紫苏子镇咳、祛痰、平喘作用的药理研究

目的 对紫苏子及其炮制品的梯度溶媒提取物进行了镇咳、祛痰、平喘的对比性研究。方法 浓氨水喷雾法、毛细玻管法、喷雾致喘法。结果 1．紫苏子水提物、醇提物和醚提物均显示了程度不同的镇咳作用。2．紫苏子和炒紫苏子水提物的小剂量组均有良好的祛痰作用。3．对1％氯化乙酰胆碱诱导的豚鼠哮喘未发现其有平喘作用。但对用2％氯化乙酰胆碱和0．1％磷酸组胺的等量混合液诱喘的哮喘模型，炒紫苏子水提物和醚提物的小剂量组都显示出显著的平喘效果。结论 紫苏子具有一定的镇咳、祛痰和平喘作用，其镇咳成分较分散。平喘成分的水溶性大。存在于炒紫苏子的平喘成分其极性较分散，既存在于极性大的部分也存在于极性小的部分。     

An Intestinal Propulsion Promoting Substance from Perilla frutescens and Its Mechanism of Action

Perillaketone, a monoterpenoid component of the essential oil, was isolated from the leaf of PERILLA FRUTESCENS as an active principle of intestinal propulsion in mice. Pharmacological studies on the mechanism of action of this compound have suggested that the promotion of intestinal propulsion, which is accompanied with an increasing rate of the transport of contents through the intestinal canal, is largely due to stimulation of the motility of circular muscles of the intestine.     

Luteolin as an anti-inflammatory and anti-allergic constituent of Perilla frutescens

Oral administration of the perilla leaf extract (PLE) to mice inhibits inflammation, allergic response, and tumor necrosis factor-alpha production. We also found that PLE suppressed the tumor necrosis factor-alpha (TNF-alpha) production in vitro. Using the inhibitory activity of TNF-alpha production in vitro as the index for isolation, we searched the active constituents from PLE and isolated luteolin, rosmarinic acid and caffeic acid as active components. Among the isolated compounds, only luteolin showed in vivo activity: inhibition of serum tumor necrosis factor-alpha production, inhibition of arachidonic acid-induced ear edema, inhibition of 12-O-tetradecanoylphorbol-13-acetate-induced ear edema and inhibition of oxazolone-induced allergic edema. These results suggest that luteolin is a genuinely active constituent which is accountable for the oral effects of perilla.     

Genetic analysis of nothoapiol formation in Perilla frutescens.

A new essential oil chemotype of Perilla frutescens was found in that plant variety from Che-ju island in Korea. The steam-distilled oil of this plant was examined by GC-MS and 32 compounds were identified. Its principal constituents were beta-caryophyllene, dillapiol and nothoapiol. Also, crossing experiments were performed between the new chemotype and known chemotypes to clarify the genetic control of the production of nothoapiol. The gene Na, which promotes conversion from dillapiol to nothoapiol, was suggested, and was considered to be closely linked with the allele D.     

Metabolites of orally administered Perilla frutescens extract in rats and humans

As a part of our search for bioactive substances from the leaves of Perilla frutescens BRITTON var. acuta KUDO (Perillae Herba, Labiatae), the aqueous extract was orally administered to rats and humans, and metabolites in the urine, plasma, and/or bile were analyzed by a high-performance liquid chromatograph (HPLC) equipped with a photodiode array detector. When the extract was administered to rats, 10 metabolites, trans-caffeic acid-4-O-sulfate (1), trans-p-coumaric acid-4-O-sulfate (2), trans-ferulic acid-4-O-sulfate (3), trans-m-coumaric acid-3-O-sulfate (4), trans-caffeic acid (5), m-hydroxyphenylpropionic acid (6), trans-p-coumaric acid (7), trans-m-coumaric acid (8), luteolin (9), and apigenin (10) were detected in the urine, whereas four metabolites, scutellarein-6, 7-di-O-beta-glucuronide (11), apigenin-4'-O-sulfate-7-O-beta-glucuronide (12), apigenin-7-O-beta-glucuronide (13), and diosmetin-7-O-beta-glucuronide (14) were found in the bile. Compounds 1-8 and 11-14 were also found in the plasma. When the extract was given to humans, however, two metabolites, 1-O-(2,4,5-trimethoxycinnamoyl)-beta-glucuronic acid (15) and apigenin-4'-O-beta-glucuronide (16), were found in the urine and plasma. Thus, a species difference in the metabolism of the extract constituents was observed between rats and humans. Structures 1-16 were identified based on their chemical and spectral data.     

Cloning of the cytochrome P450 enzyme from Perilla frutescens involved in nothoapiole biosynthesis

Journal of Natural Medicines - Phenylpropanoid volatile components are found in various plants and are useful in medicines, health foods, and fragrances. While the pharmacological actions and...     

High-performance liquid chromatographic analysis of bioactive triterpenes in Perilla frutescens

Perilla frutescens (L.) Britt. (Lamiaceae), a famous traditional Chinese medicine, has been used for the treatment of various diseases. To evaluate the quality of P. frutescens, a simple, rapid and accurate high-performance liquid chromatography (HPLC) method was developed for the assessment of three bioactive triterpene acids: tormentic acid (TA), oleanolic acid (OA) and ursolic acid (UA). The HPLC system used an Spherisob octadecylsilyl silica (ODS) column with acetonitrile and aqueous H₃PO₄ as the mobile phase and detection at 206 nm. The method was precise with relative standard deviations for these three constituents that ranged between 0.6–1.5% (intraday) and 0.7–2.6% (interday). The content of these three phytochemicals in the leaves of P. frutescens growing at eight different locations of China was determined to establish the effectiveness of the method.     

Suppressive effects of Perilla frutescens on mesangioproliferative glomerulonephritis in rats

Leaves of Perilla frutescens var. crispa DECNE. (perilla, Labiatae) are used as a garnishing vegetable in East Asian countries as well as an herbal medicine prescribed in Kampo medicines such as Saiboku-to. A previous in vitro study revealed that a decoction of perilla leaves inhibits the proliferation of murine-cultured mesangial cells. In the present study, we evaluated the in vivo anti-proliferative effects of a perilla decoction using rat mesangio-proliferative glomerulonephritis induced by an intravenous injection of rabbit anti-rat thymocyte serum (ATS). Leaves of perilla were boiled, and the decoction was orally administered to the rats as drinking water at doses of 100 and 500 mg/kg/d from the day of ATS-injection (day 0) to day 8, when rats were sacrificed. In the histological evaluation, the total number of glomerular cells, proliferating cell nuclear antigen (PCNA) positive cells, and macrophage/monocyte antigen-positive cells in the glomerulus, was significantly decreased in perilla-treated rats. A significantly lower level of proliferation was induced by the serum of the perilla-treated rats than by that of the controls. These results suggest that the perilla decoction suppresses the proliferation of mesangial cells in vivo by an inhibition of the glomerular infiltration of macrophage/monocytes and of the production of circulating growth factors.     

Intoxication of cattle by Perilla frutescens (purple mint)

Perilla frutescens or purple mint has been associated with atypical interstitial pneumonia (AIP) for a quarter of a century. The amount and the stage of the plant required to produce AIP have been much debated. A field case in which catastrophic loses occurred in cattle ingesting hay containing purple mint showed that more than the green plants have the capability of producing atypical interstitial pneumonia. In this study, Perilla frutescens produced atypical interstitial pneumonia in three of five calves to which it was given. The amount required to produce the syndrome ranged from 2.3 to 15.5 kg of green seed stage mint and 11.8 kg of mint hay. The toxic syndromes were similar in signs, but quite different in duration. Necropsy examinations showed varied amounts of pulmonary emphysema and edema. Two of the three affected animals' lungs histologically displayed a marked proliferation of Type II pneumocytes. The flowering or seed parts of perilla mint were found by high pressure liquid chromatographic analysis to contain the highest concentration of perilla ketone, considered the most toxic agent involved. This stage of plant growth was also shown to be the most toxic in our calf feeding trial. Calves fed the flowering plant developed the toxic syndrome while those fed earlier plants (collected before seed stage) and late plants (collected after frost) remained asymptomatic. The time of year when perilla reaches the seed stage often corresponds to periods when pasture grass is scarce forcing cattle to consume plants not normally eaten when ample desirable forage is available.(ABSTRACT TRUNCATED AT 250 WORDS)     

Sedative effects of inhaled Perilla frutescens essential oils on mice

Journal of Natural Medicines - Perilla frutescens is an ingredient for cooking and for Japanese traditional medicine formulations. Essential oils extracted from P. frutescens are classified...     

Genetic Control of Peltate Glandular Trichome Formation in Perilla frutescens

The inheritance of peltate glandular trichome (PGT) formation in PERILLA FRUTESCENS leaves was investigated by crossing a normal strain with a variant strain with sparse PGT formation. The F (1) and F (2) data suggested that the sparse trichome is a dominant character that is controlled by a major gene and a few modifying genes which specifically inhibit the development of trichomes of the peltate type. Tracer experiments using [ (14)C]-sucrose showed that the quantity of essential oil components synthesized in the leaf was positively correlated with the number of PGTs, suggesting that the PGT is the main site for the biosynthesis of essential oil.     

Rosmarinic acid in Perilla frutescens and perilla herb analyzed by HPLC

High-quality perilla leaves are defined as those having purple upper and lower surfaces and a pleasant smell. The Japanese Pharmacopoeia specifies the content of essential oils in perilla leaves but not the content of rosmarinic acid. Rosmarinic acid is a common component of Labiatae plants such as shiso (Perilla frutescens Britton var. crispa W. Deane). Rosmarinic acid has been reported to exhibit anti-inflammatory and anti-oxidant activity but the factors affecting the content of rosmarinic acid in plants remain unknown. This study describes a simple and reproducible method for quantifying rosmarinic acid. We elucidated the main causes for the different rosmarinic acid contents of plants by examining various samples of perilla using the proposed method. Significant differences in rosmarinic acid content between varieties and cultivators were observed. The rosmarinic acid content was higher in green perilla compared with red perilla, in wild species compared with cultivated species, and in plants cultivated in outdoor nurseries compared with in indoor nurseries. The proposed quantitative method was used to examine the rosmarinic acid content in a Kampo formula, Hangekobokuto, and was found to be higher in decoctions prepared using the Kouge method compared with the typical preparation method. We examined the chlorophyll and caffeic acid contents of several samples and their relationship with the rosmarinic acid content.     

Antidepressant-like effects of apigenin and 2,4,5-trimethoxycinnamic acid from Perilla frutescens in the forced swimming test

We studied the effects of apigenin and 2,4,5-trimethoxycinnamic acid (TMCA) on the behavioral despair test (forced swimming test), and the central noradrenergic, dopaminergic and serotonergic activities in mice. Apigenin at intraperitoneal doses of 12.5 and 25 mg/kg significantly decreased the duration of immobility in the forced swimming test in mice. At 100 mg/kg, the duration of immobility was returned to the control level in the test. On the other hand, TMCA treatment (25-200 mg/kg, i.p.) failed to significantly alter the duration of immobility. Based on the behavioral data, we examined changes in the monoamine turnover in mice having been subjected to forced swimming for 40 min. The monoamine turnover was measured in seven brain regions. Forced swimming exposure induced a significant decrease in dihydroxyphenylacetic acid (DOPAC)/dopamine (DA) in the striatum and amygdala and in 5-hydroxyindoleacetic acid (5-HIAA)/5-hydroxytriptamine (5-HT) in the hypothalamus, and a significant increase in DOPAC/DA in the thalamus and hypothalamus and in 3-methoxy-4-hydroxyphenylethyleneglycol (MHPG)/norepinephrine (NE) in the amygdala, frontal cortex, hypothalamus, and midbrain. Apigenin (25 mg/kg) treatment produced attenuation of forced swim test-induced decrease of DA turnover in the amygdala and increase of DA turnover in the hypothalamus. Furthermore, intraperitoneal administration of haloperidol (0.2 mg/kg), a dopamine D(2) antagonist, blocked the apigenin (25 mg/kg)-induced decrease in immobility in the forced swimming test. These behavioral and biochemical results indicate the antidepressant properties of apigenin, which may be mediated by the dopaminergic mechanisms in the mouse brain.     

Bioconversion of essential oil components of Perilla frutescens by Saccharomyces cerevisiae

Journal of Natural Medicines - The essential oil of perilla (Perilla frutescens) contains volatile low molecular weight compounds such as monoterpenes and phenylpropenes. The composition of the...     

炒紫苏子标准汤剂的质量标准研究

目的:制备炒紫苏子标准汤剂,并进行质量标准研究。方法:依照标准汤剂的制备要求,制备17批炒紫苏子标准汤剂,计算出膏率。采用高效液相色谱(HPLC)法对炒紫苏子标准汤剂中迷迭香酸进行定量分析[色谱柱为Agilent 5 TC-C18(2),流动相为甲醇-0.1%甲酸溶液(40∶60,V/V),检测波长为330 nm,流速为1.0 mL/min,柱温为30℃,进样量为5μL],并计算其转移率。建立17批炒紫苏子标准汤剂的HPLC指纹图谱,利用中药色谱指纹图谱相似度评价系统(2012版)软件对指纹图谱进行分析,并通过对比共有峰的保留时间对色谱峰进行指认。结果:17批炒紫苏子标准汤剂的出膏率为5.55%～9.75%;迷迭香酸的含量为0.44%～1.58%,平均含量为1.08%;迷迭香酸的转移率为18.31%～34.32%,平均转移率为25.42%。在17批炒紫苏子标准汤剂HPLC指纹图谱中,共确定了11个共有峰,相似度均高于0.95;并指认出了其中5个共有色谱峰,分别为咖啡酸(峰3)、木犀草苷(峰5)、迷迭香酸(峰8)、木犀草素(峰9)和芹菜素(峰10)。结论:建立了炒紫苏子标准汤剂的质量控制方法,可为炒紫苏子配方颗粒及相关制剂质量标准的制订提供参考。     

超临界CO_2萃取紫苏叶挥发油及其成分分析

目的:采用超临界流体萃取技术提取紫苏叶挥发油,并用GC-MS对挥发油进行化学成分分析。方法:通过单因素实验和正交实验法确定超临界CO2流体萃取紫苏叶挥发油的最佳条件,考察萃取压力、温度、动态萃取时间及CO2流量对挥发油得率的影响;利用GC-MS分析最佳萃取条件下所得挥发油的化学成分,面积归一化法测定其百分含量。结果:紫苏叶超临界CO2萃取最佳条件为:萃取压力20 MPa,温度35℃,萃取时间150 min,CO2流量10 kg.h-1;挥发油的得率3.2%,从中鉴定出了16个化合物,其含量占出峰物质总量的97.68%。结论:紫苏叶挥发油中富含醚、萜类化合物、酯、酮和醇等组分。     

In vitro and in vivo effects of macrophage-stimulatory polysaccharide from leaves of Perilla frutescens var. crispa

The crude polysaccharide (PFB-1) was isolated from the leaves of Perilla frutescens var. crispa by the sequential procedures with hot-water extraction, methanol reflux, and ethanol precipitation. It was further purified by anion column chromatography in order to obtain the partially purified polysaccharide (PFB-1-0). In the presence of PFB-1-0, strong cellular lysosomal enzyme activity of murine peritoneal macrophages was observed in vitro. Compared to bacterial lipopolysaccharide (LPS), its activity was relatively high. The in vitro phagocytic activity was enhanced by PFB-1-0 as the similar pattern in both gram-negative bacteria, E. coli, and gram-positive bacteria, S. aureus with a time-dependent manner. We also investigated the production of several mediators by murine peritoneal macrophages upon stimulation with PFB-1 (in vivo) or PFB-1-0 (in vitro). The levels of nitric oxide (NO) and tumor necrosis factor (TNF)-alpha were increased in the presence of PFB-1-0 in vitro. The PFB-1 stimulated the production of interleukin (IL)-6 and granulocyte-macrophage colony-stimulating factor (GM-CSF) in vivo. Results suggest that the polysaccharide from P. frutescens var. crispa represents an immunopotentiator and biological response modifiers in vitro and in vivo levels.     

Molecular cloning, functional expression and characterization of d-limonene synthase from Schizonepeta tenuifolia

Limonene is one of the most simple cyclic monoterpenes, and two enantiomers, d- and l-limonene occur due to the chiral carbon at 4-position. Cyclization of GPP into limonene is catalyzed by the limonene synthase, and some l-limonene synthase cDNAs have already been cloned from several species of plants, mainly from Labiatae family. However, the d-limonene synthase gene has not yet been obtained, therefore, no information is available on the molecular mechanism of stereochemical regulation in limonene formation. To resolve this, we cloned the d-limonene synthase gene (dLMS) from Schizonepeta tenuifolia (Labiatae) by a reverse genetic approach, and we found that both d- and l-limonene synthase share similar features such as a transit peptide, an arginine rich domain, and a metal cation binding site in their structures. Here, we report on the cloning of dLMS, and the putative stereochemical regulation mechanism is discussed based on the comparison of the deduced amino acid sequence of dLMS with those of known l-limonene synthases.     

Molecular cloning,functional expression and characterization of d-limonene synthase from Agastache rugosa

We cloned the gene of d-limonene synthase (ArLMS) from Agastache rugosa (Labiatae). The function of ArLMS was elucidated by the preparation of recombinant protein and subsequent enzyme assay. ArLMS consisted of 2077 nucleotides including 1839 bp of coding sequence that encodes a protein of 613 amino acids. This protein has a 60 kDa molecular weight, which is identical to that of d-limonene synthase from Schizonepeta tenuifolia (Labiatae). The deduced amino acid sequence of ArLMS shows high homology with the known d- and l-limonene synthases from Labiatae plants. Here, we discussed the amino acid residues responsible for the stereochemical regulation in limonene biosynthesis.     

Determination of Final Steps in Biosyntheses of Essential Oil Components in Perilla frutescens

Biosynthetic pathways of the essential oil components in PERILLA FRUTESCENS were examined by tracer experiments using [ (14)C]-labeled sucrose on two chemotypes: perillaketone (PK) and phenylpropanoid (PP-EM) types. The results of the experiments indicated that perillaketone is not converted from isoegomaketone but is synthesized directly from the hypothetical precursor egomaketone. On the other hand, it was shown that elemicin is not synthesized via myristicin but from the possible common precursor, methyleugenol, in the plant having the dominant gene E. These results validate the hypothetical pathways proposed by Hegnauer and Fujita.