Danshen (Salvia miltiorrhiza) extract inhibits proliferation of breast cancer cells via modulation of Akt activity and p27 level

Danshen is widely used in traditional Chinese medicine, often in combination with other herbs. To check the effect of Danshen on the proliferation of breast cancer cells, Danshen extract was used to treat MCF‐7 and MCF‐7 HER2 cells, the latter of which overexpresses HER2. HER2 is a receptor tyrosine kinase, and is involved in signal transduction pathways leading to tumor cell proliferation. MTT and cell proliferation assays revealed that Danshen strongly inhibited the proliferation of both MCF‐7 vec cells and MCF‐7 HER2 cells. Flow cytometry analyses indicated that Danshen induced cell cycle delay in the G1 phase. HER2 expression was shown to confer resistance to Danshen‐induced inhibition of proliferation and cell cycle delay, suggesting that HER2 is responsible for the resistance to Danshen. Danshen treatment induced the down‐regulation of Akt phosphorylation and an increase in p27 in MCF‐7 vec and MCF‐7 HER2 cells. Nevertheless, MCF‐7 HER2 cells were more resistant to the Danshen‐induced inhibition of Akt phosphorylation and p27 up‐regulation. Copyright © 2009 John Wiley & Sons, Ltd.     

盐酸小檗碱与环孢素A合用对小鼠肝药物代谢酶的影响

目的：阐明盐酸小檗碱与环孢素A合用对药物代谢酶的影响，方法：采用分光光度法测定盐酸小檗碱，环孢素A及二者合用时小鼠肝微粒体CYP450，ERD，ADM和GST的含量或活性。结果：ig给药3和6d，盐酸小檗碱（200mg/kg)对CYP450，ERD没有明显的抑制作用，但对ADM和GSP有抑制作用，环孢素A（45mg/kg)对ERD，AMD和GST均有抑制作用；盐酸小檗碱与环孢素A合用对CYP450，ERD，ADM和GST均有明显抑制作用。结论：盐酸小檗碱与环孢素A的组合是药物代谢酶CYP450，ERD（CYP3A），ADM（CYP1A1，2B1，2C11）和GST的强抑制剂，其总体抑制水平至少与已知的抑制剂酮康唑和肖苯地平相当甚至更强。     

扶正化瘀方对正常及肝纤维化大鼠体内CYP450酶的影响

使用Cocktail探针药物法,研究扶正化瘀方(FZHY)对正常和肝纤维化大鼠细胞色素P450(CYP450)5种同工酶的影响。正常和二甲基亚硝胺(dimethylnitrosamine,DMN)腹腔注射诱导的肝纤维化模型大鼠,以FZHY浸膏粉连续灌胃2周后,单剂量尾静脉注射由CYP450 5种同工酶的特异性探针底物非那西丁(CYP1A2)、甲苯磺丁脲(CYP2C9)、奥美拉唑(CYP2C19)、氢溴酸右美沙芬(CYP2D6)和咪达唑仑(CYP3A4)配置的Cocktail探针溶液后,通过建立的LC-MS/MS法同时检测血浆中5种探针药物的浓度,采用PK solution 2TM对数据进行处理,并计算主要药动学参数。正常大鼠灌胃FZHY后,非那西丁、甲苯磺丁脲、奥美拉唑和氢溴酸右美沙芬的AUC0-t均出现不同程度的增加,CL也均出现降低趋势,提示FZHY能明显抑制正常大鼠CYP1A2,CYP2C9,CYP2C19,CYP2D6的活性,但对CYP3A4的活性影响不明显;肝纤维化大鼠灌胃FZHY后,甲苯磺丁脲的AUC0-t显著增加,Vd显著降低,其他4种探针药物的药动学参数未见显著改变,提示在肝纤维化大鼠体内FZHY仅对CYP2C9有抑制作用,对CYP1A2,CYP2C19,CYP2D6,CYP3A4活性影响不明显。CYP450酶在肝纤维化条件下活性的改变可能是FZHY对正常和肝纤维化大鼠CYP450酶影响不同的原因。     

Exploring the possible metabolism mediated interaction of Glycyrrhiza glabra extract with CYP3A4 and CYP2D6

The rhizome of Glycyrrhiza glabra L. (licorice) is used very widely in Indian and Chinese traditional medicine, and it is a popular flavor ingredient of drinks, sweets and candies. Its medicinal uses include treating bronchitis, dry cough, respiratory infections, liver disorders and diabetes. Glycyrrhizin is normally considered to be its biologically active marker, so a rapid RP‐HPLC method was developed for the quantitative estimation of glycyrrhizin in the extract. The effect of the standardized extract and its marker on drug metabolizing enzymes was evaluated through CYP3A4 and CYP2D6 inhibition assays to evaluate the safety through its drug interaction potential. The inhibition of CYP3A4 and CYP2D6 isozymes was analysed by the fluorescent product formation method. In the CYP450‐CO assay, the interaction potential of the standardized extract and pooled microsomes (percentage inhibition 23.23 ± 1.84%), was found to be less than the standard inhibitor. In the fluorimetric assay, G. glabra extracts showed higher IC₅₀ values than their positive inhibitors, ketoconazole and quinidine for CYP3A4 and CYP2D6, respectively. Furthermore, the interaction potential of the plant extract was greater than the pure compound. The results demonstrate that G. glabra and its principle bioactive compound, glycyrrhizin, when co‐administered with conventional medicines showed only a weak interaction potential with drug metabolizing enzymes. Copyright © 2011 John Wiley & Sons, Ltd.     

红花注射液对大鼠细胞色素P450 2D6亚型的抑制作用

目的研究红花注射液对大鼠细胞色素P450 2D6亚型(CYP2D6)的影响。方法利用探针药物右美沙芬(DM),高效液相色谱法测定各实验组(对照组,红花注射液0.9、1.8、3.6 mL/kg组)大鼠体内尿液中与体外肝微粒体温孵系统中DM的代谢率,考察红花注射液对大鼠CYP2 D6活性的影响。结果体内实验和体外实验中,红花注射液1.8和3.6 mL/kg组DM的代谢率均明显低于对照组(P<0.05、0.01);抑制实验中,体外肝微粒体温孵系统中红花注射液组和西咪替丁组DM的代谢率明显低于空白组(P<0.05);含红花生药量为30 mg/mL时红花注射液组DM的代谢率与西咪替丁(0.6 mg/mL)时DM的代谢率相近,抑制能力相当;红花注射液的IC50为10.64 mg/mL。结论红花注射液对大鼠CYP2D6有显著的抑制作用。     

丹参提取液对球形红细菌生长的影响

目的 考察丹参提取液对球形红细菌(光合细菌)生长的影响.方法 通过添加不同质量浓度的常规培养基和丹参提取液,培养球形红细菌,考察球形红细菌的活菌数、生长曲线、脱氢酶活性及光合色素吸收光谱,研究丹参提取液对球形红细菌生长的影响.结果 培养基中添加丹参提取液后可使球形红细菌活菌数增加到之前的1.6倍,并缩短球形红细菌生长的延迟期,提前进入指数期及稳定期,可使球形红细菌脱氧酶活性提高到之前的6.6倍.低质量浓度丹参提取液基本不影响球形红细菌中光合色素的形成,高质量浓度时吸收均变低,且影响类胡萝卜素的形成,其红外光谱855 nm处红移至863 nm.结论 培养基中添加丹参提取液可影响球形红细菌生长,缩短球形红细菌生长周期,提高球形红细菌的活力,高质量浓度时影响光合色素的形成.     

植物外源激素对丹参生长和丹参酮类物质积累的影响

目的研究植物外源激素赤霉素(GA3)和吲哚乙酸(IAA)对丹参生长和丹参酮类物质积累的影响。方法采用温室石英砂盆栽试验与室内分析相结合的方法。结果单独施用GA3有利于丹参地上部分生物量的增加,但对地下部分生物量表现一定抑制作用;施用低浓度和高浓度GA3均有利于3种丹参酮(隐丹参酮、丹参酮Ⅰ、丹参酮ⅡA)的累积,施用中等浓度GA3则不利于3种丹参酮的累积。单独施用IAA对丹参地上部分和地下部分生物量的影响呈现先升后降趋势,但统计学差异不显著,且对株高、根长的影响亦不显著;总体来看,施用低质量浓度IAA(0.5mg/L)有利于丹参根中3种丹参酮的累积,但随着IAA处理浓度增加,仅隐丹参酮量有所增加,而丹参酮Ⅰ和丹参酮ⅡA量反而下降。两种激素配施时,低浓度GA3与低浓度IAA组合处理对丹参的生长表现出显著促进作用,并能显著提高丹参根中3种丹参酮的量。结论施用适量的植物外源激素GA3和IAA能够促进丹参的生长和丹参根中3种丹参酮类物质的积累。     

丹参酚酸A对大鼠肝微粒体细胞色素P450酶系的影响

目的:研究丹参酚酸A对大鼠肝微粒体细胞色素P450和细胞色素b_5含量以及CYP1A2和CYP2E1活性的影响.方法:将大鼠分成溶剂对照组和丹参酚酸A给药组,每组10只,雌雄各半,丹参酚酸A给药组尾静脉注射给予丹参酚酸A 20 mg·kg~(-1)·d~(-1),连续给药5 d;溶剂对照组给予相同剂量的溶剂,紫外分光光度法测定大鼠肝微粒体细胞色素P450和细胞色素b_5含量;探针底物法评价CYP1A2和CYP2E1的活性.结果:丹参酚酸A尾静脉注射连续给药5 d后,大鼠细胞色素P450和细胞色素b_5含量与对照组比较均无显著性差异;CYP1A2和CYP2E1的活性与对照组比较也无显著性差异.结论:丹参酚酸A对CYP1A2和CYP2E1没有诱导或抑制作用,与经过CYP1A2和CYP2E1代谢的药物发生相互作用的可能性较小.     

Antidiabetic effect of the total polyphenolic acids fraction from Salvia miltiorrhiza Bunge in diabetic rats

An investigation was made to evaluate the therapeutic potential of the total polyphenolic acids fraction (PAF) from Salvia miltiorrhiza Bunge in the type 2 diabetes mellitus rats model with an oral dose of 187 mg/kg for 28 days. The results showed that PAF induced a significant decrease in fasting blood glucose (FBG), fasting blood insulin (FINS), total cholesterol (TC), triglyceride (TG) and blood urea nitrogen (BUN), and an obvious increase in insulin sensitivity index (ISI) in diabetic rats induced by a high fat diet and a low dose of streptozocin (STZ). These results suggested that PAF has antidiabetic potential in vivo. Copyright © 2011 John Wiley & Sons, Ltd.     

丹参的体外抑菌作用研究

目的：探讨丹参的体外抑菌作用。方法：用K-B纸片扩散法。100％丹参浸出液滤纸片对大肠杆菌、金黄色葡萄球菌、白色葡萄球菌、变形杆菌、乙型链球菌抑菌作用进行了研究。结果：丹参对以上细菌均有抑菌作用。结论：丹参在体外有明显的抑菌作用。     

A randomized, double-blind, placebo-controlled study to evaluate the efficacy and tolerability of Fufang Danshen (Salvia miltiorrhiza) as add-on antihypertensive therapy in Taiwanese patients with uncontrolled hypertension

Hypertension generally requires the use of a combination therapy to achieve the satisfactory control of blood pressure. A traditional Chinese herb, Danshen (Salvia miltiorrhiza), has been shown to have cardioprotective effects in animals and humans. The study investigated the add‐on effect of Fufang Danshen extract capsule in Taiwanese hypertensive patients with uncontrolled blood pressure. This was a double‐blind, placebo‐controlled, randomized, single‐center study clinical trial. Fifty‐five patients with uncontrolled mild to moderate hypertension were enrolled under current conventional antihypertensive treatment, randomized equally to receive a Fufang Danshen capsule (formula mixture) 1000 mg twice‐daily or a placebo capsule for 12 weeks. Primary endpoints were the control rate and the response rate. By ITT analysis at week 12, the control rates were 25.5% in the Fufang Danshen group and 7.3% in the control group (p = 0.016). The response rates were 45.6% in the Fufang Danshen group and 38.2% in the placebo group (p = 0.946). A significant reduction of systolic blood pressure at week 12 was noted in the Fufang Danshen group compared with the placebo group (13.8 vs 4.2 mmHg, p = 0.005). A decrease of pulse rate was also noted in the Fufang Danshen group (? 3.2 vs +2.7/min, p = 0.027). Adverse events were not statistically different between the two groups. It was concluded that Fufang Danshen (Salvia miltiorrhiza) extract reduced systolic blood pressure and pulse rate, and was well tolerated in patients with hypertension. Copyright © 2011 John Wiley & Sons, Ltd.     

丹参抑制泼尼松大鼠骨质丢失

目的:探讨长期超生理剂量应用泼尼松对大鼠所致的骨质疏松症特点,同时观察中药丹参对其防治作用。方法:24只♂SD大鼠随机分成正常对照组,泼尼松组和丹参组,每组8只。正常对照组给予生理盐水处理,其他组先予泼尼松按2.7 mg/kg ig,随后泼尼松组灌喂生理盐水,丹参治疗组按5.0 g/kg灌喂丹参水提液。每日1次,连续12周。实验结束后用骨组织形态计量学等方法测量股骨的动态参数和静态参数,同时测量尺骨的羟脯胺酸和钙磷含量以及测量股骨的长度和宽度。结果:泼尼松大鼠骨量显著丢失及骨生长受到抑制(P<0.01),而丹参治疗组可明显对抗泼尼松对骨的生长抑制作用。结论:4月龄SD大鼠服用泼尼松12周后可造成骨质疏松症,丹参对其有较好的对抗作用。     

单次不同剂量PEG400对大鼠体内细胞色素P450 3A活性的影响

 目的研究单次不同剂量聚乙二醇(PEG)400对大鼠体内细胞色素P4503A活性的影响。方法以咪哒唑仑为探针,HPLC测定咪哒唑仑及其代谢物1′-羟基咪哒唑仑在大鼠体内的血药浓度并计算其药动学参数,以不同剂量PEG400处理组与阴性对照组的AUC_0-4h比值为指标,研究它们对大鼠体内细胞色素P4503A药酶活性的影响。结果与生理盐水组相比,PEG400(15mg·kg^-1)、PEG400(60mg·kg^-1)分别显著增加咪哒唑仑AUC_0-4h 1.98,1.75倍(P<0.05),显著降低1′-羟基咪哒唑仑与咪哒唑仑AUC_0-4h的比值,分别从1.09降至0.26和0.28(P<0.05)。结论两种剂量PEG400对CYP3A均有明显的抑制作用。     

探针药物咪哒唑仑有限采样法预测肝损伤大鼠 CYP 3A 代谢活性的研究

 目的 以细胞色素 P450 （ CYP ） 3A 探针药物咪哒唑仑（ MDZ ）的系统清除率（ CL _s ）为指标,评价有限采样法（ LSS ）预测肝脏损伤状态下 CYP 3A 代谢活性的可行性。 方法 采用系列浓度的 四氯化碳溶液 预处理大鼠, 24 h 后,静脉注射 MDZ ,在若干时间点采血检测血浆 MDZ 浓度。留取血清并检测 丙氨酸氨基转移酶（ ALT ）和天冬氨酸氨基转移酶（ AST ）活性 。经逐步回归分析和 Jack-knife 方法 验证,建立最终的 LSS 模型。对经相同处理的另一随机群体进行验证分析,评价该 LSS 模型方程的准确性和重现性。 结果 系列浓度四氯化碳溶液造成肝脏不同程度损伤。由单点（ 45 min ）或两点（ 5 , 4 5 min ）血浆药物浓度建立的 LSS 预测模型所得到的 CL _s 估计值（ CL _est ）与实际计算值（ CL _obs ）之间具有良好的相关性,误差小。两点 LSS 模型对样本预测的相关性较单点 LSS 更优（ r =0.96 ）,而单点 LSS 模型则更显简便。 结论 本实验表明,以 MDZ 清除率为指标,采用 45 min 或 5. 4 5 min 的 有限采样方案评价肝脏损伤状态下 CYP 3A 的代谢活性是一种准确而简便的方法,为今后推广到临床评价肝脏代谢功能从而制定和调整给药方案提供了理论依据和实验室证据。     

决明子水提液对大鼠肝脏CYP450酶的影响

该文研究决明子水提液对大鼠肝脏CYP450酶酶活性、mRNA及蛋白表达水平的影响。取SD大鼠,口服灌胃受试药物后,处死,用冰冷生理盐水灌流肝脏,提取大鼠肝脏微粒体、肝脏总RNA和总蛋白,采用Cocktail体外孵育法结合液质联用(LCMS)技术对大鼠肝脏中CYP1A2,2B1,2C11,2D2,2E1,3A1各亚酶的酶活性进行测定,并应用荧光定量PCR技术和Western blot对上述亚型的mRNA和蛋白表达水平进行检测。结果表明,酶活性方面,决明子水提液组与空白组比较可明显诱导CYP1A2,2B1,2C11,2D2,2E1,3A1的酶活性,其中决明子低剂量组对CYP2D2有明显的抑制作用,中、高剂量组对CYP2D2有明显的诱导作用,且都呈剂量依赖性增加,但对CYP3A1的诱导不呈剂量依赖性;mRNA表达方面,决明子水提液对CYP1A2,2C11,2D2,2E1mRNA的表达具有显著诱导作用,其中对CYP1A2,2D2,2E1诱导作用呈剂量依赖性,与酶活性水平具有一致性,对CYP2B1,3A1没有明显作用;蛋白表达方面,决明子水提液对CYP2C11,2E1的表达也具有诱导作用,但没有统计学差异。决明子水提液对CYP450同工酶不同程度诱导或抑制作用,特别是对于CYP2C11,2E1亚型酶,酶活性与mRNA,蛋白表达水平具有一致性,提示当与这些酶底物合并用药时,尤其是与CYP2C11,2E1代谢有关的药物合用时,应充分考虑到潜在的有益和不利的药物相互作用。     

合成冰片对大鼠肝CYP450酶含量及CYP3A1 mRNA表达的影响

[目的]研究冰片对大鼠肝脏细胞色素氧化酶（cytochromeP450,CYP450）含量及其亚型CYP3AImRNA表达的影响。[方法] Wistar大鼠合成冰片（设0．3g／kg和0.03g／kg剂量组,1次／d,连续7d）灌胃,以溶媒羧甲基纤维素纳（CMC-Na）和苯巴比妥钠为对照组,钙沉积法提取肝微粒体,紫外分光光度法测定CYP450含量,实时定量RT-PCR法检测肝脏CYP3AImRNA的表达。[结果]0．3g／kg合成冰片口服可诱导大鼠肝脏CYP450酶含量增高（P〈0．05）,CYP3A1 mRNA表达上调（P〈0．05）。[结论]高剂量合成冰片口服可能影响肝脏药物代谢。     

大鼠肝药酶活性改变对紫杉醇药动学的影响

 目的 通过考察CYP3A的变化对大鼠体内紫杉醇代谢的影响,评价紫杉醇与其它可能合用的药物之间的相互作用。方法 本研究将135只♀Wistar大鼠随机分入空白组、诱导组或抑制组。各组CYP3A的水平由于使用诱导剂或抑制剂而有所不同。大鼠尾静脉注射紫杉醇10 mg·kg^-1后,采集动态血浆标本,以HPLC测定血药浓度,采用PCNONLIN程序进行房室模型数据拟合。结果 大鼠CYP3A被诱导和抑制时,紫杉醇代谢发生了明显的改变。空白对照组,诱导剂组和抑制剂组的c_max分别为68.91,56.51和108.53 μg·ml^-1,AUC为82.48,53.96和189.47 μg·h·ml^-1,而CL则为0.0242,0.037和0.0105 L·h^-1。结伦 大鼠CYP3A亚族在紫杉醇生物转化过程中起着重要作用。对酶作用的认识有利于预测并控制药物不良反应和可能的药物相互作用。     

Hypolipidemic activity and mechanism of purified herbal extract of Salvia miltiorrhiza in hyperlipidemic rats

AIM OF THE STUDY: The study aimed at evaluating the hypolipidemic effects of Purified Salvia miltiorrhiza extract (PSME) and investigating the potential molecular mechanisms by which PSME modulated lipid profiles in hyperlipidemic rats. MATERIALS AND METHODS: Sprague-Dawley male rats on a high-fat/high-cholesterol diet were treated orally with PSME, GW3965 (a selective liver X receptor agonist) or vehicle alone. Gene expression analysis and transactivation assays were used to clarify the molecular mechanisms of action of PSME. RESULTS: The concentrations of plasma total cholesterol, low-density lipoprotein cholesterol (LDL-cholesterol) and triglycerides in rats treated with PSME at 150mgkgday(-1) were significantly decreased (P<0.01), accompanied with significantly decreased concentrations of liver total cholesterol and triglycerides (P<0.01). In both drug-treated rats, the concentration of high-density lipoprotein cholesterol (HDL-cholesterol) was significantly elevated (P<0.01). Intriguingly, short heterodimer partner (SHP) mRNA level was significantly higher in PSME-treated rats (P<0.01), accompanied with the significantly decreased mRNA level of sterol regulatory element binding protein 1c (SREBP1c) (P<0.01), which contributed to the decreases of liver and plasma triglycerides through a farnesoid X receptor-SHP-SREBP1c pathway. ATP-binding Cassette Transporter B11 (ABCB11) and murine Mdr2 P-glycoprotein (also known as ABCB4) were significantly induced by PSME, which were responsible for biliary cholesterol solubility by proper biliary secretion of bile salts and phospholipids. The transactivation assays were used to identify PSME as a farnesoid X receptor/liver X receptor alpha coagonist. CONCLUSION: These results indicated that PSME as a farnesoid X receptor/liver X receptor alpha coagonist largely improved the lipid profiles in the hyperlipidemic rats.     

Antiproliferative activity and apoptosis induction by trijuganone C isolated from the root of Salvia miltiorrhiza Bunge (Danshen)

In this study, we found that the hexane fraction of Danshen, the dried root of Salvia miltiorrhiza (Lamiaceae), exerted antiproliferative effects on human leukemia cells. Phytochemical investigation of the hexane fraction achieved the isolation of the tanshinone diterpenes: dihydrotanshinone I ( 1 ), trijuganone C ( 2 ), trijuganone B ( 3 ), cryptotanshinone ( 4 ), tanshinone IIA ( 5 ), and tanshinone I ( 6 ). Compound 2 showed significant antiproliferative activities against human leukemia cells HL‐60, Jurkat, and U937. The antiproliferative activities of 2 against human cancer and normal cells indicated that 2 exhibited potent antiproliferative activities with IC₅₀ values less than 10 μM against HL‐60 and Jurkat cells as well as on the colon cancer cells DLD‐1, COLO 205, and Caco‐2. Compound 2 induced chromatin condensation, DNA fragmentation, activation of caspase‐3, ‐8, and ‐9, and the cleavage of poly (ADP‐ribose) polymerase (PARP) in HL‐60 cells. Moreover, 2 activated Bid and Bax, leading to the loss of mitochondrial membrane potential, and 2 induced the cytochrome c release from mitochondria into cytosol. In contrast, Bcl‐2 and Bcl‐xL were unaffected by 2 . These results suggest that 2 exerts antiproliferative effects via apoptosis induction mediated by mitochondrial dysfunction and caspase activation. Compound 2 may serve as a candidate of potential chemotherapeutic agent for human leukemia.     

壮骨关节丸对大鼠肝微粒体P450的影响

[目的]考察壮骨关节丸对大鼠肝微粒体细胞色素P450的影响。[方法]壮骨关节丸及其两个新工艺按含生药2.1 g/kg连续给大鼠灌胃7 d,末次药后24 h断头处死大鼠并取肝脏,用钙沉降法制备肝微粒体。在体外用含氨苯砜、非那西丁、奥美拉唑、氯唑沙宗的cocktail探针代谢来考察肝微粒体CYP3A、CYP1A2、CYP2C19、CYP2E1的活性。[结果]cocktail探针在体外肝微粒体系统中代谢1 h后,包括壮骨关节丸及其两个新工艺的给药组剩余氯唑沙宗浓度显著高于对照组,而奥美拉唑、非那西丁、氨苯砜浓度与对照组之间差异无统计学意义。[结论]壮骨关节丸可以抑制大鼠肝脏CYP2E1活性,但对CYP1A2、CYP3A及CYP2C19无显著影响。