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-1ß 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.

Anti-tumor effect of Yanggan Huayu granule by inducing AKT-mediated apoptosis in hepatocellular carcinoma.

ETHNOPHARMACOLOGICAL RELEVANCE: Yanggan Huayu granule (YGHY) is a formula of traditional Chinese medicine that has been widely used to treat patients with liver cancer. But its working mechanism is still poorly understood. AIM OF THE STUDY: To investigate the anti-tumor effect of YGHY and its working mechanisms in hepatocellular carcinoma (HCC). MATERIALS AND METHODS: H22 mouse xenograft model was used to detect the effect of YGHY on hepatocellular carcinoma (HCC). MTT and CCK8 assays were performed to assess the effect of YGHY on HCC cell growth. Transwell assay was performed to detect the invasion and migration activities of HCC cells. Effect of YGHY drug-contained serum on apoptosis was detected by flow cytometry. Western blot was performed to detect the protein expressions. RESULTS: Results showed that YGHY inhibited tumor volume and weight, induced the apoptosis of HepG2 and SMMC-7721 cells and increased the protein expressions of Cleaved-Caspase3 and Cleaved-PARP. Furthermore, YGHY significantly down-regulated the protein expression of p-AKT. SC79, as an activator of AKT signaling, was able to increase the expression of p-AKT, and regulate the protein expressions of Cleaved-Caspase3, Cleaved-PARP, BCL-2 and BAX. YGHY drug-contained serum negated the protein expression change provided by SC79. CONCLUSIONS: Taken together, this data indicates that YGHY could inhibit HCC growth by inducing apoptosis, operating through AKT signaling.

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

AIMS AND OBJECTIVE: 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. MATERIALS AND METHODS: 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. RESULTS: 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. CONCLUSION: 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 ameliorates DNCB­induced 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 anti­inflammatory, antioxidant, anti­vascular inflammation and wound­healing 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 2,4­dinitrochlorobenzene (DNCB)­induced dermatitis and to verify the inhibitory effect of LLT on the expression of chemokines and pro­inflammatory cytokines in the HaCaT immortalized keratinocyte cell line. In addition, the anti­inflammatory function of LLT in RAW264.7 mouse macrophages was investigated. In the DNCB­induced 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 IL­6 in serum. In addition, LLT inhibited the phosphorylation of ERK and JNK, as well as NF­κB in skin tissue. In the HaCaT cell model induced by TNF­α/IFN­Î³, LLT inhibited the expression of thymus and activation­regulated chemokine, granulocyte­macrophage colony­stimulating factor, monocyte chemoattractant protein­1, TNF­α and IL­1ß, whilst inhibiting the phosphorylation of NF­κB. In addition, in the lipopolysaccharide­induced RAW 264.7 cell inflammation model, LLT inhibited the expression of TNF­α and IFN­Î³, the nuclear translocation of NF­κB and the phosphorylation of ERK and JNK. These results suggested that LLT may be a promising candidate for the treatment of inflammatory dermatitis.

Antimicrobial and Antioxidant Properties of Four Lycopus Taxa and an Interaction Study of Their Major Compounds.

The compositions of leaf infusions of three genotypes of Lycopus europaeus L. with origins in central Europe, namely L. europaeus A (LeuA), L. europaeus B (LeuB), and L. europaeus C (LeuC), and one genotype of L. exaltatus (Lex), were examined by LC-MS-DAD (Liquid Chromatography Mass Spectrometry and Diode Array Detection) analysis. This revealed the presence of thirteen compounds belonging to the groups of phenolic acids and flavonoids, with a predominance of rosmarinic acid (RA) and luteolin-7-O-glucuronide (LGlr). The antimicrobial activity of leaf infusions was tested on the collection strains of Gram-positive and Gram-negative bacteria, and on the clinical Staphylococcus aureus strains. We detected higher activity against Gram-positive bacteria, of which the most susceptible strains were those of Staphylococcus aureus, including methicillin-resistant and poly-resistant strains. Furthermore, we examined the antioxidant activity of leaf infusions using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) methods, and on NIH/3T3 cell lines using dichlorodihydrofluorescein diacetate (DCFH-DA). We also studied the mutual interactions between selected infusions, namely RA and/or LGlr. In the mixtures of leaf infusion and RA or LGlr, we observed slight synergism and a high dose reduction index in most cases. This leads to the beneficial dose reduction at a given antioxidant effect level in mixtures compared to the doses of the parts used alone. Therefore, our study draws attention to further applications of the Lycopus leaves as a valuable alternative source of natural antioxidants and as a promising topical antibacterial agent for medicinal use.

Inhibitory activities of phenylpropanoids from Lycopus lucidus on amyloid aggregation related to Alzheimer's disease and type 2 diabetes.

The number of patients with Alzheimer's disease (AD) and type 2 diabetes (T2D) is increasing rapidly, and thus more research has been focused on the relationship between these two age-related chronic diseases. According to the amyloid hypothesis, prevention of the aggregation of amyloid ß (Aß) and human islet amyloid polypeptide (hIAPP) is a promising strategy for AD and T2D. In this study, thioflavin-T assay and transmission electron microscopy were performed to evaluate the inhibitory effect of three phenylpropanoids isolated from Lycopus lucidus-schizotenuin A and lycopic acids A and B-on both Aß and hIAPP fibrillization. All tested compounds exhibited similarly strong inhibitory activity toward amyloid aggregation. These results suggested that catechol moieties play important roles in the inhibition of amyloid plaque formation.

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.

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.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is closely related to metabolic diseases such as obesity and insulin resistance. PURPOSE: We studied whether an ethanol extract of Lycopus lucidus Turcz. ex Benth (LLE) exhibited effects on lipid metabolism in NAFLD. STUDY DESIGN: An 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). METHODS: HepG2 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. RESULTS: LLE significantly decreased the intracellular lipid accumulation in FFA-treated HepG2 cells. LLE not only remarkably decreased the expression of lipogenesis genes but also increased ß-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. CONCLUSION: These results suggest that LLE may exert protective effects against NAFLD-related obesity and metabolic disease.

Identification of the NADPH Oxidase 4 Inhibiting Principle of Lycopus europaeus.

NADPH oxidase 4 (Nox4) has recently been implicated as driving force in cellular senescence. Thus, there is growing interest to develop Nox4 inhibitors, which might be valuable agents for cosmeceutical applications. Alpine plants represent a valuable source for the identification of novel bioactive natural products with anti-ageing effects, especially substances that protect plants against UV radiation, which is also known to contribute to the ageing of human skin. Therefore, the aim of this study was to identify novel Nox4 inhibitors from alpine plants. Within an initial screening of extracts of alpine plants on their ability to inhibit Nox4 activity in HEK cells, the methanolic extract of the subaerial parts of Lycopus europaeus showed a strong inhibition of Nox4 (81% chemiluminescence quenching) and a simultaneously high cell viability (91% vitality). Rosmarinic acid was isolated and identified as the major compound in this bioactive extract. It showed a dose dependent inhibitory activity on Nox4 with an IC50 of 1 µM. Moreover, it also showed a significant inhibitory activity on Nox2 in the low micromolar range, whereas no inhibition of Nox5 was detected. Further investigations confirmed that the observed effects of rosmarinic acid on Nox2 and Nox4 are real inhibitory activities, and not due to ROS scavenging effects. Therefore, L. europaeus, which we demonstrated to be a good source of rosmarinic acid, has great potential for usage in cosmeceutical products with anti-ageing activity.

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.

Pancreatic lipase and α-amylase inhibitory activities of plants used in Traditional Chinese Medicine (TCM).

To find new, plant based drugs for the treatment of obesity and/or diabetes mellitus type 2 through the inhibition of essential digestive enzymes, in vitro tests were carried out on selected plants or fungi with weight-reducing, blood glucose-reducing or related potential, used in Traditional Chinese Medicine (TCM). Aqueous and methanolic extracts of 32 Chinese herbal medicines were assayed for their in vitro inhibitory activity against pancreatic lipase (PL) and α-amylase (PA). PL activity was measured by using an enzymatic in vitro assay based on the hydrolysis kinetics of an oleate ester of 4-methylumbelliferone. For the determination of α-amylase activity an enzyme assay based on the hydrolytic cleavage of a modified starch derivative was used. Our findings have shown that the methanolic extract of Lycopus lucidus Turcz. var. hirtus Regel (Lamiaceae) was a very effective PL inhibitor (IC50: 88.3±4.1 µg/mL). A high anti-amylase activity showed the methanolic extract of Trichosanthes kirilowii Maxim. (Curcurbitaceae, IC50: 248.8±67.3 µg/mL). This work provides a priority list of interesting plants for further study with respect to the treatment of obesity and associated metabolic diseases.

Ditch network sustains functional connectivity and influences patterns of gene flow in an intensive agricultural landscape.

In intensive agricultural landscapes, plant species previously relying on semi-natural habitats may persist as metapopulations within landscape linear elements. Maintenance of populations' connectivity through pollen and seed dispersal is a key factor in species persistence in the face of substantial habitat loss. The goals of this study were to investigate the potential corridor role of ditches and to identify the landscape components that significantly impact patterns of gene flow among remnant populations. Using microsatellite loci, we explored the spatial genetic structure of two hydrochorous wetland plants exhibiting contrasting local abundance and different habitat requirements: the rare and regionally protected Oenanthe aquatica and the more commonly distributed Lycopus europaeus, in an 83 km(2) agricultural lowland located in northern France. Both species exhibited a significant spatial genetic structure, along with substantial levels of genetic differentiation, especially for L. europaeus, which also expressed high levels of inbreeding. Isolation-by-distance analysis revealed enhanced gene flow along ditches, indicating their key role in effective seed and pollen dispersal. Our data also suggested that the configuration of the ditch network and the landscape elements significantly affected population genetic structure, with (i) species-specific scale effects on the genetic neighborhood and (ii) detrimental impact of human ditch management on genetic diversity, especially for O. aquatica. Altogether, these findings highlighted the key role of ditches in the maintenance of plant biodiversity in intensive agricultural landscapes with few remnant wetland habitats.

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.

Variation in nutritional compositions, antioxidant activity and microstructure of Lycopus lucidus Turcz. root at different harvest times.

The objective of this study was to investigate the variation in nutritional compositions, antioxidant activity and microstructure of Lycopus lucidus Turcz. root at different harvest times. L. lucidus Turcz. roots, harvested from two sites (S1 and S2) at three different times (T1: 19-11-2013, T2: 22-12-2013 and T3: 27-01-2014), were analyzed for nutritional compositions, antioxidant activity by DPPH, FRAP and TEAC assays and microstructure. The results revealed that the protein content in L. lucidus Turcz. root first decreased and then increased to a maximum at T3. The reducing sugar content had no significant differences among the three harvest dates studied. The starch content decreased drastically along with an increase of crude fat content with the harvest time delayed. The major amino acids in L. lucidus Turcz. root were aspartic acid and glutamate and the highest total amino acid content was found for the root harvested at T3. The most common element in L. lucidus Turcz. root was detected to be potassium followed by calcium, iron, magnesium, copper and manganese, and their changes were discrepant in the period of harvest. The FP and SGP possessed the highest and lowest phenolic content, respectively. The change of SEP was significantly correlated to the SGP at different harvest times. The highest TPC was found for the root harvested at T3 and the most abundant phenolic acid was chlorogenic acid. The highest and lowest DPPH radical scavenging capacity was observed for the SGP and FP, respectively. The highest and lowest FRAP and TEAC were observed for the FP and SGP, respectively. The results of correlation analysis indicated that there was significant correlation between phenolic content and FRAP and TEAC, and different antioxidant assays. The microstructure of L. lucidus Turcz. root also varied greatly with the harvest times.

Lycopus europaeus: phenolic fingerprint, antioxidant activity and antimicrobial effect on clinical Staphylococcus aureus strains.

Lycopus europaeus L. leaves water extract (LEL) was subjected to phytochemical analysis, and evaluated for its antibacterial and antioxidant effects. Antibacterial activity testing was performed on Staphylococcus aureus clinical strains from catheter-related and skin infections by broth microdilution test. LEL showed bactericidal activity at concentrations from 2500 to 5000 µg/mL against all, including methicillin resistant and polyresistant nosocomial, strains. Antioxidant activity was examined using DPPH and ABTS (11.3 and 9.8 µg/mL, respectively) and by ferric reducing ability of the plasma method (891 µmol AAE/g dry extract). Phytochemical analysis of LEL was performed by LC-DAD-MS/MS. Ten phenolic compounds were identified; two minor compounds (glucopyranosyl rosmarinic acid and sagerinig acid) have not been described in Lycopus yet. The major compounds, considered to be responsible for biological activities detected in the study, were determined as rosmarinic acid (76 mg/g) and luteolin-7-O-glucuronide (23 mg/g). L. europaeus arises from our study as a promising source of antibacterial agent for topical usage.

Stachyose-enriched α-galacto-oligosaccharides regulate gut microbiota and relieve constipation in mice.

This study probed the effects of Deshipu stachyose granules (DSG), a novel oligosaccharide preparation (55.3% stachyose, 25.8% raffinose, and 9.7% verbascose), on gut microbiota and constipation in mice. Mice were administered intragastrically without or with DSG (0.42, 0.83, and 2.49 g/kg bw), and feces were collected after 14 days of treatment and subjected to classical microbiological assays. Selective index (SI) and prebiotic index (PI) were incorporated to evaluate the prebiotic effect. DSG at 0.83 g/kg bw scored the highest SI and PI scores, thus supporting a strong prebiotic role. In addition, the impact of DSG (0.42, 0.83, and 1.68 g/kg bw) on defecation function of constipated mice was determined. Ink propulsion rate in the small intestine was significantly improved by DSG treatment. DSG supplementation also distinctly increased the weight and number of black feces within 5 h and evidently shortened the defecating time of first black feces, as compared with the constipation control mice. All of these findings indicate that DSG may promote the growth of beneficial intestinal bacteria and inhibit pathogenic bacteria and also facilitate intestinal peristalsis and fecal excretion, thereby enhancing intestinal health and relieving constipation.

The aqueous extract of Lycopus lucidus Turcz ameliorates streptozotocin-induced diabetic renal damage via inhibiting TGF-ß1 signaling pathway.

PURPOSE: Renal fibrosis characterized by accumulation of extracellular matrix protein results in chronic renal diseases including diabetic nephropathy. Transforming growth factor ß1 (TGF-ß1) signaling pathway plays a key role in mediating renal fibrosis. Hence, agents that antagonize TGF-ß signaling could be candidate for kidney disease therapy. METHODS: We established renal fibrosis model both in vitro with fibroblast cells treated with rhTGF-ß1 and streptozocin(STZ)-induced diabetic nephropathy rats model in vivo and evaluated the effect of the aqueous extract of Lycopus lucidus Turcz, the blood-circulation-promoting Chinese herb, on diabetic nephropathy and investigated the mechanism of action. RESULTS: We found that Lycopus suppressed rhTGF-ß1-induced Smad2 and ERK1/2 activation, down-regulated the expression of TGF-ßRI, TGF-ßRII, Smad4 and Smad7 in SV40 MES13 cells without inhibiting cell viability. In vivo, lycopus inhibited Smad2 phosphorylation, reduced mRNA level of TGF-ß1, ameliorated expansion of the mesangial area in glomerular tissue and reduced the levels of Scr and BUN of serum and total-SOD (superoxide dismutase) activity in STZ-induced diabetic rats. CONCLUSION: Lycopus is a novel inhibitor of renal fibrosis by blocking TGF-ß signaling pathway and possess a protective effect on renal damage of STZ-induced diabetic nephropathy in rats.

Ethanol extract of Lycopus lucidus elicits 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 ß-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.

High dilutions of homeopathic remedies induce relaxation of rat aorta precontracted with noradrenalin

BACKGROUND Homeopathic potencies have been reported to produce alteration of contraction in isolated rat ileum in an organ bath. Potentized homeopathic drugs like Lycopus V and Aurum met are used for the treatment of hypertension. AIM The purpose of this study is to see whether Lycopus V 30 CH and Aurum met 30 CH could produce relaxation of isolated rat aorta in the organ bath. METHODS The aorta of rats were dissected out, placed in Krebs-Henseleit solution, cleared of connective tissue and endothelium and cut into 2-2.5 mm long rings. The rings were fixed in organ baths with the upper end connected by a string to an isometric transducer which was finally attached through a data acquisitation equipment to a computer. Aurum met 30 CH Lycopus V 30 CH, and their medium 90% ethanol were added separately to the bathing fluid containing the aorta rings which were precontracted with noradrenalin (NA). RESULTS Both the drugs produced significant relaxation of the aorta (p<0.001) precontracted with NA (10-7 M). The control did not show any marked effect on the NA induced contraction of aorta. CONCLUSION A potentised drug is thought to be specifically structured water which can transform the water structure in the bathing fluid and the isolated tissue immersed in the fluid into its own form. This in turn produces the observed relaxation. Both Lycopus V 30 CH and Aurum met 30 CH are effective in reducing NA induced contraction of rat aorta. Drugs can directly act on the isolated rat aorta without any system of influence. KEY WORDS High dilution drug, isolated aorta, hypertension, noradrenalin. (AU)

Introdução e objetivo: Segundo alguns estudos, diluições homeopáticas alteram a contratilidade de íleo isolado de rato, em banho de órgãos. Os medicamentos homeopáticos como Aurum metallicum Lycopus virginicus são usados no tratamento da hipertensão. O objetivo deste estudo foi determinar se Lycps 30 cH e Aur 30 cH podem induzir o relaxamento da aorta isolada de ratos, em banho de órgãos. Métodos: A aorta foi dissecada e, livre de tecido conjuntivo e endotelio, foi colocada em solução de Krebs-Henseleit, sendo seccionada em anéis de 2 a 2,5 cm de comprimento. Os anéis foram fixados em banho de órgãos, a extremidade superior ligada por um cabo a um transdutor isométrico, por sua vez ligado a dispositivos de registro de dados em um computador. Aur 30 cH, Lycps 30 cH e etanol 90%, foram adicionados separadamente ao líquido do banho contendo os anéis de aorta previamente contraídas pela noradrenalina (NA) ou norpinefrina (NE). Resultados: Os dois medicamentos testados induziram relaxamento significativo (p <0,001), do preparada de aorta contraída por NA (10-7 M), enquanto que a solução de controle não produziu efeitos significativos. Conclusão: Ambas as drogas foram usadas em diluições homeopáticas demasiado elevadas para admitir a presença de moléculas da substância original. Assim, o mecanismo farmacológico tradicional envolvendo as moléculas da droga e receptores no músculo liso da aorta deve ser descartado. Embora reconhecidamente diferente, o mecanismo envolvido permanece desconhecido. Lycps 30 e Aur 30 CH mostraram-se eficazes na redução da contratilidade da aorta de ratos, induzida por NA. As drogas podem agir diretamente na aorta isolada de ratos sem influência sistêmica. (AU)