Lycopus Lucidus Turcz Research Articles

Lactobacillus paracasei HY7015 and Lycopus lucidus Turcz. Extract Promotes Human Dermal Papilla Cell Cytoprotective Effect and Hair Regrowth Rate in C57BL/6 Mice

Hair loss is a disease that requires accurate diagnosis and type-specific medical treatment. Many hair loss treatments have some side effects, such as hormone-related effects, so there is a need for safe and effective hair loss treatment. In this study, we investigated the effects of Lactobacillus paracasei HY7015 (HY7015) and Lycopus lucidus Turcz. (LT) extract on hair regrowth and protection. In vitro experiments were conducted to assess the effects of HY7015 and/or LT extract on human follicle dermal papilla cells (HFDPC) of cytoprotective functions such as proliferations, antioxidants, anti-inflammatory, and growth factor expressions. In animal experiments, we investigated hair regrowth rate, hair follicle formation and secretion of growth factors in telogenic C57BL/6 mice. We confirmed the cytoprotective effects of HY7015 and LT through regulations of proliferation, SOD and IL-1β in HFDPC. In mouse experiments, oral administration of HY7015 and LT promoted hair regrowth as well as hair follicle maturation in the dermal skin of C57BL/6 mice, and upregulated VEGF and IGF-1 growth factor levels in mouse serum. In summary, our data demonstrate that ingestions of HY7015 and LT can promote hair regrowth by enhancing cytoprotective effects and expressions of growth factors.

Effect of Fractions from Lycopus lucidus Turcz. Leaves on Genomic DNA Oxidation and Matrix Metalloproteinase Activity.

We investigated the inhibitory effects of fractions from Lycopus lucidus Turcz. leaves on genomic DNA oxidation, Nitric Oxide (NO) production, and Matrix Metalloproteinase (MMP) activity. Oxidative damage of genomic DNA was detected after Fenton reaction with H2O2 using DNA electrophoresis. Western blotting was performed to compare the expression levels of MMP-2 in phorbol 12-myristate 13-acetate (PMA)-induced HT-1080 cells. Lipopolysacchride (LPS)-induced NO production in RAW 264.7 cells was measured using Griess reagent. All fractions (n-Hexane, 85% aq. MeOH, n-BuOH, and water fractions) from the leaves of L. lucidus Turcz. significantly inhibited intracellular production of reactive oxygen species (ROS) (p<0.05). Particularly, 85% aq. MeOH and n-BuOH fractions showed higher ROS inhibitory activity than the other fractions. n-Hexane, 85% aq. MeOH, n-BuOH and water (0.05 mg/mL) fractions significantly inhibited oxidative DNA damage by 57.97%, 68.48%, 58.97%, and 68.39%, respectively (p <0.05). Treatment of RAW 264.7 cells with each fraction reduced LPS-induced NO production in a dose-dependent manner (p<0.05). n-Hexane and 85% aq. MeOH fractions notably reduced MMP-2 secretion levels in the culture supernatants from HT-1080 cells. Overall, these results indicated that L. lucidus Turcz. leaves can be exploited as plant based sources of antioxidants in the pharmaceutical, cosmetic, nutraceutical, and food industries.

Lycopus lucidus Turcz Water Extract Ameliorates the Metabolic Disorder by Up-Regulated Major Urinary Protein Expression in High-Fat Diet-Induced Obesity.

Despite a century of research on obesity, metabolic disorders and their complications, including dyslipidemia, insulin resistance, and fatty liver disease remain a serious global health problem. Lycopus lucidus Turcz (LT) is a traditional medicine used for its anti-inflammatory properties that has not been evaluated for its efficacy in improving obesity. In this study, mice were fed a normal diet (n = 10) or obesity was induced with a high-fat diet (HFD, n = 20, 60% kcal from fat) for 4 weeks. The HFD mice were then divided into two groups, one of which received LT supplementation with water extract for 13 weeks [HFD (n = 10) or HFD with LT water extract (n = 10, 1.5%)]. LT reduced body and adipose tissue weight by elevating energy expenditure by increasing fatty oxidation in epididymal white adipose tissue (eWAT) and muscle. LT ameliorated dyslipidemia and hepatic steatosis by restricting lipogenesis. Additionally, LT normalized the impaired glucose homeostasis by diet-induced obesity to improve pancreatic islet dysfunction with increasing hepatic major urinary protein expression. Moreover, LT attenuated the inflammation and collagen accumulation in the liver and eWAT. In conclusion, these results suggest that LT can treat obesity-related metabolic disorders such as adiposity, dyslipidemia, hepatic steatosis, insulin resistance, and inflammation.

Lycopus lucidus Turcz ameliorates DNCB‑induced atopic dermatitis in BALB/c mice.

Atopic dermatitis (AD) is a chronic inflammatory allergic skin disease, characterized by pruritic and eczematous skin lesions. Lycopus lucidus Turcz (LLT) is a perennial herb that has been reported to have various biological properties, including effects on blood circulation, as well as anti-inflammatory, antioxidant, anti-vascular inflammation and wound-healing effects. However, whether LLT improves dermatitis and the underlying mechanisms has yet to be determined. The aim of the present study was to determine whether LLT can improve 2,4-dinitrochlorobenzene (DNCB)-induced dermatitis and to verify the inhibitory effect of LLT on the expression of chemokines and pro-inflammatory cytokines in the HaCaT immortalized keratinocyte cell line. In addition, the anti-inflammatory function of LLT in RAW264.7 mouse macrophages was investigated. In the DNCB-induced AD mouse model, LLT inhibited infiltration by mast cells, eosinophils and CD8+ cells in the dorsal skin tissue of AD mice, and suppressed the expression of IgE and IL-6 in serum. In addition, LLT inhibited the phosphorylation of ERK and JNK, as well as NF-κB in skin tissue. In the HaCaT cell model induced by TNF-α/IFN-γ, LLT inhibited the expression of thymus and activation-regulated chemokine, granulocyte-macrophage colony-stimulating factor, monocyte chemoattractant protein-1, TNF-α and IL-1β, whilst inhibiting the phosphorylation of NF-κB. In addition, in the lipopolysaccharide-induced RAW 264.7 cell inflammation model, LLT inhibited the expression of TNF-α and IFN-γ, the nuclear translocation of NF-κB and the phosphorylation of ERK and JNK. These results suggested that LLT may be a promising candidate for the treatment of inflammatory dermatitis.

Lycopus lucidus Turcz Exerts Neuroprotective Effects Against H2O2-Induced Neuroinflammation by Inhibiting NLRP3 Inflammasome Activation in Cortical Neurons.

PurposeLycopus lucidus Turcz (LLT) is a potent traditional medicinal herb that exerts therapeutic effects, regulating inflammatory disorders. However, the precise mechanisms by which LLT plays a potent role as an anti-inflammatory agent are still unknown, and in particular, the effects of LLT on cortical neurons and related mechanisms of neuroinflammation have not been studied. The NLRP3 inflammasome pathway is one of the most well known as an important driver of inflammation. We therefore hypothesized that LLT, as an effective anti-inflammatory agent, might have neurotherapeutic potential by inhibiting the NLRP3 inflammasome pathway in cortical neurons.Materials and MethodsPrimary cortical neurons were isolated from the embryonic rat cerebral cortex, and H2O2 was used to stimulate neuron damage in vitro. After treatment with LLT at three concentrations (10, 25, and 50 µg/mL), the expression of iNOS, NLRP3, ASC, caspase-1, IL-1β, IL-18, IL-6, and IL-10 was determined by immunocytochemistry, qPCR, and ELISA. Neuron apoptosis was also evaluated using Annexin V-FITC/PI double staining FACS analysis. Neural regeneration-related factors (BDNF, NGF, synaptophysin, NT3, AKT, and mTOR) were analyzed by immunocytochemistry and qPCR.ResultsLLT effectively protected cultured rat cortical neurons from H2O2-induced neuronal injury by significantly inhibiting NLRP3 inflammasome activation. In addition, it significantly reduced caspase-1 activation, which is induced by inflammasome formation and regulated the secretion of IL-1β/IL-18. We demonstrated that LLT enhances axonal elongation and synaptic connectivity upon H2O2-induced neuronal injury in rat primary cortical neurons.ConclusionIt was first demonstrated in vitro that LLT suppresses NLRP3 inflammasome activation, attenuates inflammation and apoptosis, and consequently promotes neuroprotection and the stimulation of neuron repair, suggesting that it is a promising therapeutic for neurological diseases.

The protective effects of lycopus lucidus turcz in diabetic retinopathy and its possible mechanisms

The aim of the present study was to investigate the effect of Lycopus lucidus Turcz (LT) on diabetic retinopathy (DR) and its underlying mechanisms. SD rats and human retinal microvascular endothelial cells (HRECs) were applied for establishment DR model. HE and TUNEL staining were used to evaluate the pathological changes and apoptosis of retinal ganglion cells. Additionally, retinal vessels were detected by immunofluorescence staining with CD31 and VEGF. The function of BRB was observed using Evans blue. Moreover, the oxidative stress, inflammation and angiogenesis associated factors were measured respectively. The expression of p38-MAPK/NF-κB signalling proteins were detected by Western blot. The results demonstrated that pathological changes and retinal optic disc cells apoptosis in retinas of diabetic rats, both of which were reduced in the LT-treated group. And LT treatment attenuated the levels of oxidative stress, inflammation and angiogenesis factors. Importantly, the expression levels of p-p38, p-ERK, p-JNK and NF-κB were decreased. After treatment with TNF-α combined with LT, the levels of inflammatory factors were decreased but higher than the negative control. Taken together, the results suggested that LT treatment is of therapeutic benefit by ameliorating oxidative stress, inflammation and angiogenesis of DR via p38-MAPK/NF-κB signaling pathway.

Protective effect of free phenolics from Lycopus lucidus Turcz. root on carbon tetrachloride-induced liver injury in vivo and in vitro.

Protective effect of free phenolics from Lycopus lucidus Turcz. root (FPLR) on CCl4-induced hepatotoxicity in vivo and in vitro was first evaluated. Oral administration of FPLR (100 mg/kg bw) to mice significantly reduced the CCl4-induced elevation of serum alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, triacylglycerols, total cholesterol, and total bilirubin. FPLR also increased the hepatic GSH contents and antioxidant enzyme activities of SOD and CAT and decreased the hepatic MDA level. Histopathological examinations further confirmed that the FPLR could protect the liver from CCl4-induced damage. Further research indicated that FPLR prevented the DNA fragmentation caused by CCl4 based on TUNEL assay. Moreover, immunohistochemistry staining demonstrated that pretreatment with FPLR significantly inhibited the elevation of hepatic TNF-α, IL-6, IL-8, iNOS, COX-2, and Caspase-3 in CCl4-treated mice. In vitro experiments showed that FPLR remarkably reduced BRL hepatocyte apoptosis and damage caused by CCl4 treatment. These findings indicate that FPLR could be developed as a functional food or medication for therapeutic purpose and prevention of hepatic injury.

Lycopus lucidus Turcz. ex Benth. Attenuates free fatty acid-induced steatosis in HepG2 cells and non-alcoholic fatty liver disease in high-fat diet-induced obese mice

BackgroundNon-alcoholic fatty liver disease (NAFLD) is closely related to metabolic diseases such as obesity and insulin resistance. PurposeWe studied whether an ethanol extract of Lycopus lucidus Turcz. ex Benth (LLE) exhibited effects on lipid metabolism in NAFLD. Study designAn in vitro modelwas established by treatment of HepG2 cells with a 1 mM free fatty acid (FFA) mixture (oleic acid/palmitic acid, 2:1). C57BL/6 mice were fed a high-fat diet (HFD; 60 kcal% fat) for 14 weeks to induce obesity and were treated with or without LLE (100 or 200 mg/kg daily by oral gavage). MethodsHepG2 cells were exposed to 1 mM FFA, with or without LLE (250 – 1000 mg/ml). Intracellular lipid contents were measured by Oil Red O staining and a Nile Red assay. The body weight, relative liver weight, hepatic lipids, triglycerides (TGs), and total cholesterol (TC) were measured in the mice. Serum alanine aminotransferase (ALT), TG, TC, glucose, insulin, leptin, and tumor necrosis factor-alpha (TNF-α) levels were determined by biochemical or enzyme-linked immunosorbent assays. Histologic analysis was performed in the liver. Western blotting and quantitative real-time polymerase chain reaction were used to analyze the expression of key enzymes of hepatic lipid metabolism. ResultsLLE significantly decreased the intracellular lipid accumulation in FFA-treated HepG2 cells. LLE not only remarkably decreased the expression of lipogenesis genes but also increased β-oxidation in FFA-induced HepG2 cells. In the in vivo study, LLE treatment significantly decreased the body weight, relative liver weight, serum ALT, TC, and low-density lipoprotein cholesterol, as well as the serum glucose, insulin, leptin, and TNF-α levels in HFD-fed mice. The hepatic TG and TC contents were significantly reduced in the LLE-treated groups. Western blot analysis showed that the expression of sterol-regulatory element-binding protein 1 decreased, while that of phosphorylated AMP-activated protein kinase and peroxisome proliferator-activated receptor α increased in the LLE-treated mice. ConclusionThese results suggest that LLE may exert protective effects against NAFLD-related obesity and metabolic disease.

Ethanol Extract of Lycopus lucidus Turcz. ex Benth Inhibits Metastasis by Downregulation of Runx-2 in Mouse Colon Cancer Cells.

Lycopus lucidus Turcz. ex Benth (LT) has been broadly used as a traditional medicinal herb in Asia including Korea, China, and Japan due to its noted ability to promote blood circulation and remove blood stasis. However, its anticancer mechanism is not understood. This study aims to elucidate the effects of ethanol extracts of LT (ELT) relative to the role of Runt-related transcription factor- (Runx-) 2 in the invasive and metastatic potentials of mouse colon cancer to determine the underlying mechanisms involved. ELT was evaluated for the antimetastasis activity using CT-26 colon cancer using wound healing, transwell matrigel, and western blot analysis. We used Runx-2-specific siRNA to further determine the relationship between Runx-2 and matrix metalloprotease- (MMP-) 9 in the migration and invasion of CT-26 cells. Runx-2 was first demonstrated to be a transcription factor that plays a remarkable role in diverse biological processes of chondrocytes and osteoblasts, but recently, Runx-2 has been reported to be associated with the progression of certain human cancers. ELT was not altered in its effects on growth inhibition. However, ELT significantly inhibited wound closure and cell invasion in a dose-dependent manner. ELT decreased the metastasis by regulating the activity of MMP-9 and Runx-2 at the translational levels. Our results demonstrate that ELT decreases metastasis by inhibiting the Runx-2–MMP-9 axis. We suggest that it can be used as a novel agent in therapeutic strategies for combating colon cancer.

쉽싸리 부위별 물추출물의 항산화 활성

Objectives : This study was performed to investigate the antioxidant activity of water extracts from Lycopus lucidus Turcz. ex Benth. leaves, stems and roots at the <TEX>$100{\mu}g/m{\ell}$</TEX> concentration. Methods : The different part of Lycopus lucidus Turcz. ex Benth. extract was prepared using water. The antioxidant activities of polyphenol contents, total flavonoid contents, DPPH(1,1-diphenyl-2-picrylhydrazyl) free-radical scavenging activity, SOD like activity, hydroxyl radical, ABTS(2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid), <TEX>$Fe^{2+}$</TEX> chelating, and nitrite scavenging activity. Results : The total polyphenol and total flavonoid content of leaves were the highest at <TEX>$221.85{\mu}g/mg$</TEX> and <TEX>$794.13{\mu}g/mg$</TEX>, respectively. Electron donating ability was the 79.68% in the water extract from leaves. The ABTS radical scavenging activity of the hot extracts, leaves <TEX>${\gg}$</TEX> roots > stems was higher in the order. It was shown the highest at 94.53% in the water extract from leaves, which showed a value equal to 94.7% of ascorbic acid. The hydroxyl radical scavenging activity was the highest at 8.07% in the water extract from leaves. SOD like activity and <TEX>$Fe^{2+}$</TEX>chelating activity were leaves of 12.3% and 27%, respectively, which were much higher than those of any other parts. The nitrite scavenging ability of extracts was increased at pH 2.5, and those was the highest in leaves of 83.03%. Its were more than twice the 41.61% of BHT. Conclusion : The results suggest that Lycopus lucidus Turcz. ex Benth. can be used as nutraceutical foods and natural antioxidant.

Chemical identification and quality evaluation of Lycopus lucidus Turcz by UHPLC-Q-TOF-MS and HPLC-MS/MS and hierarchical clustering analysis.

Lycopus lucidus Turcz has been used as a traditional phytomedicine for menstrual disorder, amenorrhea, menstrual cramps, inflammation and cardiovascular diseases. However, there is not enough information about identification and quantification for the chemical constituents of L. lucidus Turcz. In this work, a simple, rapid and sensitive UHPLC-Q-TOF-MS method was developed for characterization and identification of the phytochemical compositions in L. lucidus Turcz in negative ion mode. A total of 37 compounds, including 15 phenolic acids, 12 flavonoids, three triterpenoids and seven organic acids were tentatively characterized and identified by means of the retention time, accurate mass and characteristic fragment ions. Thirteen compounds were reported for the first time in L. lucidus Turcz. Among of them, 11 compounds were further quantified by multiple reactions monitoring. The results showed good performance with respect to linearity (r >0.9959), repeatability (RSD< 2.6%), intra- and inter-day precision (RSD< 3.2%), recovery (93.1-104.9%), and lower limit of quantification (5-50 ng/mL). Subsequently, the results were analyzed and classified by hierarchical cluster analysis. The research could be applied for identification and quality evaluation for L. lucidus Turcz.

Screening and identification of the metabolites in rat urine and feces after oral administration of Lycopus lucidus Turcz extract by UHPLC-Q-TOF-MS mass spectrometry.

Lycopus lucidus Turcz has been used as a kind of edible and medicinal material in eastern Asian countries. It has various bioactivities, including treatment of menstrual disorder, amenorrhea, menstrual cramps, inflammation, and cardiovascular diseases. However, the in vivo metabolism of L. lucidus Turcz extract is still not well described. In this study, L. lucidus Turcz extracts were administered to rats. Urine and fecal samples were collected at the difference periods (0-12h, 12-24h, and 24-36h). Ultra-high performance liquid chromatography coupled with electrospray ionization tandem quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) method was developed to characterize and identify the metabolites. A total of 17 metabolites in feces and 19 metabolites in urine were tentatively identified by means of accurate mass and characteristic fragment ions. The results show that glucuronidation and sulfation are the major metabolic reactions. This study is the first reported analysis and characterization of the metabolites and the proposed metabolic pathways of bioactive components might provide further understanding of the metabolic fate of the chemical constituents after oral administration of L. lucidus Turcz extract in rats.

Advance researches of health food—Lycopus lucidus Turcz

On the basis of summary and analysis of recent literature about the edible and medicinal properties of Lycopus lucidus Turcz,its nutrient content,chemical composition and the function of active ingredients were introduced in this paper.It showed that Lycopus lucidus Turcz was a kind of health food raw materials with huge potential for its rich nutrient and active ingredients.

Hydrophilic interaction liquid chromatography for the separation, purification, and quantification of raffinose family oligosaccharides from Lycopus lucidus Turcz.

A systematic strategy based on hydrophilic interaction liquid chromatography was developed for the separation, purification and quantification of raffinose family oligosaccharides from Lycopus lucidus Turcz. Methods with enough hydrophilicity and selectivity were utilized to resolve the problems encountered in the separation of oligosaccharides such as low retention, low resolution and poor solubility. The raffinose family oligosaccharides in L. lucidus Turcz. were isolated using solid-phase extraction followed by hydrophilic interaction liquid chromatography at semi-preparative scale to obtain standards of stachyose, verbascose and ajugose. Utilizing the obtained oligosaccharides as standards, a quantitative determination method was developed, validated and applied for the content determination of raffinose family oligosaccharides both in the aerial and root parts of L. lucidus Turcz. There were no oligosaccharides in the aerial parts, while in the root parts, the total content was 686.5 mg/g with the average distribution: raffinose 66.5 mg/g, stachyose 289.0 mg/g, verbascose 212.4 mg/g, and ajugose 118.6 mg/g. The result provided the potential of roots of L. lucidus Turcz. as new raffinose family oligosaccharides sources for functional food. Moreover, since the present systematic strategy is efficient, sensitive and robust, separation, purification and quantification of oligosaccharides by hydrophilic interaction liquid chromatography seems to be possible.

Lycopus lucidus Turcz. ex Benth. var. hirtus Regel (Lamiaceae) in Italy: a new naturalized alien species for the European flora

Lycopus lucidus Turcz. ex Benth. var. hirtus Regel (Lamiaceae), a neophyte native to Asia, is recorded for the first time in Europe. A naturalized population was discovered in the Milan railway area (Italy). Diagnostic characters and an updated identification key for the genus Lycopus in Europe are presented. Furthermore, the invasive status and the possible means of introduction are discussed.

Ethanol Extract ofLycopus lucidusElicits Positive Inotropic Effect Via Activation of Ca2+Entry and Ca2+Release in Beating Rabbit Atria

Lycopus lucidus Turcz has been widely used as a traditional Oriental medicine (TOM) in Korea and China and prescribed for the enhancement of heart function. However, the precise effects have yet to be defined. The purpose of the present study was, therefore, to address whether the ethanol extract of Lycopus lucidus Turcz (ELT) has a positive inotropic effect. ELT-induced changes in atrial mechanical dynamics (pulse pressure, dp/dt, and stroke volume), and cAMP efflux were measured in perfused beating rabbit atria. Three active components, rosmarinic acid, betulinic acid, and oleanolic acid were identified in ELT by high performance liquid chromatography analysis. ELT increased atrial dynamics in a concentration-dependent manner without changes in atrial cAMP levels and cAMP efflux. The ELT-induced positive inotropic effect was blocked by inhibition of the L-type Ca(2+) channels and sarcoplasmic reticulum (SR). Inhibitors of β-adrenoceptors had no effect on the ELT-induced positive inotropic effect. The results suggest that ELT exerts a positive inotropic effect via activation of Ca(2+) entry through L-type Ca(2+) channel and Ca(2+) release from the SR in beating rabbit atria.

택란의 염색성 및 항균성

Abstract In this research, the dyeing properties of Lycopus lucidus Turcz on cotton and silk fabrics were evalu-ated to establish a scientific database of natural dyes. The optimum dyeing conditions were identified withK/S values depending on the dyeing concentration, temperature, time, and pH. For the optimum con-ditions, the color changes for different synthetic mordants were observed as L, a*, b* and H, V, C. Thecolor fastness after dyeing and mordanting was also examined. Additionally, the antibiosis of Lycopuslucidus Turcz was examined. The results were as follows: For the cotton, the optimized dyeing conditionswere 300% (o.w.f.), 50 o C, 40 min., and pH 5. For the silk fabric, the conditions were 450% (o.w.f.), 70 o C,60 min., and pH 4. The K/S value of dyed cotton improved about 1.03-2.78 folds after mordanting. Thefabric color was yellow in the absence of mordanting, while it was reddish yellow after post-mordantingwith FFC. Although the color changed with the mordanting method and mordant, the overall hue wasyellow. Thus, Lycopus lucidus Turcz appears to have the potential for use as a natural dyeing material fora yellow hue. The K/S value of the dyed silk fabric improved about 1.05-1.40 folds after mordanting. Thefabric showed a reddish yellow color when pre-mordanted with Al and Cu. The washing and dry-cleaningfastness of the cotton and silk fabrics were at the 4-5 level. The light fastness of the cotton improved aftermordanting; however, pre-mordanting was more effective than post-mordanting or the absence of mor-danting. The dyed cotton, silk fabrics showed excellent antibiosis.Key words: Lycopus lucidus Turcz, Dyeability, Antimicrobial activity, Mordant, Color fastness; 택란, 염색성, 항균성, 매염, 염색견뢰도

Anticancer, antioxidant and antimicrobial activities of the essential oil of Lycopus lucidus Turcz. var. hirtus Regel

This study was designed to examine the anticancer, antioxidant and antimicrobial activities of the essential oil from Lycopus lucidus Turcz. var. hirtus Regel. The essential oil treatment to six human cancer cell lines resulted in a dose-dependent inhibition of cell growth. The cytotoxicity of the essential oil on liver carcinoma and breast cancer cell lines was significantly stronger than on other cell lines. The essential oil can induce apoptosis of the liver carcinoma cell line Bel-7402 and decrease the intracellular GSH level. The antioxidant effect of the essential oil was evaluated by using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and hydroxyl radical (OH) scavenging assays. The essential oil exhibited moderate antioxidant activity. The antimicrobial activity of the essential oil was evaluated against eight microorganisms using the disc diffusion and broth microdilution methods. The essential oil also showed moderate antimicrobial activity. These suggest that the essential oil could hold a good potential for use in the pharmaceutical industry.

Isolation, Characterization, and Immunological Effects of α-Galacto-oligosaccharides from a New Source, the Herb Lycopus lucidus Turcz.

This study was designed to isolate and characterize a mixture of alpha-galacto-oligosaccharides (GOS) from a new source, the roots of Lycopus lucidus Turcz. (RL), a traditional dietary treatment. In this study, the chemical components and immunological function of RL-GOS were investigated. HPLC analysis showed that the purified RL-GOS was a typical raffinose family oligosaccharide (RFO) with a high stachyose content of 51.8% (w/w), followed by 26.5% raffinose and 10.1% verbascose. Further functional evaluation showed that RL-GOS could elicit a significant increase (p < 0.05 vs control) in humoral immunity, as measured by plaque-forming cell (PFC) generation and serum hemolysin level in response to sheep red blood cells (SRBC) at all three tested doses of RL-GOS (0.75, 1.5, and 3.0 g/kg of BW) in mice. In addition, the cellular immune activity of RL-GOS was also demonstrated by enhancing in vivo delayed-type hypersensitivity (DTH) reaction to SRBC and spleenocyte proliferation response to concanavalin A (p < 0.05, compared with control group). Nevertheless, there were no significant differences in weight gain, lymphoid organ indices, and phagocytosis capacity following RL-GOS treatment. This study provides evidence for the discovery of a new GOS source (20% w/w GOS in fresh roots of L. lucidus Turcz.) and its potential application as an immune stimulant in functional foods.

Effect of betulinic acid on high glucose‐induced vascular smooth muscle cells proliferation

Betulinic acid (BA) is a pentacyclic triterpene found in Lycopus lucidus Turcz. Hyperglycemia has been shown to stimulate vascular smooth muscle cells proliferation, a paramount feature in atherosclerosis. The aim of this study was to determine the effects of BA on high glucose‐induced human aortic smooth muscle cell (HASMC) proliferation. In this study, [3H]‐thymidine incorporation under 25 mM glucose (HG) was significantly accelerated compared with 5.5 mM glucose and this increase was inhibited by BA. Invasion and migration assay also suggested an anti‐proliferative effect of BA. BA elevated the protein content of p21waf1/cip1, p27kip1, decreased cyclin D1/E and CDK2/4, suggesting that BA inhibited HASMC proliferation by regulating the cell progression from G1 to S phase. We showed that BA significantly inhibited HG‐induced matrix metalloproteinase (MMP)‐2/‐9 activity in a dose‐dependent manner. BA inhibited the HG‐induced hydrogen peroxide production and DCF‐sensitive intracellular ROS formation. Furthermore, BA suppressed the translocation and promoter transcriptional activity of NF‐κB under HG condition. BA also decreased HG‐induced phosphorylation of IκB‐α. In conclusion, the present data suggested that BA regulated cell cycle regulatory proteins, and significantly reduced HASMC proliferation, via inhibition of ROS and NF‐κB activation in hyperglycemia conditions.