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.

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.

Anti-tumor Effect and Mechanism of Lycopus lucidus Polysaccharide on Non-small Cell Lung Cancer A549 Cells / 中国实验方剂学杂志

ObjectiveTo study the anti-tumor activity and mechanism of Lycopus lucidus polysaccharide （LLP） in vitro. MethodCell counting kit-8 （CCK-8） assay was used to detect the inhibitory effect of LLP （0， 5， 10， 15， 20 g·L-1） on the proliferation of A549 cells at different time points （24，48，72 h）. The migration and invasion abilities of A549 cells were detected by wound healing assay and transwell assay after LLP （10， 20 g·L-1） treatment for 24，48 h. Propidium iodide （PI） single staining was applied to determine the effect of LLP of different concentrations （10，20 g·L-1） on the cell cycle of A549. The apoptosis of A549 cells induced by LLP （10， 20 g·L-1） was detected by Annexin V-FITC/PI kit. Real-time fluorescence-based quantitative polymerase chain reaction （Real-time PCR） was adopted to measure effect of LLP （10， 20 g·L-1） on gene expression of cysteine aspartate protease-3 （Caspase-3），cysteine aspartate protease-8 （Caspase-8），cysteine aspartate protease-9 （Caspase-9），cyclin-dependent kinase-1 （CDK-1）， and Cyclin B1 in A549 cells. Western blot was used to detect the effect of LLP on protein expression of Caspase-3，Caspase-8，Caspase-9，B-cell lymphoma 2 （Bcl-2）-associated X protein （Bax），CDK-1，cyclin-dependent kinase-4 （CDK-4），cyclin-dependent kinase-6 （CDK-6），Cyclin B1，and Cyclin D1 in A549 cells. ResultCompared with the blank group， the LLP group showed decreased proliferation， migration， and invasion of A549 cells （P<0.05， P<0.01）， increased proportion of G0/G1 phase （P<0.05）， enhanced apoptosis rate （P<0.05， P<0.01）， elevated mRNA expression of Caspase-3，Caspase-8，and Caspase-9 （P<0.05，P<0.01）， reduced mRNA expression of CDK-1 and Cyclin B1 （P<0.05，P<0.01）， up-regulated protein expression of Caspase-3，Caspase-8，Caspase-9， and Bax （P<0.05， P<0.01）， and down-regulated protein expression of Bcl-2， CDK-1， CDK-4， CDK-6， Cyclin B1， and Cyclin D1 （P<0.05， P<0.01）. ConclusionLLP can inhibit the proliferation of A549 cells， block the cell cycle in the G0/G1 phase （also G2/M phase）， and induce cell apoptosis via the mitochondrial apoptosis pathway and death receptor pathway.

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.

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

PURPOSE: Lycopus 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 METHODS: Primary 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-1ß, 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. RESULTS: LLT 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-1ß/IL-18. We demonstrated that LLT enhances axonal elongation and synaptic connectivity upon H2O2-induced neuronal injury in rat primary cortical neurons. CONCLUSION: It 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.

In Vitro and In Silico Studies of Soluble Epoxide Hydrolase Inhibitors from the Roots of Lycopus lucidus.

Soluble epoxide hydrolase (sEH) is an enzyme that is considered a potential therapeutic target in human cardiovascular disease. Triterpenes (1-4) and phenylpropanoids (5-10) were isolated from Lycopus lucidus to obtain sEH inhibitors through various chromatographic purificationtechniques. The isolated compounds were evaluated for their inhibitory activity against sEH, and methyl rosmarinate (7), martynoside (8), dimethyl lithospermate (9) and 9″ methyl lithospermate (10) showed remarkable inhibitory activity, with the IC50 values ranging from 10.6 ± 3.2 to 35.7 ± 2.1 µM. Kinetic analysis of these compounds revealed that 7, 9 and 10 were competitive inhibitors bound to the active site, and 8 was the preferred mixed type inhibitor for allosteric sites. Additionally, molecular modeling has identified interacting catalytic residues and bindings between sEH and inhibitors. The results suggest that these compounds are potential candidates that can be used for further development in the prevention and treatment for cardiovascular risk.

The complete chloroplast genomes of Lycopus lucidus and Agastache rugosa, two herbal species in tribe Mentheae of Lamiaceae family.

Lamiaceae is one of the largest families in the kingdom Plantae, including lots of traditional Chinese herbs. Lycopus lucidus and Agastache rugosa are two Lamiaceae species, which are most frequently used in Chinese traditional medicine. In the current study, the complete chloroplast genome sequences of two species were assembled. Their circular DNA lengths were 152,096 and 151,922 bp respectively. Both genomes were made up of a large single-copy region, a small single-copy region, and a pair of inverted repeat regions. Each genome totally encoded 133 genes, containing 88 protein-coding genes, 37 tRNA genes, and eight rRNA genes. Phylogenetic analysis indicated that both species belong to the Mentheae tribe of the Lamiaceae family.

Simultaneous Determination of 7 Phenolic Acids in Lycopus lucidus var. hirtus Rhizome by HPLC-PDA / 中国实验方剂学杂志

Objective:A high performance liquid chromatography-photo-diode array（HPLC-PDA） method for the simultaneous determination of the 7 phenolic acids including danshensu，protocatechuic acid，protocatechuic aldehyde，chlorogenic acid，caffeic acid，ferulic acid and rosemary acid in Lycopus lucidus var.hirtus rhizome，analyzing and evaluating the phenolic acids in L.lucidus var.hirtus rhizome collected from different habitats，is reported here. Method:The sample was extracted by ultrasonic with 80% methanol solution，7 kinds of phenolic acids were separated on a CAPCELL PAK C18 column （4.6 mm×250 mm，5 μm） with a mobile phase consisting of methanol-0.02% formic acid aqueous （pH 3.10）by gradient elution，The detection wavelength was at 279，324 nm， the column temperature was 30 ℃ with 20 μL injection volume and the flow rate was 1.0 mL·min-1. Result:The 7 Phenolic acids had a good linear relationship （r≥0.999 9） within their respective mass concentration ranges，the average recovery was 96.49%-103.45% and the RSD was 0.5%-2.8%，the limit of determination was 0.008-0.046 mg·L-1 and the limit of quantification was 0.027-0.154 mg·L-1.The 7 kinds of phenolic acids were all detected in L.lucidus var.hirtus rhizome and the total amount was between 5 811.01 and 11 747.23 µg·g-1 ， the average amount was 7 421.05 µg·g-1.The content of 7 phenolic acids was different and the rosemary acid was the highest in all the samples with an average of 7 111.19 µg·g-1 the ratio to the total phenolic acids was 95.8%.The results of principal component analysis and cluster analysis showed that the quality of L.lucidus var.hirtus rhizome from Heze city in Shandong province was better，followed by Wanzhou district in Chongqing. Conclusion:The method was simple，sensitive，accurate，practical and reliable，and is suitable for the content determination of phenolic acid in L. lucidus var. hirtus rhizome.It is expected to provide a reference for the improvement of quality standard and a new idea for the development and utilization of L.lucidus var.hirtus rhizome.

The aqueous extract of Lycopus lucidus Turcz exerts protective effects on podocytes injury of diabetic nephropathy via inhibiting TGF-ß1 signal pathway.

Diabetic nephropathy (DN) is known as a major microvascular complication leading cause of end-stage renal disease, it generally followed by the process of podocyte fragmentation and detachment. Transforming growth factor ß1 (TGF-ß1) signaling pathway plays a pivotal role in the initiation and progression of DN. In present study, we aim to investigate the effect of lycopus extracts on podocytes injury and TGF-ß signaling. In present study, lycopus extracts treatment abolished the gain in blood glucose and body weight in a dose dependent manner and possessed protective effect on the renal damage, which was indicated by the decreased concentration of Scr, BUN and urine creatinine of serum. Histopathological examination also demonstrated lycopus extracts exert protective effect on renal damage. Western blotting and immunohistochemical results revealed lycopus extracts treatment upregulated the expression of nephrin and down-regulated the expression levels of TGF-ß1 and Smad4. Moreover, lycopus extracts treatment suppressed TGF-ß1-induced phosphorylation of Smad2/3, ERK1/2 and p38 both in vivo and vitro. In conclusion, lycopus extracts is a novel agent that ameliorate podocytes injury by inhibiting TGF-ß signaling pathway and possess potential therapeutic effect on renal damage of DN rats.

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.

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.

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.

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.

Protective effect of Lycopus lucidus Turcz ethanol extract on kidney in STZ induced diabetic mice / 中国生化药物杂志

Objective To study the protective effect and mechanism of Lycopus lucidus Turcz ethanol extract on STZ induced diabetic mice kidney.Methods The diabetic mice model were induced by single intraperitoneal injection of 0.12 mg/g STZ.After 60h, the mice successful modeling were divided into model control group,Lycopus lucidus Turcz ethanol extract high-dose group [5.0g/(kg? d)]and low-dose group [1.25/(kg? d)], 10 mice in each group.Another 10 normal mice were used as normal control group.The high-and low-dose group were intragastric administrated corresponding dose Lycopus lucidus Turcz ethanol extract, normal control group and model control group were given the same volume of sterile distilled water.After 5 weeks, the mice kidney structure was observed by periodic acid-Schiff ( PAS ) , advanced glycosylation end products ( AGEs ) and transforming growth factorβ1 (TGF-β1) in kidney tissue were detected by ELISA.The monocyte chemotactic protein-1 (MCP-1) mRNA expression were detected by RT-PCR and MCP-1 protein expression by Western blot in mice kidney.Results PAS staining results showed, compared with model control group,renal structural changes in high-dose group was significantly increased.ELISA results showed AGEs and TGF-β1 content in kidney tissue of model control group were higher than normal control group (P<0.01), the above indexes of high-dose group were lower than model control group (P<0.05) .RT-PCR results showed MCP-1 mRNA expression of model control group was higher than normal control group (P<0.01), MCP-1 mRNA expression of low-and high-dose group model group were lower than model control group (P<0.01), and MCP-1 mRNA expression of high-dose group was lower than low-dose group (P<0.05).Western blot results showed MCP-1 protein expression of model group was higher than normal control group (P<0.01), but there were no significant differences of MCP-1 protein expression between low-or high-dose group model group and model control group. Conclusion Lycopus lucidus Turcz ethanol extract can protect the STZ-induced diabetic mice kidney, and it might be the reason of inhibiting the expression of AGEs-MCP-1-TGF-β1.

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.

Identification of compounds in aqueous extract from aerial parts of Lycopus lucidus by HPLC-ESI-TOF-MS/MS / 中草药

Objective: To identify the main active compounds in the aqueous extract from the aerial parts of Lycopus lucidus by high performance liquid chromatography (HPLC)-electrospray (ESI)-time of flight tandem mass spectrometry (TOF-MS/MS). Methods: The aqueous extract from the aerial parts of L. lucidus was prepared using ultrasonic method; Chromatographic separation of the main active compounds was performed on a TC-C18 (250 mm × 4.6 mm, 5 μm) reverse phase column through gradient elution; All the compounds eluted from the column were detected under both positive and negative ionization modes. Each chromatographic peak was analyzed by quadrupole tandem mass spectrometry coupled with TOF. Results: Twenty two compounds were identified through the analysis of tandem mass spectrum and the information from reference substances, including amino acids, phenolic acids, terpenoids, flavonoids and flavonoid glycosides, sterols, and fatty acid. Conclusion: HPLC-ESI-TOF-MS/MS is capable of analyzing the main compounds in the aerial parts of L. lucidus using retention time, ultraviolet spectrum, current molecular weight, formula, and fragmenting information of daughter ions. It will probably become a reliable alternative for the rapid analyzing substantial foundation of Chinese materia medica.

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.

Effects of active fractions L.F04 from ground part of Lycopus lucidus var.hirtus on platelet aggregation and thrombosis formation / 中草药

Object To study the effects of active fraction L.F04 from the ground part of Lycopus lucidus Turcz. var. hirtus Reg. on platelet aggregation and thrombosis formation and investigate its mechanisms of promoting blood circulation and removing blood stasis. Methods The effects of L.F04 on platelet aggregation induced by ADP in vivo, thrombosis of artery vein side road and thrombus formed in rotary loop in vitro were examined, the rat model of blood stasis made by injecting high molecular weight dextran (HMWD) was used. Results L.F04 0.408 and 0.204 g/kg evidently inhibited the ADP induced increase of platelet maximum aggregation rate in vivo in HMWD model, with a concentration dependent manner. Compared with the control group, the thrombus weight in rat model of blood stasis was increased significantly and the length of thrombus was shown an increasing trendency. L.F04 0.408 and 0.204 g/kg both shown the anti thrombosis effect. L.F04 0.408 g/kg showed better effects of lessening the thrombus dry weight and wet weight. Both L.F04 0.408 and 0.204 g/kg could inhibit the thrombosis of artery vein side road, the inhibition rates are separately 27.41% and 27.14%. Conclusion L. lucidus var. hirtus F04 could significantly inhibit platelet aggregation and thrombosis formation.