迷迭香二萜酚类成分鼠尾草酸的研究进展

迷迭香(Rosmarinus officinalis L.)系唇形科迷迭香属多年草本植物,研究表明迷迭香中主要的二萜酚类成分鼠尾草酸(carnosic acid)除了具有抗氧化作用,还具有抗癌、神经保护、抗炎、减肥、抑菌等多种药理活性,表现出较高的药用价值,并且可以广泛应用于药品、保健品、化妆品、食品添加剂及食用油等行业,为了促进鼠尾草酸的新药研究和开发利用,本文对其近年来的研究进展进行综述。     

RP-HPLC法测定迷迭香中鼠尾草酸和迷迭香酸的含量

目的建立高效液相色谱法同时测定迷迭香药材中鼠尾草酸和迷迭香酸的含量。方法运用PhenomenexC18（250 mm×4.60 mm,5μm）为分析柱;柱温30℃;流动相为乙腈（A）-0.1%磷酸（B）梯度洗脱：0～10min,25%A,10～15 min：25%～75%A,15～25 min：75%A;流速1.0 mL min^-1;检测波长284 nm。结果迷迭香酸和鼠尾草酸与其他组分的分离效果好,两者分别在0.046 55～1.862 0μg和0.372 7～14.908μg呈良好的线性关系（r值为0.999 94和0.999 97）,平均回收率分别为99.0%（RSD=1.1%,n=9）和98.1%（RSD=1.5%,n=9）。结论该测定方法简便、准确、重现性好,方便了迷迭香药材中鼠尾草酸和迷迭香酸的含量测定。     

迷迭香中二萜酚化合物的抗氧化作用

目的：研究迷迭香中二萜酚化合物鼠尾草酚、迷迭香酚和表迷伫香酚对人低密度脂蛋白氧化的抑制作用及其对自由基和超氧阴离子的清除作用。方法：以细胞膜和LDL为靶,用TBARS方法和ESR方法研究了抗氧化活性。用荧光方法研究对铜诱导的LDL中apo B蛋白氧化的抑制。结果：鼠尾草酚、迷迭香酚和表迷迭香酚对人血LDL中的脂质过氧化和apo B蛋白的氧化均有抑制作用,IC50值在7－10μmol/L。抗氧化机理与其对脂自由基清除活性有关。结论：鼠尾草酚、迷迭香酚和表迷迭香酚能抑制LDL氧化。     

Determination of carnosic acid in Rosmarinus officinalis L. using square wave voltammetry and electrochemical behavior

A new, fast, sensitive and simple voltammetric method is established for the direct determination of carnosic acid (CA). And the electroreduction of carnosic acid (CA) was studied using electrochemical methods. The number of electrons transferred in electrode mechanisms were calculated for reversible and adsorption-controlled electrochemical reduction of CA at 17 mV versus Ag/AgCl at pH 7.0 in Britton–Robinson buffer (BR) on a hanging mercury drop electrode. Square-wave voltammetry was developed and validated for direct determination of CA. Square-wave parameters were optimized as accumulation potential = 0.0 mV, accumulation time = 5 s, frequency = 50 Hz, pulse amplitude = 50 mV, and staircase step potential = 5 mV. The developed method displays three linear responses from 2 to 9 μM, 10 to 30 and 40 to 90 μM for carnosic acid with a correlation coefficient of 0.996, 0.999 and 0.999. The detection limits were found to be 1.5 μM, 4.0 μM and 40.1 μM, respectively. The interference effect of most common organic and inorganic species was investigated. Proposed method was successfully applied for determination of CA in natural extract of rosemary and the average content was determined as 11.9 ± 1.0 (μg CA/1 g rosemary). The results were in agreement with that obtained by HPLC-UV comparison method. The developed method can be widely used in routine quality control of herbal materials as well as other in foods, medicinal, pharmaceutical and environmental analysis.     

Antiplatelet aggregation activity of diterpene alkaloids from Spiraea japonica

Six diterpene alkaloids with an atisine-type C(20)-skeleton isolated from the Chinese herbal medicines Spiraea japonica var. acuta and S. japonica var. ovalifolia, as well as eight derivatives of spiramine C and spiradine F were evaluated for the ability to inhibit aggregation of rabbit platelets induced by arachidonic acid, ADP, and platelet-activating factor (PAF) in vitro. The results showed that 12 of the 14 atisine-type diterpene alkaloids significantly inhibited PAF-induced platelet aggregation in a concentration-dependent manner, but had no effect on ADP- or arachidonic acid-induced aggregation, exhibiting a selective inhibition. It is the first report that C(20)-diterpene alkaloids inhibit PAF-induced platelet aggregation. However, spiramine C1 concentration-dependently inhibited platelet aggregation induced by PAF, ADP and arachidonic acid with IC(50) values of 30.5+/-2.7, 56.8+/-8.4 and 29.9+/-9.9 microM, respectively, suggesting a non-selective antiplatelet aggregation action. The inhibitory effect of spiramine C1 on arachidonic acid was as potent as that of aspirin. Primary studies of the structure-activity relationships for inhibition of PAF-induced aggregation showed that the oxygen substitution at the C-15 position and the presence of an oxazolidine ring in spiramine alkaloids were essential to their antiplatelet aggregation effects. These results suggest that the atisine-type alkaloids isolated from S. japonica are a class of novel antiplatelet aggregation agents.     

迷迭香的三萜类化学成分研究

从昆明产的迷迭香（Rosmarinus officinalis,L.)的茎叶中分离得到五个三萜类化合物（I－Ⅴ）,经光谱等方法鉴定其结构为：α－香树素二十六烷酸脂（α－amyrin hexacosoatewa,I),α-白檀酮（α－amyrenone,Ⅱ）,桦木酸（betulinic acid,Ⅲ）,齐墩果酸（oleanolic acid,Ⅳ）,熊果酸（ursolic acid,V);其中I为一个新三萜。     

HPLC-DAD phenolic profile,cytotoxic and anti-kinetoplastidae activity of Melissa officinalis

Context Melissa officinalis subsp. inodora Bornm. (Lamiaceae) has been used since ancient times in folk medicine against various diseases, but it has not been investigated against protozoa.

Objective To evaluate the activities of M. officinalis against Leishmania braziliensis, Leishmania infantum and Trypanosoma cruzi as well as its cytotoxicity in fibroblast cell line.

Materials and methods The fresh leaves were chopped into 1?cm² pieces, washed and macerated with 99.9% of ethanol for 72?h at room temperature. Antiparasitic activity of M. officinalis was accessed by direct counting of cells after serial dilution, while the cytotoxicity of M. officinalis was evaluated in fibroblast cell line (NCTC929) by measuring the reduction of resazurin. The test duration was 24?h. High-performance liquid chromatography (HPLC) was used to characterise the extract.

Results The extract at concentrations of 250 and 125?μg/mL inhibited 80.39 and 54.27% of promastigote (LC_50? value?=_?105.78?μg/mL) form of L. infantum, 80.59 and 68.61% of L. brasiliensis (LC₅₀ value _?=_?110.69?μg/mL) and against epimastigote (LC₅₀ value _?=_?245.23?μg/mL) forms of T. cruzi with an inhibition of 54.45 and 22.26%, respectively, was observed. The maximum toxicity was noted at 500?μg/mL with 95.41% (LC_50? value?=_?141.01?μg/mL). The HPLC analysis identified caffeic acid and rutin as the major compounds.

Discussion The inhibition of the parasites is considered clinically relevant (<?500?μg/mL). Rutin and caffeic acids may be responsible for the antiprotozoal effect of the extract.

Conclusion The ethanol extract of M. officinalis can be considered a potential alternative source of natural products with antileishmania and antitrypanosoma activities.     

Anti-inflammatory activity of abietic acid, a diterpene isolated from Pimenta racemosa var. grissea.

The anti-inflammatory activity of abietic acid, a diterpene isolated from Pimenta racemosa var. grissea (Myrtaceae), was evaluated in-vivo and in-vitro. This compound significantly inhibited rat paw oedema induced by carrageenan in a time- and dose-dependent manner, and mouse ear oedema induced by 12-O-tetradecanoylphorbol acetate, after oral or topical administration. The inhibition of myeloperoxidase enzyme showed that its topical activity was influenced by neutrophil infiltration into the inflamed tissues (ears). In addition, the effect of abietic acid on some macrophage functions was analysed in-vitro. Non-toxic concentrations of abietic acid inhibited prostaglandin E2 (PGE2) production in lipopolysaccharide-treated macrophages, whereas nitrite, tumour necrosis factor alpha and interleukin-1beta production were only weakly affected by this diterpene. PGE2 production from A23187-stimulated macrophages was only inhibited at high doses (100 microM) and it failed to modify leukotriene C4 production. These results indicate that abietic acid exerts in-vivo anti-inflammatory activity after oral or topical administration and has partial ability to prevent the production of some inflammatory mediators.     

A new phenolic glycoside and a new trans-clerodane diterpene from Conyza blinii

A new phenolic glycoside, 4-propionyl-2,6-dimethoxyphenyl beta-D-glucopyranoside (1) and a new trans-clerodane diterpene named 19-deacetylconyzalactone (2), were isolated from the aerial parts of Conyza blinii.     

Anti-inflammatory activity of sugiol, a diterpene isolated from Calocedrus formosana bark

Sugiol is a diterpene which was isolated and purified from alcohol extracts of the bark of Calocedrus formosana Florin (Cupressaceae). Although sugiol has low inhibitory activity against the DPPH radical, it could effectively reduce intracellular reactive oxygen species (ROS) production in lipopolysaccharide (LPS)-stimulated macrophages. The present study investigated the potential anti-inflammatory activity of sugiol, and the relationship between signal transduction and inflammatory cytokines in vitro. A dose of 30 microM of sugiol was effectively inhibitory for proIL-1beta, IL-1beta and TNF-alpha production, suggesting that sugiol is bioactive against inflammation. Moreover, sugiol reveals a capacity for suppressing the activation of mitogen-activated protein kinases (MAPKs), including extracellular signal-regulated kinase (ERK), c-Jun NH2-terminal kinase (JNK), and p38 mitogen-activated protein kinase (p38) activated by LPS-stimulation in J774A.1 murine macrophages. A low dosage of 10 microM of sugiol completely inhibited ERK1/2 phosphorylation, while 30 microM effectively inhibited JNK1/2 and p38 phosphorylation in LPS-stimulated macrophages. In addition, sugiol significantly inhibited LPS-induced ROS production. Our studies suggest that sugiol's efficacy in inhibiting the inflammatory cytokines of IL-1beta and TNF-alpha could be attributed to a reduction of the ROS that leads to a decrease in the phosphorylation of MAPKs.     

Efficacy of Rosmarinus officinalis leaves extract against cyclophosphamide-induced hepatotoxicity

Context Cyclophosphamide (CTX) is used to treat different cancer types, although it causes severe hepatotoxicity due to its oxidative stress effect. Rosmarinus officinalis, L. (Lamiaceae) has a therapeutic potential against hepatotoxicity due to its antioxidant activity.

Objective The objective of this study is to investigate the phytochemical analysis of the methanol extract of Rosmarinus officianalis leaves (MEROL) and its efficacy against CTX-induced hepatotoxicity.

Materials and methods The phytochemical analyses were assessed spectrophotometericaly. To assess the MEROL efficacy, 72 Swiss albino mice were divided into six groups. Group 1 was control, groups 2 and 3 included mice which were injected intraperitoneally (i.p.) with 100 or 200?mg/kg of MEROL at days 1, 4, 7, 10, 13 and 16; group 4 was injected (i.p.) with CTX (200?mg/kg) at day 17, groups 5 and 6 were injected (i.p.) with MEROL as groups 3 and 4 followed by 200?mg/kg CTX at day 17, respectively. At day 22, six mice from each group were sacrificed and the others were sacrificed at day 37.

Results MEROL has a high content of total phenolics, saponins, total antioxidant capacity and DPPH radical scavenging activity. The median lethal dose (LD₅₀) value of MEROL was 4.125?g/kg b.w. The inhibitory concentration 50 (IC₅₀) value for DPPH radical scavenging was 55?μg/mL. Pretreatment with 100?mg/kg MEROL for 16 d ameliorated CTX-induced hepatotoxicity represented in lowering the levels of the aspartate aminotransferase (AST) and lipid profile and minimizing the histological damage.

Conclusions Pretreatment with 100?mg/kg b.w. MEROL mitigated CTX-induced hepatotoxicity due to its antioxidant activity.     

In Vitro Production of Essential Oil from Proliferating Shoots of Rosmarinus officinalis1

The 40-day-old IN VITRO proliferating shoots of ROSMARINUS OFFICINALIS L. var. GENUINA forma ERECTUS produced an appreciable quantity of essential oil, i.e., 1.8% fw, which was similar in its constituents to that obtained from 1-year-old plants, whether naturally grown or IN VITRO-raised potted plants. However, the quantity of various constituents identified so far was generally, but marginally, less in the former case than the latter two kinds of 1-year-old plants with the exception of bornyl acetate and 1,8-cineole, the concentrations of which were higher in the proliferated shoots than the plants. The essential oil content of 1-year-old naturally grown plants was 2.4% fw, while it was 2.38% fw in the IN VITRO-raised potted plants of the same age.     

Gastroprotective mechanisms of centipedic acid, a natural diterpene from Egletes viscosa LESS

This study was aimed to clarify the mechanisms of gastroprotection by centipedic acid (CPA), a natural diterpene from Egletes viscosa LESS. (Asteraceae) using ethanol-induced gastric mucosal damage in mice and gastric secretion in 4-h pylorus-ligated rats as model systems. In mice, intragastrically administered CPA (25, 50, 100 mg/kg) greatly reduced the mucosal lesions induced by 96% ethanol (0.2 ml, p.o.) by 18, 53, and 79%, respectively, whereas N-acetylcysteine (NAC, 300 mg/kg, i.p.), the reference compound produced a 50% inhibition. In 4-h pylorus-ligated rats, CPA (50 mg/kg) applied intraduodenally decreased both gastric secretory volume and total acidity. Similar to NAC, the plant diterpene effectively prevented the ethanol associated decrease in non-proteic sulfhydryls (NP-SH) and the elevated thiobarbituric acid-reactive substances (TBARS) in gastric tissue, suggesting that these compounds exert an antioxidant effect. Pretreatment of mice with indomethacin, the cyclooxygenase inhibitor but not with capsazepine, the transient receptor potential vanilloid-1 (TRPV1)-receptor antagonist greatly suppressed the gastroprotective effect of CPA. Furthermore, CPA gastroprotection was significantly attenuated in mice pretreated with L-NAME or glibenclamide the respective inhibitors of nitric oxide synthase and K(+)(ATP) channel activation. These data suggest that CPA affords gastroprotection by different and complementary mechanisms, which include a sparing effect on NP-SH reserve, and roles for endogenous prostaglandins, nitric oxide, and TRPV1-receptor and K(+)(ATP) channel activation.     

In vivo analgesic and anti-inflammatory activities of Rosmarinus officinalis aqueous extracts,rosmarinic acid and its acetyl ester derivative

Abstract

Context: Despite several pharmacological applications of Rosmarinus officinalis L. (Lamiaceae), studies on its analgesic and anti-inflammatory properties have been scarce.

Objective: The aim of this work was to use in vivo models to evaluate the analgesic and anti-inflammatory activities of the aqueous extracts obtained from leaves (AEL) and stems (AES) of Rosmarinus officinalis, as well as its isolated compound – rosmarinic acid (RA). We also prepared and assessed the acetyl ester derivative of RA.

Materials and methods: The analgesic activity was evaluated using abdominal constriction and formalin tests. For the evaluation of the anti-inflammatory effects, carrageenin-induced paw edema in rats were used. The extracts were used at doses of 100, 200 and 400?mg?kg^?1 compounds were tested at 10, 20 and 40?mg?kg^?1.

Results: Orally administered AEL, AES and RA were not significantly active at any of the doses tested during the abdominal constriction test; the acetyl ester derivative of RA displayed significant analgesic activity. In the carrageenin-induced paw edema assay, the acetyl derivative of RA at all the tested doses produced significant anti-inflammatory effects and reduced the number of paw licks in the second phase of the formalin test.

Discussion and conclusion: The results suggest that the analgesic effects of the acetyl derivative of RA operate via a peripheral-mediated mechanism. The acetyl ester derivative of RA is potentially applicable as a new lead compound for the management of pain and inflammation.     

鼠尾草酸的遗传毒性研究

目的 研究鼠尾草酸(carnosic acid,CA)的遗传毒性。方法 选用鼠伤寒沙门杆菌回复突变(Ames 试验)、体内哺乳动物红细胞微核、精子畸形以及单细胞凝胶电泳(彗星试验)四项致突变生物学试验进行筛选评价。结果 在6.25～50μg/mL 的剂量水平,CA 对鼠伤寒沙门菌株TA97、TA98、TA100、TA102 和TA1535 均无诱变性;此外,在受试剂量下小鼠骨髓嗜多染红细胞微核、小鼠精子畸形以及体内彗星试验的结果均为阴性(与溶剂对照比较,P> 0.05)。结论 在本实验条件下,CA 未见明显遗传毒性。     

Antiplatelet and antithrombotic effects of the diterpene spiramine Q from Spiraea japonica var. incisa

Spiramine Q, a diterpene, was isolated from a Chinese herbal plant Spiraea japonica var. incisa Yu. Born's and Wan HY's methods were used to investigate effects of spiramine Q on rabbit platelet aggregation and serotonin release, respectively. Its antithrombotic effect in mice was also evaluated by Myers' method. Spiramine Q selectively inhibited arachidonic acid-induced platelet aggregation in vitro or ex vivo, and decreased serotonin secretion from rabbit platelets. Spiramine Q (5 mg/kg) decreased the mouse mortality caused by injection of 80 mg/kg arachidonic acid in the tail vein. The results suggested that spiramine Q showed potent antiplatelet and antithrombotic activites.     

Characterization of the bactericidal activity of the natural diterpene kaurenoic acid

Kaurenoic acid is a diterpene with selective antibacterial activity against Gram-positive bacteria. The compound is bacteriolytic for Bacillus cereus. This activity was only partially affected by the composition and pH of the culture medium. Loss of the ability to retain the Gram stain and morphological alterations were produced in B. cereus cells exposed to kaurenoic acid. On the other hand, LPS mutants of Salmonella typhi were resistant to the compound, but spheroplasts of Escherichia coli became more sensitive to kaurenoic acid.