丹参酮ⅡA磺酸钠合成工艺研究

为改善丹参酮的水溶性,在其分子中引入水溶性官能团磺酸基.以不同的磺化剂与之发生磺化反应,结果表明,冰醋酸一浓硫酸的混合液是合成丹参酮水溶性衍生物的合适磺化剂.通过薄层色谱法、紫外吸收和红外吸收光谱法对其结构进行鉴定,证实了产物是丹参酮ⅡA磺酸钠.对以冰醋酸-浓硫酸为磺化剂合成丹参酮ⅡA磺酸钠进行了正交试验研究,得出其最佳工艺条件为:冰醋酸-浓硫酸(1:1,v/v),反应温度15℃,反应时间1 h,产品得率38.7%.     

藏药翼首草薄层鉴别方法研究

本文对藏药翼首草薄层鉴别方法进行系统考察。试验以熊果酸为对照品,采用单因素试验方法,对影响翼首草薄层色谱的因素进行了考察,筛选最佳薄层鉴别方法。结果采用硅胶G板,薄层板置展开缸中饱和10min,以10%硫酸乙醇溶液为显色剂,105℃加热至斑点清晰,置日光和紫外光灯(365 nm)下检视,所得的薄层色谱分离效果较好。研究结果表明本法分离效果好、重复性好,可用于藏药翼首草的质量控制。     

丹参有效成份的研究 Ⅲ.丹参酮Ⅱ—A 的资源寻找

前文报道丹参酮Ⅱ-A 磺酸钠是治疗冠心病的新类型药物。因此,从丹参同属植物特别是从民间的丹参类同品中,寻找丹参酮Ⅱ-A 高含量的种类就有一定的实际意义。现将各地采集的20种鼠尾草属(Salvia)植物的根,用95%酒精提取后,进行薄层定性检查,选择含有丹参酮Ⅱ-A等二萜醌化合物的10种植物样品,利用前报方法测定丹参酮Ⅱ-A、次甲丹参醌、丹参酮Ⅰ的含量,结果见表1。     

藏药婆婆纳药材薄层鉴别方法学研究

本文建立了藏药婆婆纳药材的薄层鉴别方法。采用单因素试验方法,对影响婆婆纳对照药材的薄层色谱因素进行系统考察,筛选出最佳的薄层色谱条件。该薄层鉴别条件吸附剂:硅胶G薄层板;展开剂:环己烷-丙酮(6∶4);显色剂:10%硫酸乙醇溶液;显色条件:110℃烘箱中加热至斑点显色清晰,日光下检视,所得的薄层色谱,斑点清晰,分离度好。该方法分离度高,重现性好,简便,可用于婆婆纳的质量控制。     

纯蛇粉胶囊中乌梢蛇的薄层层析鉴别

目的用薄层层析和紫外分析方法对纯蛇粉胶囊中所含的乌梢蛇成分进行分析鉴别。方法以中药乌梢蛇作为对照品,以硅胶Ｇ铺板,在丙酮、乙醇、氨水和正丁醇、乙醇、氨水２个系统中分别展开,后置于紫外２５４ｎｍ下检视,同法作空白试验。结果在同一薄层色谱中,和对照品相同的位置上,显清晰分离很好的４个斑点,其中２个斑点呈强烈的荧光。空白阴性对照品无此斑点。结论该方法具有定性检查的鉴别意义,为制定纯蛇粉胶囊质量标准提供了实验方法和依据。     

羟基丹参酮ⅡA的合成及体外抗肿瘤作用

对丹参酮ⅡA进行结构修饰,合成了羟基丹参酮ⅡA。采用MTT法考察了羟基丹参酮ⅡA对人宫颈癌细胞株Hela细胞,人肝癌细胞株HepG-2细胞、人胃癌细胞株SGC-7901细胞的增殖抑制作用。结果表明:羟基丹参酮ⅡA对三种肿瘤细胞增殖都有很好的抑制作用,抑制作用呈剂量依赖性。羟基丹参酮ⅡA对SGC-7901细胞抑制作用最强,其IC_(50)为4.18μM;对HeLa细胞的抑制作用次之,其IC_(50)为6.08μM;对HepG-2细胞抑制作用较弱,其IC_(50)为10.20μM。而丹参酮ⅡA对SGC-7901细胞、HeLa细胞和HepG-2细胞的IC_(50)分别是17.15μM、27.28μM和46.34μM。羟基丹参酮ⅡA抑制肿瘤细胞增殖作用明显强于丹参酮ⅡA(P0.05)。     

丹参酮Ⅱ-A在RAW264.7细胞系中的抗炎症作用机制

目的:丹参酮Ⅱ-A是丹参的一种重要成分。研究丹参酮Ⅱ-A的抗炎症机制。方法:以小鼠腹腔巨噬细胞系RAW264.7作为药物刺激靶细胞,使用不同浓度的分析纯丹参酮Ⅱ-A对RAW264.7细胞系进行刺激,在细胞被刺激24、48h后,使用MTT比色法和半定量RT-PCR法对刺激后的细胞进行检测。结果:通过MTT比色法检测,发现丹参酮Ⅱ-A抑制炎症细胞的增殖,初步计算出丹参酮Ⅱ-A的半抑制浓度(IC50)为43.2μmol/L;在半定量RT-PCR实验中发现,在发生炎症后,丹参酮Ⅱ-A通过抑制磷脂酶A2来减轻炎症。结论:在炎症发生后,丹参酮Ⅱ-A通过抑制磷脂酶A2来达到其抗炎症的作用。     

丹参酮ⅡA对大鼠移植性肝癌的抑制作用

目的:观察丹参酮Ⅱ A对大鼠移植性肝癌的抑制作用.方法:将SD大鼠随机分为假手术组、模型组、丹参酮ⅡA高、低剂量组.各组动物于移植术后12d处死.分离肿瘤测量其体积,realtime RT-PCR法和western blot法检测肿瘤组织中HMGB1、VEGFmRNA和蛋白的表达.结果:丹参酮ⅡA能明显降低肿瘤体积,降低肿瘤组织中HMGB1、VEGFmRNA和蛋白的表达.结论:丹参酮ⅡA能抑制大鼠移植性肝癌的生长,其机制可能与抑制HMGB1、VEGF表达有关.     

不同制剂中相同中草药薄层色谱鉴别探讨

目的探讨不同制剂中相同中药材统一鉴别的可行性。方法采用薄层色谱法,取不同制剂适量,按统一方法制备供试液、层析条件、显色与检视斑点。结果多种含大黄、人参的制剂,薄层色谱斑点清晰明显。结论本实验方法可行,在不同制剂中相同药材鉴别有较统一的方法为好。     

丹参酮ⅡA结构修饰及其对HeLa细胞的抑制作用研究

本研究运用Mannich反应,得到了丹参酮ⅡA的16位被酯化氨基酸取代的5个衍生物,结构均经EI-MS、1H及13C NMR确证。经MTT法测定丹参酮ⅡA及其衍生物对HeLa细胞增殖的抑制作用,并计算出IC50。结果表明,5个衍生物的生物活性较丹参酮ⅡA均有所增强;即丹参酮ⅡA在16位进行结构修饰可以增强其对肿瘤细胞抑制活性。     

The anti-atherosclerotic effect of tanshinone IIA is associated with the inhibition of TNF-α-induced VCAM-1, ICAM-1 and CX3CL1 expression

Tanshinone IIA is one of the major diterpenes in Salvia miltiorrhiza. The inhibitory effect of Tanshinone IIA on atherosclerosis has been reported, but the underlying mechanism is not fully understood. The present study aimed to study the anti-atherosclerosis effect of Tanshinone IIA on the adhesion of monocytes to vascular endothelial cells and related mechanism. Results showed that Tanshinone IIA, at the concentrations without cytotoxic effect, dose-dependently inhibited the adhesion of THP-1 monocytes to the TNF-α-stimulated human vascular endothelial cells. The expressions of cell adhesion molecules including VCAM-1, ICAM-1 and E-selectin were induced by TNF-α in HUVECs at both the mRNA and protein levels. The mRNA and protein expressions of VCAM-1 and ICAM-1, but not E-selectin, were both significantly suppressed by Tanshinone IIA in a dose dependent manner. In addition, the TNF-α-induced mRNA expression of fractalkine/CX3CL1 and the level of soluble fractalkine were both reduced by Tanshinone IIA. We also found that Tanshinone IIA significantly inhibited TNF-α-induced nuclear translocation of NF-κB which was resulted from the inhibitory effect of Tanshinone IIA on the TNF-α-activated phosphorylation of IKKα, IKKβ, IκB and NF-κB. As one of the major components of Salvia miltiorrhiza, Tanshinone IIA alone exerted more potent effect on inhibiting the adhesion of monocytes to vascular endothelial cells when compared with Salvia miltiorrhiza. All together, these results demonstrate a novel underlying mechanism for the anti-inflammatory effect of Tanshinone IIA by modulating TNF-α-induced expression of VCAM-1, ICAM-1 and fractalkine through inhibition of TNF-α-induced activation of IKK/NF-κB signaling pathway in human vascular endothelial cells.     

4种鼠尾草属植物根、茎、叶中脂溶性成分的分布差异性

为了说明4种鼠尾草属植物脂溶性成分在根、茎、叶中的分布情况,本文采用了组织化学定位和HPLC图谱分析相结合的方法,对4种鼠尾草属植物不同部位的脂溶性成分进行了分析.结果表明:4种鼠尾草属植物的根、茎、叶中脂溶性成分的分布有差异,根中脂溶性成分分布于周皮,茎、叶中脂溶性成分分布于表皮.丹参和白花丹参根中的脂溶性成分主要是丹参酮ⅡA、丹参酮Ⅰ和二氢丹参酮Ⅰ,鼠尾草和药用鼠尾草根中仅含丹参酮ⅡA,不含丹参酮Ⅰ和二氢丹参酮Ⅰ.茎、叶中的脂溶性成分不是丹参酮ⅡA、丹参酮Ⅰ和二氢丹参酮Ⅰ.脂溶性成分的种类和含量随品种和引种地的不同而产生差异.这一结果明确了脂溶性成分在4种鼠尾草属植物不同部位的确切分布,为准确用药提供了依据.     

Tanshinone IIA protects against cardiac hypertrophy via inhibiting calcineurin/NFATc3 pathway

Pathological cardiac hypertrophy induced by adrenergic overactivation can subsequently develop to heart failure which remains as a leading cause of mortality worldwide. Tanshinone IIA is a lipid-soluble pharmacologically active compound extracted from the rhizome of the Chinese herb Salvia miltiorrhiza, a well-known traditional Chinese medicine used for the treatment of cardiovascular disorders. However, little is know about the effect of Tanshinone IIA on cardiac hypertrophy. The present study was aimed to investigate whether Tanshinone IIA prevents cardiac hypertrophy induced by isoproterenol (ISO) and to clarify its possible mechanisms. Cardiomyocytes hypertrophy was induced by ISO 10 μM for 48 h with or without Tanshinone IIA 10, 30, 100 μM pretreatment, and evaluated by determining the cell size and the expression of ANP, BNP, β-MHC, Calcineurin, and NFATc3 by real-time PCR and western blot. We found that Tanshinone IIA pretreatment attenuated the enlargement of cell surface area induced by ISO in cultured cardiomyocytes. The mRNA level of ANP, BNP and β-MHC was obviously elevated in ISO-treated cardiac cells, which was effectively inhibited by Tanshinone IIA. Moreover, we found that Tanshinone IIA pretreatment could prevent the augment of intracellular calcium transient in ISO-treated cardiomyocytes. The further study revealed that Calcineurin, NFATc3, ANP, BNP and β-MHC proteins were upregulated by ISO in ventricular myocytes, and Tanshinone IIA pretreatment significantly attenuate the increased expression of Calcineurin, NFATc3, ANP, BNP and β-MHC proteins. In summary, Tanshinone IIA attenuated cardiomyocyte hypertrophy induced by ISO through inhibiting Calcineurin/NFATc3 pathway, which provides new insights into the pharmacological role and therapeutic mechanism of Tanshinone IIA in heart diseases.     

The relative importance of ultraviolet and longer wavelengths in the response properties of fly visual systems

A generalized analysis of the generator potential responses of R1-6 cells of Calliphora provides remarkable information on the visual properties for the Diptera. This shows that, although these cells have two peak response sensivities for monochromatic stimuli at 350 and 480 nm under single color stimulus conditions, and when the background illumination is either zero or in the region of 450–560 nm, the sensitivity to ultraviolet light is practically eliminated for background illumination in either the ultraviolet or the region around 600 nm or when any simultaneous dynamic stimulus in the region of 480–550 nm is also applied. These results seem somewhat perplexing to an understanding of the behavioral vision properties. It also is not consistant with the concept that the ultraviolet response is initiated by a sensitizing pigment within these cells that transfers energy to the rhodopsin-metarhodopsin process. However, it strengthens other evidence that the limited condition of ultraviolet responses comes from interaction from R7,8 cells but does not play an important behavioral role in the visual system fed from cells R1-6. As discussed in this paper, any high level pattern recognition controlling behavioral response to ultraviolet stimuli comes from the R7,8 cell system.     

丹参酮对心肌肌质网脂质过氧化过程中脂类自由基的清除作用

应用自旋捕集方法和化学发光方法研究天然抗氧化剂丹参酮（Ｔａｎｓｈｉｎｏｎｅ）对心肌肌质网膜脂质过氧化过程中产生的脂类自由基的清除作用。发现在一定的浓度范围内,丹参酮对脂质过氧化有较好的保护作用,在丹参酮浓度大于１ｍｇ／ｍｇ蛋白时,对脂类自由基清除率可达６０％。丹参酮对肌质网膜脂质过氧化的保护机理可能是通过清除脂类自由基而阻断脂质过氧化的链式反应,而不是清除氧自由基而防止脂质过氧化的启动。     

RAD6+ gene of Saccharomyces cerevisiae codes for two mutationally separable deoxyribonucleic acid repair functions.

The response of two mutant alleles of the RAD6+ gene of Saccharomyces cerevisiae to the ochre translational suppressor SUQ5 was determined. Both the ultraviolet sensitivity phenotype and the deficiency in ultraviolet-induced mutagenesis phenotype of the rad6-1 allele were suppressed in a [psi+] background. For the rad6-3 allele, only the ultraviolet-sensitivity phenotype was suppressible in a [psi+] background. An SUQ5 rad6-3 [psi+] strain that was examined showed the normal rad6-3 deficiency in ultraviolet-induced mutagenesis. We propose that the RAD6+ gene is divided into two cistrons, RAD6A and RAD6B. RAD6A codes for an activity responsible for the error-prone repair of ultraviolet-induced lesions in deoxyribonucleic acid but is not involved in a cell's resistance to the lethal effects of ultraviolet light. RAD6B codes for an activity essential for error-free repair of potentially lethal mutagenic damage.     

丹参酮IIA对大鼠移植性肝癌的抑制作用

目的：观察丹参酮IIA对大鼠移植性肝癌的抑制作用。方法：将sD大鼠随机分为假手术组、模型组、丹参酮[IA高、低剂量组。各组动物于移植术后12d处死。分离肿瘤测量其体积,reahimeRT．PCR法和western blot法检测肿瘤组织中HMGB1、VEGFmRNA和蛋白的表达。结果：丹参酮IIA能明显降低肿瘤体积,降低肿瘤组织中HMGBl、VEGFmRNA和蛋白的表达。结论：丹参酮IIA能抑制大鼠移植性肝癌的生长,其机制可能与抑制HMGB1、VEGF表达有关。     

Tanshinone IIA protects lens epithelial cells from H2O 2-induced injury by upregulation of lncRNA ANRIL

Tanshinone IIA is a lipophilic diterpene extracted from the Salvia miltiorrhiza bunge, possessing antiapoptotic and antioxidant activities. The purpose of this study was to explore the effects of Tanshinone IIA on age-related nuclear cataract. Human lens epithelial cell line SRA01/04 was subjected to H ₂O ₂ to mimic a cell model of cataract. Cell Counting Kit-8 assay, flow cytometer, and reactive oxygen species (ROS) detection were performed to evaluate the effect of Tanshinone IIA pretreatment on SRA01/04 cells injured by H ₂O ₂. Besides, the real-time quantitative polymerase chain reaction was used to assess the expression of long noncoding RNA (lncRNA) antisense noncoding RNA in the INK4 locus (ANRIL). Western blot analysis was performed to detect the expression of core proteins involved in cell survival and nuclear factor-κB (NF-κB) pathway. H ₂O ₂ significantly decreased SRA01/04 cells viability, whereas increased apoptosis and ROS generation. This phenomenon was coupled with the upregulated p53, p21, Bax, cleaved caspase-3, and the downregulated cyclinD1, CDK4, and Bcl-2. Tanshinone IIA pretreatment protected SRA01/04 cells against H ₂O ₂-induced injury. In the meantime, the expression of lncRNA ANRIL was upregulated by Tanshinone IIA. And, the protective effects of Tanshinone IIA on H ₂O ₂-stimulated SRA01/04 cells were abolished when lncRNA ANRIL was silenced. Moreover, the elevated expression of lncRNA ANRIL induced by Tanshinone IIA was abolished by BAY 11-7082 (an inhibitor of NF-κB). To conclude, Tanshinone IIA protects SRA01/04 cells from apoptosis triggered by H ₂O ₂. Tanshinone IIA confers its protective effects possibly via modulation of NF-κB signaling and thereby elevating the expression of lncRNA ANRIL.     

Tanshinone IIA from Salvia miltiorrhiza BUNGE inhibits human aortic smooth muscle cell migration and MMP-9 activity through AKT signaling pathway

Smooth muscle cell (SMC) migration plays an important role in normal angiogenesis and is relevant to disease-related vascular remodeling in conditions such as brain arteriovenous malformations, pulmonary hypertension, arteriosclerosis, and restenosis after angioplasty. In this present study, we showed that tanshinone IIA, the major lipid-soluble pharmacological constituent of Salvia miltiorrhiza BUNGE, inhibits human aortic smooth muscle cell (HASMC) migration and MMP-9 activity. Tanshinone IIA significantly inhibited IkappaBalpha phosphorylation and p65 nuclear translocation through inhibition of AKT phosphorylation. Tanshinone IIA inhibited TNF-alpha-induced ERK and c-jun phosphorylation, but not other MAPKs such as JNK and p38. Tanshinone IIA also inhibited NF-kappaB and AP-1 DNA-binding. Moreover, tanshinone IIA inhibited the migration of TNF-alpha-induced HASMCs. Our results provide evidence that tanshinone IIA has multiple effects in the inhibition of HASMC migration and may offer a therapeutic approach to block HASMC migration.