Anti-inflammatory and anti-allergic effect of rosmarinic acid (RA); inhibition of seasonal allergic rhinoconjunctivitis (SAR) and its mechanism

The present study was undertaken to determine whether oral supplementation with rosmarinic acid (RA) is an effective intervention for patients with SAR. In addition, the anti-inflammatory mechanism of RA also estimated in the ear edema models. CLINICAL TRIAL: Patients were treated daily with RA (200 mg or 50 mg) or placebo for 21 days. Patients recorded symptoms daily and profiles of infiltrating cells and concentration of cytokines were measured in nasal lavage fluid. Compared to placebo, supplementation with RA resulted in a significant decrease in responder rates for each symptom. RA also significantly decreased the numbers of neutrophils and eosinophils in nasal lavage fluid. ANIMAL STUDY: Topical application RA showed anti-inflammatory activity 5-hours after 12-tetradecanoylphorbol 13-acetate (TPA) treatment with marked inhibition of neutrophil infiltration. Up regulation of ICAM-1, VCAM-1 cyclooxygenase-2 (COX-2), KC and MIP-2 by TPA were markedly reduced by pre-treatment with extract of perilla (PE) or RA. Reactive oxygen radical production detected as thiobarbituric acid reactive substance (TBARS), lipid peroxide (LPO) and 8-hydroxy-2'deoxyguanosine (8OH-dG), by double treatment of TPA was reduced by pretreatment with PE or RA. RA is an effective intervention for SAR that is mediated by inhibition of PMNL infiltration. This effect of RA is due to two independent mechanisms: inhibition of the inflammatory response and scavenging of ROS.     

Rosmarinic acid,a major polyphenolic component of Perilla frutescens,reduces lipopolysaccharide (LPS)-induced liver injury in D-galactosamine (D-GalN)-sensitized mice

The protective activity of rosmarinic acid from Perilla frutescens on liver injury induced by LPS in D-GalN-sensitized mice was examined. We also investigated the effects of antitumor necrosis factor-alpha antibody (anti-TNF), superoxide dismutase (SOD), and aminoguanidine (AG) on this model in order to elucidate the mechanism of rosmarinic acid protection. Perilla extract (PE) and rosmarinic acid (RA) treatments significantly reduced the elevation of plasma asparatate aminotransferase levels, as well as anti-TNF and SOD treatment, compared with controls, but this reduction was not seen in the AG group. These results were confirmed by histological examination using hematoxylin-eosin and in situ terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) staining. Increases in tumor necrosis factor-alpha (TNF-alpha) mRNA expression in liver and in plasma TNF-alpha levels, which were observed in control mice, were not significantly reduced by PE or RA. PE and RA treatments also did not significantly diminish iNOS mRNA expression or plasma nitrate/nitrite levels. Nitrotyrosine and N(epsilon)-(hexanonyl)lysine (HEL) production, the residue of oxidative stress, was observed in livers from controls, but not in those mice pretreated with PE or RA. These results suggest that the liver protection of RA is due to the scavenging or reducing activities-superoxide or peroxynitirite rather than to inhibition of TNF-alpha production.     

紫苏、白苏的抑菌实验

用不同溶剂浸泡或处理紫苏后,进行抑菌实验,结果表明：１０％以上浓度的紫苏水浸液,１２．５％水煎液和乙醇提取液对白色念球菌、新型隐球菌以及红色毛癣菌、石膏样小孢子癣菌、絮状表皮癣菌有较好的抑菌效果。紫苏、皱紫共、白苏对真菌均有抑菌作用,但皱紫苏对绿脓杆菌、白苏对克柔氏念球菌、白色念球菌以及绿脓杆菌效果更为明显。     

Clinical effects of a hop water extract on Japanese cedar pollinosis during the pollen season: a double-blind, placebo-controlled trial

The clinical effects of an oral administration of a hop water extract (HWE) on the improvement of Japanese cedar pollinosis (JCPsis) symptoms were investigated. In a double-blind, placebo-controlled trial, 39 subjects took a drink containing either 100 mg of HWE or a placebo for 12 weeks during the pollen season. Nasal symptoms (sneezing attacks, nasal discharge, and nasal obstruction) were assessed from the subjects' diaries. A clinical examination and blood sampling were carried out before and 4, 8 and 12 weeks after the initiation of treatment. As a result, a significant difference was observed in the symptom score and in the symptom-medication score 10 weeks after the intervention in comparison with the placebo group. Improvements were observed in nasal swelling, nasal color, amount of nasal discharge, and characteristics of nasal discharge in the intervention group 12 weeks after the treatment. No significant eosinophil infiltration into the nasal discharge was apparent in the intervention group throughout the study period, although it was observed in the placebo group. These findings indicate that an oral administration of HWE may be effective in alleviating the allergic symptoms related to JCPsis.     

Clinical Effects of a Hop Water Extract on Japanese Cedar Pollinosis during the Pollen Season: A Double-Blind,Placebo-Controlled Trial

The clinical effects of an oral administration of a hop water extract (HWE) on the improvement of Japanese cedar pollinosis (JCPsis) symptoms were investigated. In a double-blind, placebo-controlled trial, 39 subjects took a drink containing either 100 mg of HWE or a placebo for 12 weeks during the pollen season. Nasal symptoms (sneezing attacks, nasal discharge, and nasal obstruction) were assessed from the subjects’ diaries. A clinical examination and blood sampling were carried out before and 4, 8 and 12 weeks after the initiation of treatment. As a result, a significant difference was observed in the symptom score and in the symptom-medication score 10 weeks after the intervention in comparison with the placebo group. Improvements were observed in nasal swelling, nasal color, amount of nasal discharge, and characteristics of nasal discharge in the intervention group 12 weeks after the treatment. No significant eosinophil infiltration into the nasal discharge was apparent in the intervention group throughout the study period, although it was observed in the placebo group. These findings indicate that an oral administration of HWE may be effective in alleviating the allergic symptoms related to JCPsis.     

紫苏属植物染色体数目和核型分析

通过对26份紫苏属材料进行染色体数目的观察,并对其1种(包括紫苏和白苏)及2变种材料各一份进行核型分析.结果显示,紫苏属植物染色体数均为2n=40,其核型公式分别为紫苏[P.frutescens(L.)Britton var.frutescens(zi-su)]2n=40=20m(2SAT) 10sm 10st;野生紫苏[P.frutescens var.purpurascens(Hayata)H.W.Li]2n=40=18m 14sm(2SAT) 8st;回回苏[P.frutescens var.crispa(Benth.)Deane ex Bailey]2n=40=10m(2SAT) 14sm 16st;白苏[P.frutescens(L.)Britton var.frutescens(bai-su)]2n=40-24m 12sm(2SAT) 4st.结果表明,紫苏属不同原(变)种植物的染色体数日相同,但在核型上存在一定的差异,其染色体核型为首次报道.     

温度和紫外照射对紫苏FAD7基因表达的影响

植物ω-3脂肪酸脱氢酶(ω-3FAD)基因是催化亚油酸转化为α-亚麻酸(ALA)的关键酶基因,通过调节该基因的表达,可以提高植物ALA的含量。为了研究温度和紫外照射对紫苏PfFAD7的影响,通过RTPCR方法分析了紫苏地上组织的特异性表达和温度、紫外胁迫下紫苏叶片和茎中PfFAD7基因的积累情况。分析表明,PfFAD7基因在紫苏全植株中均有表达,但在叶片中表达量最高,温度和紫外照射均影响PfFAD7基因的表达,低温可诱导PfFAD7基因的表达,而高温则抑制PfFAD7基因的表达;UV-B照射下,PfFAD7基因在叶片和茎中表达量均表现为先升高再降低。本试验对于紫苏ω-3脂肪酸脱氢酶的研究有利于高水平ALA的积累,更有利于紫苏资源的开发和利用,同时对于进一步了解不饱和脂肪酸的积累和代谢过程以及关键基因PfFAD7在此过程中的功能提供了依据。     

Increased carotenoid production in Xanthophyllomyces dendrorhous G276 using plant extracts

The red yeast Xanthophyllomyces dendrorhous (previously named Phaffia rhodozyma) produces astaxanthin pigment among many carotenoids. The mutant X. dendrorhous G276 was isolated by chemical mutagenesis. The mutant produced about 2.0 mg of carotenoid per g of yeast cell dry weight and 8.0 mg/L of carotenoid after 5 days batch culture with YM media; in comparison, the parent strain produced 0.66 mg/g of yeast cell dry weight and a carotenoid concentration of 4.5 mg/L. We characterized the utilization of carbon sources by the mutant strain and screened various edible plant extracts to enhance the carotenoid production. The addition of Perilla frutescens (final concentration, 5%) or Allium fistulosum extracts (final concentration, 1%) enhanced the pigment production to about 32 mg/L. In a batch fermentor, addition of Perilla frutescens extract reduced the cultivation time by two days compared to control (no extract), which usually required five-day incubation to fully produce astaxanthin. The results suggest that plant extracts such as Perilla frutescens can effectively enhance astaxanthin production.     

Comparative proteomics of nasal fluid in seasonal allergic rhinitis

A comparative proteomic approach was applied to examine nasal lavage fluid (NLF) from patients with seasonal allergic rhinitis (SAR, n = 6) and healthy subjects (controls, n = 5). NLF samples were taken both before allergy (pollen) season and during season, and proteins were analyzed by two-dimensional gel electrophoresis (2-DE) and matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) after tryptic cleavage. Twenty proteins were selected and quantified. During allergy season, the levels of six sialylated isoforms of PLUNC (palate lung nasal epithelial clone) were lower in SAR patients than controls, as were the levels of six isoforms of von Ebner's gland protein (VEGP), including a previously undescribed form with N-linked glycosylation, and of cystatin S. PLUNC is a new innate immunity protein and VEGP and cystatin S are two endogenous proteinase inhibitors. By contrast, the levels of an acidic form of alpha-1-antitrypsin were higher in SAR patients than controls. One previously unidentified NLF protein was found in all samples from the SAR patients during allergy season but not in any sample before allergy season: this protein was identified as eosinophil lysophospholipase (Charcot-Leyden crystal protein/galactin 10). MS/MS analysis of the N-terminus of the protein showed removal of Met and acetylation of Ser. Altogether, these findings illustrate the potential use of proteomics for identifying protein changes associated with allergic rhinitis and for revealing post-translational modifications of such new potential markers of allergic inflammation.     

Novel Antioxidants Isolated from Perilla frutescens Britton var. crispa (Thunb.)

Two novel antioxidants (vinyl caffeate and trans-p-menth-8-en-7-yl caffeate) and seven known antioxidants (3,4-dihydroxybenzaldehyde, methyl 3,4-dihydroxy-benzoate, methyl caffeate, 3′,4′,5,7-tetra-hydroxy-flavone, caffeic acid, 6,7-dihydroxycoumarin, and rosmarinic acid) were isolated from Perilla frutescens Britton var. crispa (Thunb.). The redox potentials of the novel isolated antioxidative compounds were comparable to those of known antioxidants. trans-p-menth-8-en-7-yl caffeate was effective to prevent the oxidative degradation of perillaldehyde in the essential oil of P. frutescens.     

Pharmacokinetics,Safety and Tolerability of Melissa officinalis Extract which Contained Rosmarinic Acid in Healthy Individuals: A Randomized Controlled Trial

The aim of this study was to evaluate the safety, tolerability and pharmacokinetics of single dose of Melissa officinalis extract which contained rosmarinic acid, including food-effects in healthy individuals. A total of eleven healthy individuals were randomly assigned to treatment arms in the two studies [Study 1 (fasted state) and Study 2 (fed state)]. Rosmarinic acid in serum was measured by a coulometric detection method using High-Performance Liquid Chromatography electrochemical detector. The serum concentration of total rosmarinic acid peaked at 1 hour after administration of Melissa officinalis extract containing 500mg rosmarinic acid in fasted state, with a maximum serum concentration 162.20 nmol/ L. The area under the curve for intact rosmarinic acid was calculated from the serum concentration-time profile to be 832.13 nmol • hour/ L. Food intake increases area under the curve and delayed time at which the maximum serum concentration. Rosmarinic acid supplementation did not affect liver, kidney, or blood cell function parameters. No adverse event was reported by any of the participants due to the study treatment. Single dose of Melissa officinalis extract containing 500 mg rosmarinic acid appears to be safe and tolerable in healthy individuals. Food intake increased the exposure of rosmarinic acid and delayed absorption of rosmarinic acid in healthy individuals.

Trial Registration

Trial Registration: UMIN-CTR UMIN000004997     

Effect of n-3 polyunsaturated fatty acid supplementation in patients with rheumatoid arthritis: a 16-week randomized,double-blind,placebo-controlled,parallel-design multicenter study in Korea

N-3 polyunsaturated fatty acids (PUFA) have anti-inflammatory effects and may be useful for the treatment of inflammatory diseases such as rheumatoid arthritis (RA).We examined the efficacy of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) supplementation on RA on top of standard anti-inflammatory treatment. Patients with RA were randomized into two groups in a double-blind, placebo-controlled, parallel-design multicenter study. One hundred nine patients received five capsules of either n-3 PUFA (2.090 g of EPA and 1.165 g of DHA) or high-oleic-acid sunflower oil for 16 weeks. Eighty-one patients completed the study, and no adverse effects were reported. Dietary intake did not change significantly during the study. There were significant increases in n-3 PUFA and EPA levels in erythrocytes in the n-3 PUFA group versus the placebo group, but decreases in n-6 PUFA, 18:2n6, 20:4n6 and 18:1n9 levels in the n-3 PUFA group versus the placebo group. N-3 PUFA supplementation had no significant effects on nonsteroidal anti-inflammatory drug (NSAID) requirements, clinical symptoms of RA or the concentration of cytokines, eicosanoids and bone turnover markers. However, n-3 PUFA supplementation significantly decreased NSAID requirements and leukotriene B₄ levels in patients who weighed more than 55 kg. Our results suggest that n-3 PUFA supplementation has no significant effect on RA but may decrease the requirement for NSAIDs in Korean patients with RA who weigh more than 55 kg.     

AsianPerilla crops and their weedy forms: Their cultivation, utilization and genetic relationships

The cultivation and utilization of two Perilla crops were surveyed in Asia. Perilla frutescens var. frutescens is essentially an oil crop and is now widely cultivated in China and Korea. Its seeds are also used as a flavor for traditional foods in Japan, Korea, China and Nepal. In Korea, leaves of var. frutescens are used as a fresh vegetable and for making pickles. Whereas P. frutescens var. crispa is a Chinese medicine and afresh vegetable in the Far East, it has almost disappeared in many parts of Asia. Cultivation of var. crispa is still continued in Japan and Vietnam. In particular, it is cultivated in a large scale for coloring pickles in the areas where a large amount of plum pickles are produced in Japan. In China and Korea, it remains only as a relict form. Weedy plants ofPerilla are found in Japan, Korea and China. We can classify them into two forms; one, which is closely related to var. frutescens, and the other, which is similar to var. crispa We foundP citriodora andR hirtella in Guandong and Jiangxi provinces of China, respectively. It is clear that they are not endemic to Japan. A phylogenetic tree of samples of two Perilla crops and their weedy forms based on RAPD markers revealed that the weedy forms similar to var. crispa and var. frutescens are genetically closely related to var. crispa and var. frutescens, respectively. Var. crispa and its closely related weedy form seem to be more primitive.     

Plasma extravasation through neuronal stimulation in human nasal mucosa in the setting of allergic rhinitis

Sanico, Alvin M., Satsuki Atsuta, David Proud, and AlkisTogias. Plasma extravasation through neuronal stimulation in humannasal mucosa in the setting of allergic rhinitis. J. Appl. Physiol. 84(2): 537-543, 1998.We havepreviously shown that capsaicin nasal challenge in subjects withallergic rhinitis produces a dose-dependent increase in the albumincontent of nasal lavage fluids. In the present set of studies, wedetermined whether this observation represents plasma extravasationthat is neuronally mediated. To evaluate whether glandular secretionscontribute to the albumin increase in nasal lavage fluids, volunteerswith allergic rhinitis were pretreated with atropine or placebo before capsaicin challenge. Atropine significantly reduced the volume ofreturned lavage fluids and their lysozyme content but increased theiralbumin and fibrinogen content. To assess the contribution of sensorynerve stimulation, subjects with allergic rhinitis were pretreated in asecond study with lidocaine or placebo before capsaicin challenge.Lidocaine significantly attenuated the capsaicin-induced increases inthe volume of nasal lavage fluids, as well as their lysozyme andalbumin content. To rule out the possibility of a direct effect oflidocaine on blood vessels rather than on nerves, healthy subjects werepretreated in a third study with lidocaine or placebo before bradykininnasal challenge. Lidocaine did not affect the bradykinin-inducedincrease in the albumin content of nasal fluids. We conclude that, inallergic rhinitis, high-dose capsaicin induces plasma extravasation inthe human nose and that this effect is neuronally mediated. Thisprovides more definitive evidence that neurogenic inflammation canoccur in vivo in the human upper airway.

     

1,2-Di-O-α-linolenoyl-3-O-β-galactosyl-sn-glycerol as a superoxide generation inhibitor from Perilla frutescens var. crispa

Using a superoxide (O(2)(-)) generation assay system with differentiated HL-60 cells, 1,2-di-O-α-linolenoyl-3-O-β-galactosyl-sn-glycerol (DLGG) was identified as an O(2)(-) generation inhibitor from Perilla frutescens var. crispa (a local variety, kida-chirimen shiso). DLGG suppressed the O(2)(-) level in a dose-dependent manner with an IC(50) value of 21 μM, comparable to those of rosmarinic acid (RoA, IC(50) = 29 μM) and caffeic acid (CA, IC(50) = 30 μM). While RoA and CA also dose-dependently inhibited O(2)(-) generation in a xanthine-xanthine oxidase system, DLGG had no effect in the same system. Thus DLGG appeared to decrease the O(2)(-) level in the HL-60 assay system by mechanisms different from those of RoA and CA, which appeared to act as O(2)(-) scavengers.     

云南境内紫苏种下变异的研究

选取26个性状,应用不加权算术平均配对法对来自云南各地的20份紫苏试材进行了数值分类学研究。结果表明:这20份紫苏试材可分成紫苏、回回苏、耳齿紫苏、野生紫苏和白苏5个变种;并对白苏变种的设立和耳齿紫苏变种的分布进行了探讨。     

Effects of glucan treatment on the Th1/Th2 balance in patients with allergic rhinitis: a double-blind placebo-controlled study

BACKGROUND: Allergic rhinitis (AR) is a disease characterized by IgE-mediated, allergic inflammation of the nasal mucosa. T helper (Th) 2 cells play an important role in the development of IgE-mediated diseases such as AR, with local overproduction of Th2 cytokines (IL-4, IL-5 and IL-13) at the site of allergic inflammation. Th1 cytokines (IL-12 and IFN-gamma) are known to suppress this Th2 immune response, aiding the treatment of these diseases. Beta-1,3-1,6-glucan (Glucan) is an immunomodulator stimulating particularly the antitumor response. An efficient antitumor stimulation can be achieved through a Th1-mediated immune response. OBJECTIVE: The aim of this study was to investigate the effects of Glucan on the immunopathogenic processes in the microenvironment to determine if it reverses the Th2-mediated immune response in AR to Th1-mediated response. METHODS: 24 Olea europea mono-sensitized patients with AR were randomized into Glucan and placebo groups. The Glucan group consisted of 12 patients who received Glucan treatment for 12 weeks, while the placebo group of 12 patients received placebo during the same period. A nasal provocation test (NPT) with Olea europea was performed at the beginning and end of treatment, and nasal lavage followed the positive NPT. IL-4, IL-5, IFN-gamma and IL-12 levels and the eosinophil count (%) were measured in nasal lavage fluid (NLF) samples. Simultaneously, peripheral blood eosinophil % values were measured. RESULTS: After treatment, IL-4 and IL-5 levels in NLF from the Glucan group were found to have decreased significantly (p = 0.027, p = 0.04; respectively), while IL-12 levels were found to have significantly increased (p = 0.008). However, IFN-gamma levels had not changed. On the other hand, none of the cytokine levels had changed significantly in the placebo group following treatment. Moreover, the percentage of eosinophils in the NLF was found to have decreased significantly after treatment in the Glucan group (p = 0.01), while that of the placebo group did not change. Peripheral blood percentage eosinophil levels had not changed significantly in any group. CONCLUSION: Th2-originated IL-4 and IL-5 levels responsible for the allergic inflammatory response in the microenvironment of patients with AR, are decreased with Glucan while levels of Th1-originated IL-12 are increased. Moreover, eosinophils, which are important effector cells of the inflammatory response, are decreased in the microenvironment. As a result, Glucan may have a role as an adjunct to standard treatment in patients with AR.     

No effect of folic acid supplementation on global DNA methylation in men and women with moderately elevated homocysteine

A global loss of cytosine methylation in DNA has been implicated in a wide range of diseases. There is growing evidence that modifications in DNA methylation can be brought about by altering the intake of methyl donors such as folate. We examined whether long-term daily supplementation with 0.8 mg of folic acid would increase global DNA methylation compared with placebo in individuals with elevated plasma homocysteine. We also investigated if these effects were modified by MTHFR C677T genotype. Two hundred sixteen participants out of 818 subjects who had participated in a randomized double-blind placebo-controlled trial were selected, pre-stratified on MTHFR C677T genotype and matched on age and smoking status. They were allocated to receive either folic acid (0.8 mg/d; n = 105) or placebo treatment (n = 111) for three years. Peripheral blood leukocyte DNA methylation and serum and erythrocyte folate were assessed. Global DNA methylation was measured using liquid chromatography-tandem mass spectrometry and expressed as a percentage of 5-methylcytosines versus the total number of cytosine. There was no difference in global DNA methylation between those randomized to folic acid and those in the placebo group (difference = 0.008, 95%CI = -0.05,0.07, P = 0.79). There was also no difference between treatment groups when we stratified for MTHFR C677T genotype (CC, n = 76; CT, n = 70; TT, n = 70), baseline erythrocyte folate status or baseline DNA methylation levels. In moderately hyperhomocysteinemic men and women, long-term folic acid supplementation does not increase global DNA methylation in peripheral blood leukocytes.ClinicalTrials.gov NCT00110604.     

Antimicrobial activity of perilla seed polyphenols against oral pathogenic bacteria

A perilla seed (Perilla frutescens Britton var. japonica Hara) extract was examined for its antimicrobial activity against oral cariogenic streptococci and periodontopathic Porphyromonas gingivalis. Luteolin, one of the components of perilla seed, showed the strongest antimicrobial effect among the phenolic compounds. According to our results, perilla seed may be the source of an antimicrobial agent that could prevent dental caries and periodontal diseases.