Citral,a component of lemongrass oil,activates PPARα and γ and suppresses COX-2 expression

Lemongrass is a widely used herb as a food flavoring, as a perfume, and for its analgesic and anti-inflammatory purposes; however, the molecular mechanisms of these effects have not been elucidated. Previously, we identified carvacrol from the essential oil of thyme as a suppressor of cyclooxygenase (COX)-2, a key enzyme for prostaglandin synthesis, and also an activator of peroxisome proliferator-activated receptor (PPAR), a molecular target for “lifestyle-related” diseases. In this study, we evaluated the essential oil of lemongrass using our established assays for COX-2 and PPARs. We found that COX-2 promoter activity was suppressed by lemongrass oil in cell-based transfection assays, and we identified citral as a major component in the suppression of COX-2 expression and as an activator of PPARα and γ. PPARγ-dependent suppression of COX-2 promoter activity was observed in response to citral treatment. In human macrophage-like U937 cells, citral suppressed both LPS-induced COX-2 mRNA and protein expression, dose-dependently. Moreover, citral induced the mRNA expression of the PPARα-responsive carnitine palmitoyltransferase 1 gene and the PPARγ-responsive fatty acid binding protein 4 gene, suggesting that citral activates PPARα and γ, and regulates COX-2 expression. These results are important for understanding the anti-inflammatory and anti-lifestyle-related disease properties of lemongrass.     

Inhibition of lipopolysaccharide-induced expression of inducible nitric oxide synthase by butein in RAW 264.7 cells

Butein has been reported to exert anti-inflammatory effect but the possible mechanism involved is still unclear. Here, we report the inhibitory effect of butein on nitric oxide (NO) production and inducible nitric oxide synthase (iNOS) gene expression. Butein also inhibited the induction of tumor necrosis factor-alpha and cyclooxygenase 2 by LPS. To further investigate the mechanism responsible for the inhibition of iNOS gene expression by butein, we examined the effect of butein on LPS-induced nuclear factor-kappaB (NF-kappaB) activation. The LPS-induced DNA binding activity of NF-kappaB was significantly inhibited by butein, and this effect was mediated through inhibition of the degradation of inhibitory factor-kappaB and phosphorylation of Erk1/2 MAP kinase. Furthermore, increased binding of the osteopontin alphavbeta3 integrin receptor by butein may explain its inhibitory effect on LPS-mediated NO production. Taken together, these results suggest that butein inhibits iNOS gene expression, providing possible mechanisms for its anti-inflammatory action.     

Anti-inflammatory activity of c-phycocyanin in lipopolysaccharide-stimulated RAW 264.7 macrophages

C-phycocyanin (C-PC), found in blue green algae, is often used as a dietary nutritional supplement. C-PC has been found to have an anti-inflammatory activity and exert beneficial effect in various diseases. However, little is known about its mechanism of action. Overproduction of nitric oxide (NO) derived from inducible nitric oxide synthase (iNOS) plays an important role in the pathogenesis of inflammation. The aim of this study was to determine whether C-PC inhibits production of nitrite, an index of NO, and iNOS expression in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages. Our results indicated that C-PC significantly inhibited the LPS-induced nitrite production and iNOS protein expression accompanied by an attenuation of tumor necrosis factor-alpha (TNF-alpha) formation but had no effect on interleukin-10 production in macrophages. Furthermore, C-PC also suppressed the activation of nuclear factor-kappaB (NF-kappaB) through preventing degradation of cytosolic IkappaB-alpha in LPS-stimulated RAW 264.7 macrophages. Thus, the inhibitory activity of C-PC on LPS-induced NO release and iNOS expression is probably associated with suppressing TNF-alpha formation and nuclear NF-kappaB activation, which may provide an additional explanation for its anti-inflammatory activity and therapeutic effect.     

椪柑精油及其主要抑菌组分对菌核青霉的抑制作用

采用水蒸气蒸馏法提取椪柑(Citrus reticulate Blanco)果皮精油,并用GC-MS对其成分进行分析,共鉴定出46种成分,主要包括柠檬烯(57.67%)、β-芳樟醇(5.36%)、2-蒈烯(4.47%)、β-蒎稀(4.31%)、γ-松油烯(4.08%)、α-蒎烯(3.27%)、1,2-二异丙烯基环丁烷(2.87%)、β-月桂烯(2.77%)、α-侧柏烯(2.40%)、β-水芹烯(2.24%)、癸醛(1.80%)和香茅醇(1.49%)等。采用污染食物技术(poisoned food technique)和液体培养法(liquid culture)测定了不同浓度椪柑精油以及主要抑菌组分对菌核青霉的抑制作用。结果表明:不同浓度(0.5~20μl/ml)的椪柑精油对菌核青霉生长均有一定的抑制,且抑菌效果与浓度呈正相关。无论是采用污染食物技术还是液体培养法,20μl/ml椪柑精油均能完全抑制菌核青霉生长;随着培养 时间延长到7d, 20μl/ml椪柑精油抑菌效果有所下降,但仍能显著抑制菌核青霉生长。采用污染食物技术考察了椪柑精油中7种常见抑菌成分对菌核青霉的影响。结果表明,0.04μl/ml柠檬醛和1.07μl/ml β-芳樟醇能显著抑制菌核青霉生长,而其他组分无明显作用。实验结果表明,椪柑精油的抑菌作用可能归功于其所含的柠檬醛和β-芳樟醇。     

Heparin-binding epidermal growth factor-like growth factor inhibits cytokine-induced NF-kappa B activation and nitric oxide production via activation of the phosphatidylinositol 3-kinase pathway

NO produced by inducible NO synthase (iNOS) has been implicated in various pathophysiological processes including inflammation. Therefore, inhibitors of NO synthesis or iNOS gene expression have been considered as potential anti-inflammatory agents. We have previously demonstrated that heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF) decreases proinflammatory cytokine IL-8 and NO production in cytokine-stimulated intestinal epithelial cells by interfering with the NF-kappaB signaling pathway. However, the upstream signaling mechanisms involved in these responses have not yet been defined. In this report, we show that in intestinal epithelial cells, HB-EGF triggered PI3K-dependent phosphorylation of Akt. Inhibition of PI3K reversed the ability of HB-EGF to block NF-kappaB activation, expression of iNOS, and NO production. Small interfering RNA of PI3K also reversed the inhibitory effect of HB-EGF on iNOS expression. Alternatively, transient expression of constitutively active PI3K decreased NO production by approximately 2-fold more than treatment with HB-EGF alone. This PI3K effect was HB-EGF dependent. Thus, activation of PI3K is essential but not sufficient for decreased NO synthesis. PI3K and HB-EGF act synergistically to decrease NO synthesis. Neither overexpression or inhibition of MEK, Ras, or Akt affected HB-EGF-mediated inhibition of NF-kappaB activation. These data demonstrate that HB-EGF decreases proinflammatory cytokine-stimulated NF-kappaB activation and NO production via activation of the PI3K signaling pathway. These results also suggest that inhibition of NF-kappaB and activation of the PI3K-dependent signaling cascade by HB-EGF may represent key signals responsible for the anti-inflammatory effects of HB-EGF.     

苦橙叶精油化学成分及抗炎作用研究

为探究苦橙叶精油的抗炎作用。实验采用气相-质谱联用法(GC-MS)分析精油成分,并建立脂多糖(LPS)诱导RAW 264.7细胞炎症模型,用Griess法检测一氧化氮(NO)含量评价其体外抗炎作用,随后进一步通过巴豆油致小鼠耳肿胀模型和鸡蛋清致小鼠足肿胀模型评价其体内抗炎作用。结果表明苦橙叶精油成分以酯类、醇类物质为主;25μg/mL浓度能显著抑制RAW 264.7细胞NO的释放;中浓度苦橙叶精油能明显减轻小鼠耳肿胀程度;低、中、高浓度苦橙叶精油均对小鼠足肿胀模型有炎症缓解作用,并于肿胀前期呈浓度依赖性。以上实验证明苦橙叶精油在体外和体内具有一定抗炎作用。     

Chemical Composition and Antimicrobial Activity of the Essential Oil from the Edible Aromatic Plant Aristolochia delavayi

The essential oil obtained by hydrodistillation from the aerial parts of Aristolochia delavayi Franch. (Aristolochiaceae), a unique edible aromatic plant consumed by the Nakhi (Naxi) people in Yunnan, China, was investigated using GC/MS analysis. In total, 95 components, representing more than 95% of the oil composition, were identified, and the main constituents found were (E)‐dec‐2‐enal (52.0%), (E)‐dodec‐2‐enal (6.8%), dodecanal (3.35%), heptanal (2.88%), and decanal (2.63%). The essential oil showed strong inhibitory activity (96% reduction) of the production of bacterial volatile sulfide compounds (VSC) by Klebsiella pneumoniae, an effect that was comparable with that of the reference compound citral (91% reduction). Moreover, the antimicrobial activity of the essential oil and the isolated major compound against eight bacterial and six fungal strains were evaluated. The essential oil showed significant antibacterial activity against Providencia stuartii and Escherichia coli, with minimal inhibitory concentrations (MIC) ranging from 3.9 to 62.5 μg/ml. The oil also showed strong inhibitory activity against the fungal strains Trichophyton ajelloi, Trichophyton terrestre, Candida glabrata, Candida guilliermondii, and Cryptococcus neoformans, with MIC values ranging from 3.9 to 31.25 μg/ml, while (E)‐dec‐2‐enal presented a lower antifungal activity than the essential oil.     

Astaxanthin inhibits nitric oxide production and inflammatory gene expression by suppressing I(kappa)B kinase-dependent NF-kappaB activation

Astaxanthin, a carotenoid without vitamin A activity, has shown anti-oxidant and anti-inflammatory activities; however, its molecular action and mechanism have not been elucidated. We examined in vitro and in vivo regulatory function of astaxanthin on production of nitric oxide (NO) and prostaglandin E2 (PGE2) as well as expression of inducible NO synthase (iNOS), cyclooxygenase-2, tumor necrosis factor-alpha (TNF-alpha), and interleukin-1beta (IL-1beta). Astaxanthin inhibited the expression or formation production of these proinflammatory mediators and cytokines in both lipopolysaccharide (LPS)-stimulated RAW264.7 cells and primary macrophages. Astaxanthin also suppressed the serum levels of NO, PGE2, TNF-alpha, and IL-1beta in LPS-administrated mice, and inhibited NF-kappaB activation as well as iNOS promoter activity in RAW264.7 cells stimulated with LPS. This compound directly inhibited the intracellular accumulation of reactive oxygen species in LPS-stimulated RAW264.7 cells as well as H2O2-induced NF-kappaB activation and iNOS expression. Moreover, astaxanthin blocked nuclear translocation of NF-kappaB p65 subunit and I(kappa)B(alpha) degradation, which correlated with its inhibitory effect on I(kappa)B kinase (IKK) activity. These results suggest that astaxanthin, probably due to its antioxidant activity, inhibits the production of inflammatory mediators by blocking NF-kappaB activation and as a consequent suppression of IKK activity and I(kappa)B-alpha degradation.     

山鸡椒挥发油成分分析及其抗真菌保鲜作用的研究

本文对湖南山鸡椒（Ｌｉｔｓｅａ ｃｕｂｅｂａ）挥发油（山苍子油）抑制植物病原真菌Ｆｕｓａｒｉｕｍ ｏｘｙｓｐｏｒｕｍ,Ｈｅｌｍｉｎｔｈｏｓｐｏｒｉｕｍ ｓｐ．Ｓｔｅｍｐｈｙｌｌｉｕｍ ｓｐ．的效能及其抑制真菌的有效成分进行了研究,实验结果表明,所用山鸡椒挥发油具有很强的抑制植物病原真菌生长的能力,并有明显的保鲜作用,其主要生物活性成份为柠檬醛。     

Chemical Composition and in vitro Biological Activities of the Essential Oil of Vepris macrophylla (Baker) I.Verd. Endemic to Madagascar

Vepris macrophylla is an evergreen tree occurring in sub‐humid forest of Madagascar and traditionally used in the Island to treat several complaints as well as to prepare aromatic teas and alcoholic drinks. In the present work, the essential oil distilled from the leaves was analyzed for the first time by gas chromatography (GC‐FID) and gas chromatography/mass spectrometry (GC/MS). The major compounds were citral (56.3%), i.e., mixture of neral (23.1%) and geranial (33.2%), citronellol (14.5%), and myrcene (8.3%). The essential oil exhibited antimicrobial activity against S. aureus, P. aeruginosa, and C. albicans as determined by vapor‐diffusion assay, supporting the traditional use of the plant for preparing steam bath for the treatment of infectious diseases. The essential oil was evaluated for cytotoxic activity on human tumor cell lines by MTT (=3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl‐2H‐tetrazolium bromide) assay, showing inhibitory effects comparable to those of cisplatin, notably on MDA‐MB 231 (human breast adenocarcinoma) and HCT116 (human colon carcinoma) cell lines. Finally, the essential oil was also subjected to screening for its antioxidant activity and the free radical scavenging capacity.     

Inhibitory effect of curcumin on nitric oxide production from lipopolysaccharide-activated primary microglia

Curcumin has been shown to exhibit anti-inflammatory, antimutagenic, and anticarcinogenic activities. However, the modulatory effect of curcumin on the functional activation of primary microglial cells, brain mononuclear phagocytes causing the neuronal damage, largely remains unknown. The current study examined whether curcumin influenced NO production in rat primary microglia and investigated its underlying signaling pathways. Curcumin decreased NO production in LPS-stimulated microglial cells in a dose-dependent manner, with an IC(50) value of 3.7 microM. It also suppressed both mRNA and protein levels of inducible nitric oxide synthase (iNOS), indicating that this drug may affect iNOS gene expression process. Indeed, curcumin altered biochemical patterns induced by LPS such as phosphorylation of all mitogen-activated protein kinases (MAPKs), and DNA binding activities of nuclear factor-kappaB (NF-kappaB) and activator protein (AP)-1, assessed by reporter gene assay. By analysis of inhibitory features of specific MAPK inhibitors, a series of signaling cascades including c-Jun N-terminal kinase (JNK), p38 and NF-kappaB was found to play a critical role in curcumin-mediated NO inhibition in microglial cells. The current results suggest that curcumin is a promising agent for the prevention and treatment of both NO and microglial cell-mediated neurodegenerative disorders.     

Direct scavenging of nitric oxide by traditional crude drugs.

Thirty-one traditional crude drugs and several pure compounds were examined for their possible regulatory effect on nitric oxide (NO) levels using sodium nitroprusside as a NO donor in vitro. Most of the crude drugs tested demonstrated direct scavenging of NO. Eight crude drugs, including Sanguisorbae Radix, Caryophylli Flos, Gambir, Coptidis Rhizoma, Granati Cortex, Gallae Rhois, Rhei Rhizoma and Cinnamomi Cortex exhibited significant activity (IC50 values < 1000 micrograms/ml), and with the exception of Coptidis Rhizoma, all were found to contain tannins as their major constituents. In addition, some crude drugs containing flavonoids or essential oils also appeared to act against NO. Ten major tannins contained in Sanguisorbae Radix and Rhei Rhizoma showed high scavenging activity (IC50 values < 326.3 micrograms/ml), and 6 of 8 alkaloids obtained from Coptidis Rhizoma also effectively scavenged the NO radical (IC50 values < 455.4 micrograms/ml). It was indicated that these compounds may be the active principles of the crude drugs responsible for NO scavenging. The present results suggest that traditional crude drugs might be potent and novel therapeutic agents for scavenging of NO and the regulation of pathological conditions caused by excessive NO and its oxidation product, peroxynitrite. These findings may also help to explain, at least in part, certain pharmacological activities of crude drugs, especially anti-infection and anti-inflammatory activities.     

Kojyl thioether derivatives having both tyrosinase inhibitory and anti-inflammatory properties

This study was conducted to examine the tyrosinase inhibitory and anti-inflammatory activities of kojic acid derivatives. A series of kojic acid derivatives containing thioether, sulfoxide, and sulfone linkages were synthesized. In the tyrosinase assay, kojyl thioether derivatives containing appropriate lipophilic alkyl chains (pentane, hexane, and cyclohexane) showed potent inhibitory activity. However, sulfoxides and sulfones exhibited decreased activity. Similar experimental results were obtained with inhibitory activities of NO production being induced by LPS. The presence of thioether linkage and appropriated lipophilic acid moiety was critical for the tyrosinase inhibitory and anti-inflammatory activities.     

锦鸡儿属分析生物地理学的研究

将锦鸡儿属（Caragana）植物分布区划分成13个。在分支系统学基础上,进行了分布区的成分分析。以种类和系为分布特征,进行分布区的聚类分析和最小生成树分析。分布区分支图、表征图和最小生成树从不同方面表达了分布区的关系。锦鸡儿属的分布区被分成东亚和古地中海两大部分。其中分别来自这两部分的前苏联远东-我国东北与阿尔泰-萨彦岭分布区有密切关系。基于分布区分支图,属的分布区分别由这两个分布区衍生,同时结合属的种系发生关系,推断本属可能起源于东西伯利亚,时间为第三纪中新世末-上新世。生态适应上,分类群是由东部温带中生性类型向西部旱化和寒化方向发展的。     

Insulin-like growth factor-II, phosphatidylinositol 3-kinase, nuclear factor-kappaB and inducible nitric-oxide synthase define a common myogenic signaling pathway.

Insulin-like growth factors (IGFs) are potent inducers of skeletal muscle differentiation and phosphatidylinositol (PI) 3-kinase activity is essential for this process. Here we show that IGF-II induces nuclear factor-kappaB (NF-kappaB) and nitric-oxide synthase (NOS) activities downstream from PI 3-kinase and that these events are critical for myogenesis. Differentiation of rat L6E9 myoblasts with IGF-II transiently induced NF-kappaB DNA binding activity, inducible nitric-oxide synthase (iNOS) expression, and nitric oxide (NO) production. IGF-II-induced iNOS expression and NO production were blocked by NF-kappaB inhibition. Both NF-kappaB and NOS activities were essential for IGF-II-induced terminal differentiation (myotube formation and expression of skeletal muscle proteins: myosin heavy chain, GLUT 4, and caveolin 3), which was totally blocked by NF-kappaB or NOS inhibitors in rat and human myoblasts. Moreover, the NOS substrate L-Arg induced myogenesis in the absence of IGFs in both rat and human myoblasts, and this effect was blocked by NOS inhibition. Regarding the mechanisms involved in IGF-II activation of NF-kappaB, PI 3-kinase inhibition prevented NF-kappaB activation, iNOS expression, and NO production. Moreover, IGF-II induced, through a PI 3-kinase-dependent pathway, a decrease in IkappaB-alpha protein content that correlated with a decrease in the amount of IkappaB-alpha associated with p65 NF-kappaB.