 0.1, or 0.2 mg/mL) for 0.5 h before reperfusion; controls (I/R) received no perfusion. Serum creatine phosphokinase (CPK), aspartate aminotransferase (AST), and lactate dehydrogenase (LDH) were measured pre-ischemia and after reperfusion (2-h, 4-h, and 6-h). Muscle water content, superoxide dismutase (SOD), malondialdehyde (MDA), adenosine triphosphatase (ATPase) were assessed pre-reperfusion and after 6-h reperfusion. Intercellular adhesion molecule-1 (ICAM-1) and apoptosis were detected after 6-h reperfusion. Reperfusion blood flow was monitored during reperfusion period. RESULTS: UW and UW+T prevented luxury perfusion during reperfusion and inhibited ICAM-1 expression and apoptosis after 6-h reperfusion. Serum CPK, AST, and LDH levels in UW rats were significantly less than those in controls after 2-h reperfusion (no difference, 4-h or 6-h reperfusion). After 4-h ischemia, there were significant differences in water content, MDA, SOD, and ATPase between UW and controls, but no difference after 6-h reperfusion. All tests with UW+T rats were significantly different from control results at corresponding durations. Higher tanshinone doses improved results. CONCLUSION: UW plus tanshinone IIA increased protection against I/R injury, suggesting that tanshinone IIA has clinical value.     

Activation of CYP3A-mediated testosterone 6β-hydroxylation by tanshinone IIA and midazolam 1-hydroxylation by cryptotanshinone in human liver microsomes

This study evaluated the in vitro activation of CYP3A-mediated midazolam 1-hydroxylation and testosterone 6β-hydroxylation by tanshinone I, tanshinone IIA, and cryptotanshinone. The abilities of tanshinones to activate CYP3A-mediated midazolam 1-hydroxylation and testosterone 6β-hydroxylation in human liver microsomes (HLMs) were tested. Substrate- and effector-dependent activation of CYP3A by tanshinones were both observed. Cryptotanshinone was shown to activate CYP3A-mediated midazolam 1-hydroxylation in a concentration-dependent manner. In contrast, tanshinone IIA and tanshinone I did not activate this hydroxylation reaction. In addition, tanshinone IIA activated CYP3A-mediated testosterone 6β-hydroxylation, whereas cryptotanshinone and tanshinone I did not. The results from our study enhance the understanding of CYP3A activation by tanshinone IIA and cryptotanshinone in HLMs. Additionally, these data allow for an accurate prediction of the magnitude and likelihood of Danshen-drug interactions.     

PF2401-SF, standardized fraction of Salvia miltiorrhiza and its constituents, tanshinone I, tanshinone IIA, and cryptotanshinone, protect primary cultured rat hepatocytes from bile acid-induced apoptosis by inhibiting JNK phosphorylation.

Bile acid-induced hepatocyte apoptosis plays an important role in cholestatic liver disease, and the role of apoptosis may be of therapeutic interest in preventing liver disease. The dried root of Salvia miltiorrhiza Bunge (Labiatae) has been used traditionally to treat liver diseases. We investigated the antiapoptotic effects of a standardized fraction of S. miltiorrhiza (PF2401-SF) and its components, tanshinone I, tanshinone IIA, and cryptotanshinone, in primary cultured rat hepatocytes. PF2401-SF was enriched with tanshinone I (11.5%), tanshinone IIA (41.0%), and cryptotanshinone (19.1%). Glycochenodeoxycholic acid (GCDC)-induced apoptosis, as shown by DNA fragmentation, poly(ADP-ribose) polymerase cleavage, and activation of caspases-8, -9, and -3. PF2401-SF and its components, tanshinone I, tanshinone IIA, and cryptotanshinone showed antiapoptotic activity. Treatment with PF2401-SF or with its components significantly inhibited the generation of intracellular reactive oxygen species. Hydrophobic bile acids activate c-Jun-NH(2)-terminal kinase (JNK), p38 mitogen-activated protein kinases (MAPK), and extracellular signal-regulated kinase 1/2, and PF2401-SF inhibited the phosphorylation of JNK and p38. All three components of PF2401-SF inhibited JNK phosphorylation. Addition of inhibitors of MAPK showed that inhibition of JNK decreased apoptosis. These data indicate that PF2401-SF and its components protect hepatocytes from GCDC-induced apoptosis in vitro by inhibiting JNK.     

Induction of apoptosis by tanshinone I via cytochrome c release in activated hepatic stellate cells

Hepatic stellate cells play central roles in hepatic fibrosis. The therapeutic goal in hepatic fibrosis is to halt or reverse fibrosis. Apoptosis is suggested to eliminate activated hepatic stellate cells in fibrosis. Salvia miltiorrhiza is a traditional medicine used to improve blood circulation and treat chronic hepatitis and hepatic fibrosis. We investigated the effect of tanshinone I, an ingredient of Salvia miltiorrhiza, on the apoptotic death of rat hepatic stellate cells transformed by simian virus 40 (T-HSC/Cl-6), which retains the features of activated stellate cells. Treatment of T-HSC/Cl-6 cells with tanshinone I resulted in the induction of typical DNA fragmentation and DNA ladder formation in a concentration- and time-dependent manner. The induction of apoptosis was confirmed by flow cytometric analysis. Treatment of T-HSC/Cl-6 cells with tanshinone I caused activation of caspase-3 and subsequent proteolytic cleavage of poly(ADP-ribose) polymerase. Tanshinone I induced mitochondrial membrane dipolarization and the release of cytochrome c from mitochondria into the cytosol. In conclusion, our results demonstrate that tanshinone I induces apoptosis of T-HSC/Cl-6 cells and that tanshinone I-induced apoptosis involves caspase activation through cytochrome c release and loss of mitochondrial membrane potential.     

Simultaneous determination of danshensu,rosmarinic acid,cryptotanshinone, tanshinone IIA,tanshinone I and dihydrotanshinone I by liquid chromatographic–mass spectrometry and the application to pharmacokinetics in rats

A sensitive and specific liquid chromatography–tandem mass spectrometry method (LC–MS) was developed and validated for the separation and simultaneous determination of danshensu, rosmarinic acid and tanshinone compounds including cryptotanshinone, tanshinone I, dihydrotanshinone I and tanshinone IIA in rat plasma. Chromatographic separation of the analytes was successfully achieved on a C₁₈ column using a mobile phase composed of acetonitrile–water containing 0.5% glacial acetic acid. This method demonstrated good linearity and did not have endogenous material interfering with the active compounds and I.S. peaks. The limit of quantification of danshensu, rosmarinic acid, cryptotanshinone, dihydrotanshinone I, tanshinone I and tanshinone IIA were 5, 0.75, 0.1, 0.1, 1 and 0.5 ng/mL. The average extraction recoveries of these analytes from rat plasma were all over 60%. The precisions determined from five days were all within 10%. This method has been successfully applied in the simultaneous quantification and the pharmacokinetic studies of these six compounds in animals which were orally administered with danshen preparations.     

Tanshinone I enhances learning and memory,and ameliorates memory impairment in mice via the extracellular signal-regulated kinase signalling pathway

Background and purpose:

The intracellular signalling kinase, extracellular signal-regulated kinase 1/2 (ERK1/2) is required for new memory formation, suggesting that control of ERK signalling might be a target for the treatment of cognitive dysfunction. Previously, we reported that tanshinone congeners have ameliorating effects on drug-induced memory impairment in mice. Here, we have investigated possible modes of action of tanshinone I on learning and memory, associated with ERK phosphorylation.

Experimental approach:

Using immunohistochemical, Western blot techniques, and behavioural testing, we studied the effect of tanshinone I on memory impairment induced by diazepam or dizocilpine (MK-801) in mice.

Key results:

Tanshinone I (2 or 4 mg·kg⁻¹, p.o.) increased latency times versus vehicle-treated control group in the passive avoidance task. Western blot analysis and immunohistochemical data showed that tanshinone I (4 mg·kg⁻¹) increased levels of phosphorylated cAMP response element binding protein (pCREB) and phosphorylated ERK (pERK) in the hippocampus. These increases in pCREB and pERK were blocked by U0126 (inhibitor of ERK1/2), which also prevented the increase in passive avoidance task latency time after tanshinone I. In models of learning and memory impairment induced by diazepam and MK-801, tanshinone I (4 mg·kg⁻¹) reversed learning and memory impairments detected by the passive avoidance test. Western blot analysis showed that tanshinone I reversed the diazepam- and MK-801-induced inhibitions of ERK and CREB activation in hippocampal tissues. These effects were also blocked by U0126.

Conclusions and implications:

Tanshinone I ameliorates the learning and memory impairments induced by diazepam and MK-801 through activation of ERK signalling.     

Preventive effects of a purified extract isolated from Salvia miltiorrhiza enriched with tanshinone I,tanshinone IIA and cryptotanshinone on hepatocyte injury in vitro and in vivo

Salvia miltiorrhiza is traditionally used to treat liver disease in Asia. In this study, we tested the ability of a purified extract of S. miltiorrhiza (PF2401-SF) and its constituents, tanshinone I, tanshinone IIA, and cryptotanshinone, to protect against acute and subacute liver damage induced by carbon tetrachloride by measuring serum transaminase levels, the reduced form of glutathione (GSH), antioxidant enzyme activities, and lipid peroxidation levels in the liver. We also evaluated their ability to protect primary cultured rat hepatocytes from tertiary-butylhydroperoxide (tBH) or d-galactosamine (GalN). PF2401-SF was protective at 50–200 mg/kg per day in acute liver injury and 25–100 mg/kg per day in subacute liver injury. Tanshinone I, tanshinone IIA, and cryptotanshinon (40 μM), inhibited lactate dehydrogenase leakage, GSH depletion, lipid peroxidation and free radical generation in vitro. PF2401-SF and its major constituents, tanshinone I, tanshinone IIA and cryptotanshinone, can protect against liver toxicity in vivo and in vitro due to its antioxidant effects.