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Abstract TCMSP platform of systematic pharmacology of traditional Chinese medicine This study aimed to investigate the molecular mechanism of Fructus Ligustri Lucidi (NZZ, Chinese abbreviation) against osteoporosis (OP) by means of network pharmacology.ChemDraw Professional 15.1 software and Molinspiration Smiles database were used to draw the chemical formulas of the components. The active ingredients and related target proteins of NZZ were searched in platform of systematic pharmacology of traditional Chinese medicine database, Drugbank, Therapeutic Target Database, SymMap and other databases. Gene Ontology(GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were carried out on the selected target through Enrichr and KEGG Automatic Annotation databases, and their mechanism was studied. A total of 29 compounds and 140 corresponding targets, including 14 key targets and 14 protein factors in protein-protein interaction core network were obtained. The key targets were tumor necrosis factor(TNF), interleukin(IL)-6R and sestrogen receptor alpha. The number of GO items was 466 (P<0.05), including 399 items of biological process (BP), 54 items of cell composition (MF) and 13 items of molecular function (CC). KEGG pathway enrichment screened 85 signaling pathways (P<0.05), including the IL-17 signaling pathway, TNF signaling pathway, advanced glycation end products and their receptors signaling pathway and cAMP signaling pathway. The active ingredients of NZZ. exert their anti-OP effects through multi-components, multi-targets and multi-pathways, which can provide new evidence for further study of their anti-OP mechanism.
Subject(s)
Osteoporosis/pathology , Research/classification , Ligustrum/adverse effects , Genes , Network Pharmacology/instrumentation , Software/classification , Tumor Necrosis Factor-alpha/pharmacology , Glycation End Products, Advanced/adverse effects , Interleukin-17/analogs & derivatives , Gene Ontology , East Asian People , Medicine, Chinese TraditionalABSTRACT
OBJECTIVE@#The present study explored the effects of the combined herbal therapy consisting of curcumin (CUR) and Fructus Ligustri Lucidi (FLL) on aspects of bone regeneration.@*METHODS@#Prior to analyzing the ability of this novel combined herbal therapy to promote aspects of bone regeneration, its cytotoxicity was determined using MC3T3-E1 cells (pre-osteoblast model). Cell proliferation was evaluated using phase-contrast microscopy and cell differentiation was estimated using alkaline phosphatase activity. The effect of the combined herbal therapy (CUR + FLL) was also assessed in terms of mineralization in the extracellular matrix (ECM) of cultured cells. Further, to explore the molecular mechanisms of bone formation, time-dependent expression of bone-regulating protein biomarkers was also evaluated.@*RESULTS@#Combined herbal therapy (CUR + FLL) significantly upregulated the viability, proliferation and differentiation of MC3T3-E1 cells compared to the monotherapy of CUR or FLL. The magnitude of ECM mineralization (calcium deposition) was also higher in MC3T3-E1 cells treated with combined therapy. The time-dependent expression of bone-forming protein biomarkers revealed that the tendency of expression of these bone-regulating proteins was remarkably higher in cells treated with combined therapy.@*CONCLUSION@#The co-administration of CUR and FLL had superior promotion of elements of bone regeneration in cultured cells, thus could be a promising alternative herbal therapy for the management of bone erosive disorders such as osteoporosis.
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Objective: To investigate the protecting and regulatory effects of water extract from Fructus Ligustri Lucidi (FLL) on bone structure and bone metabolism in osteoporosis rats. Methods: SD female rats were bilaterally ovariectomized to establish osteoporosis model, and Sham operated rats only cut the fat around the ovary. Experimental rats were divided into four groups: Sham operation (SHAM) group, model (OVX) group, alendronic acid sodium (ALN) group, and Fructus Ligustri Lucidi (FLL) group, with nine rats in each group. The rats in FLL group were given FLL water extract (3.5 g/kg) and rats in ALL group were given ALN suspension (0.12 g/kg) by ig administration for 12 weeks. At the end of the experiments, the contents of serum and urine calcium (S-Ca) and phosphorus (S-P), urine creatinine (U-Ca/Cr and U-P/Cr), serum high density lipoprotein (HDL-C) cholesterol, low density lipoprotein (LDL-C) cholesterol, total cholesterol (TC), and triglyceride (TG) were measured by biochemical methods. The levels of collagen I amino terminal peptide (PINP), collagen I carboxyl terminal peptide (CTX-I), osteocalcin (OCN), and urine deoxypyridinoline (DPD) were measured with ELISA. The determination of alkaline phosphatase (ALP) was by radioimmunoassay method. To evaluate the change of bone tissue structure, the bone density instrument, VivaCT, and a universal testing machine were used. Results: FLL could inhibit the increased body weight of ovariectomy (OVX) rats, increase S-Ca, S-P, serum HDL and PINP contents, reduce urinary U-Ca/Cr and U-P/Cr ratios, reduce serum LDL-C, TC, TG, ALP, OCN, CTX, and reduce urinary DPD content (P < 0.05 or 0.01) in OVX rats. Meanwhile, FLL can elevate the femur head and vertebral bone mineral density, bone micro-structure and bone strength in OVX rats. Conclusion: FLL can improve the bone density and bone strength in OVX rats by regulating Ca and P metabolism, collagen and non-collagen metabolism.
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This study aimed to trace sources and quantitatively analyze the specnuezhenide content of circular Fructus Ligustri Lucidi for clinical use. Different specifications of Fructus Ligustri Lucidi were identified using DNA barcoding technology and the specnuezhenide content was analyzed by High Performance Liquid Chromatography (HPLC). The ITS sequence of circular Fructus Ligustri Lucidi was identical to that of standard privet, which was determined through botanical identification. ITS sequence similarity between circular Fructus Ligustri Lucidi and Fructus Ligustri Lucidi which was registered in NCBI ranged from 99.5% to 100%. The sequences of circular and other Fructus Ligustri Lucidi were clustered in a Neighbor-Joining tree with bootstrap value of 95, and these sequences could be distinguished from adulterants. Conforming to pharmacopoeia standard, the average specnuezhenide content of circular Fructus Ligustri Lucidi was higher than that of chicken waist Fructus Ligustri Lucidi. Circular Fructus Ligustri Lucidi derived from Ligustrum lucidum Ait. and the specnuezhenide content was higher in circular Fructus Ligustri Lucidi than that in chicken waist Fructus Ligustri Lucidi.
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Chromatography, High Pressure Liquid , DNA Barcoding, Taxonomic , DNA, Plant , Fruit , Chemistry , Glucosides , Ligustrum , Chemistry , Classification , Genetics , Medicine, Chinese Traditional , Polymerase Chain Reaction , Pyrans , Quality ControlABSTRACT
Objective:To explore the effect on sex hormone functional of extracts of Herba Epimedii /Fructus Ligustri Lucidi in osteoporosis rats.Methods:50 cases of 3 months Wistar rats were rando mly divided into normal group ,model group,extract low dose group(extract-L),extract middle dose group(extract-M),extract high dose group(extract-H),10 cases in each group,in addition to the normal group,other 4 groups were bulided OP model by filling and feeding retinoic acid for 15 d,then normal group and model group were filling physiological saline;extract-L,extract-M,extract-H were respectively feeding extracts of Herba Epimedii /Fructus Ligustri Lucidi at 50 mg/kg,100 mg/kg,200 mg/kg,both for 3 weeks.Measured the bone mineral density (BMD),observed the femoral pathology change of left femur with hematoxylin and eosin ( HE ) staining, measured trabecular bone tissue morphology related parameters ,analyzed the hormone receptor protein expression with immunohistochemistry .Results:The BMD of the whole femur ,femur head,and femur neck in normal group were all significantly higher than that of model group ,the BMD of the whole femur ,femur head, and femur neck in extract-M and extract-H group were all significantly higher than that of model group ,the BMD of the whole femur in extract-L group was significantly higher than that of model group ,the differences were statistically significant (P<0.05 or P<0.01).The bone trabecular number had significantly increased ,integrity,continuity improved significantly ,bone histomorphometry such as Tb.Ar, Tb.Th,and Tb.N in model group were decreased ,and Tb.Sp was increased,extract-M and extract-H group had remarkable effect on each index ,while extract-L group only had a remarkable effect on Tb .Ar and Tb.Th ( P<0.05 or P<0.01 ) , compared with model group.The expression of AR , ER in normal groups were significantly higher than that of model group , the expression of AR , ER in extract-H group was significantly higher than that of model group , the differences were statistically significant ( P<0.05 or P<0.01 ) . Conclusion:This study demonstrated that the combination with TFE and TIFL had potential protective effects on osteoporosis rats induced by retinoic acid .The combination with TFE and TIFL is able to increase BMD ,reverse pathological changes of bone tissue ,co-ordinate hormones of the hypothalamic-pituitary-gonadal axis ,and upregulate the expression of sex hormone receptors .
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Fructus Ligustri Lucidi ( FLL) is a traditional Chinese medicine commonly used in clinic. This paper reviews the phar-macological action of FLL and its compounds, especially demon-strates the regulation and mechanism in calcium homeostasis and bone metabolism.
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Objective:To optimize the extraction process of antitumor active ingredients in fructus ligustri lucidi. Methods:Using fructus ligustri lucidi medicinal herb as the raw material, the extraction process of luteolin and pelargidenon in the herb was screened by D-optimal design. The extraction yield of different extraction processes was compared to obtain the optimal extraction conditions. Re-sults:The best extraction conditions were as follows:adding 12-fold 95% ethanol, extracting 2 times with 60 minutes for each time. Conclusion:The optimal extraction technology can provide significant reference for the further pharmacodynamic effect development of fructus ligustri lucidi.
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Objective To study the regulatory effect of ethanol extract of glossy privet fruit and its monomer tyrosol on the adhesion and migration of human epidermal melanocytes.Methods Epidermal melanocytes were isolated from human foreskin,and subjected to a primary culture.Mter 3-5 passages,the melanocytes were treated with various concentrations of ethanol extract of glossy privet fruit (0.0375-0.6 mg/ml)and tyrosol (0.125-2 mmol/L) for 24-72 hours.The XTT colorimetric assay was carried out to evaluate the proliferation of melanocytes,fibronectin (FN)-coated culture plates were used to evaluate the adhesion activity of melanocytes,and Transwell assay was conducted to assess the migration activity of melanocytes.Confocal laser microscopy was utilized to observe the structure and distribution of actin cytoskeleton in melanocytes,and cellular fluorescence intensity was determined by a semi-quantitative analysis.Statistical analysis was carried out by using unpaired t test.Results The adhesion activity of melanocytes to FN was significantly enhanced by the ethanol extract of 0.0375-0.6 mg/ml from glossy privet fruit (P ＜ 0.05 or 0.01),and by tyrosol of 0.5-2 mmol/L (P ＜ 0.05 or 0.01).As XTT assay showed,neither the ethanol extract of 0.15 mg/ml nor tyrosol of 2 mmol/L had cytotoxicity or promotive effect on cell proliferation.Hence,0.15 mg/ml and 2 mmol/L were determined as the working concentrations of ethanol extract and tyrosol respectively.The number of cells migrating through micropore membranes per high-power field (× 200) was 43.7 and 51.0 in melanocytes treated with the ethanol extract of 0.15 mg/ml and tyrosol of 2 mmol/L,respectively,significantly higher than that in untreated melanocytes (20.3,both P ＜ 0.01).Compared with untreated melanocytes,those treated with the ethanol extract of 0.15 mg/ml and those with tyrosol of 2 mmol/L showed higher intracellular fluorescence intensity (P ＜ 0.01) and more stress fiber bundles which congregated inside the cell membrane and around the nuclei.Conclusions The ethanol extract of glossy privet fruit and its monomer tyrosol can promote the adhesion and migration of human melanocytes in vitro,likely by promoting the congregation of actin cytoskeleton in melanocytes.
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A simple,rapid and sensitive liquid chromatography-mass spectrometry（LC-MS）method was developed for the determination of salidroside in rat plasma and study of its pharmacokinetics after oral administration of suspension of Erzhi Wan and Fructus Ligustri lucidi into Wistar rats.Plasma sample of 200 μL was extracted with acetic ether-isopropanol（2∶1）and the extraction was performed on a Kromasil C18 column（150 mm×4.6 mm,5 μm）with the mobile phase of methanol-water（41∶59,v/v）within a run time of 6.0 min.The analyte was monitored with positive electrospray ionization（ESI）by selected ion monitoring（SIM）mode.The target ions were m/z 323.05 for salidroside and m/z 411.05 for internal standard（IS）geniposide.A good linear relationship was obtained over the range of 5.0-500.0 ng/mL and the lower limit of quantification was 5.0 ng/mL.The validated method was successfully applied to the pharmacokinetic study of salidroside in rat plasma after oral administration of suspension of Erzhi Wan and Fructus Ligustri lucidi.
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A simple,rapid and sensitive liquid chromatography-mass spectrometry (LC-MS) method was developed for the determination of salidroside in rat plasma and study of its pharmacokinetics after oral administration of suspension of Erzhi Wan and Fructus Ligustri lucidi into Wistar rats.Plasma sample of 200 μL was extracted with acetic etherisopropanol (2:1) and the extraction was performed on a Kromasil C18 column (150 mm×4.6mm,5 μm) with the mobile phase of methanol-water (41:59,v/v) within a run time of 6.0min.The analyte was monitored with positive electrospray ionization (ESI) by selected ion monitoring (SIM) mode.The target ions were m/z 323.05 for salidroside and m/z 411.05 for internal standard (IS) geniposide.A good linear relationship was obtained over the range of 5.0-500.0 ng/mL and the lower limit of quantification was 5.0 ng/mL.The validated method was successfully applied to the pharmacokinetic study of salidroside in rat plasma after oral administration of suspension of Erzhi Wan and Fructus Ligustri lucidi.
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Objective To study the effects of Fructus ligustri lucidi and its monomers, tyrosol and oleanotic acid, on the migration of mouse melanoblast cell line (NCCmelb4M5). Methods Cultured NCCmelb4M5 cells were treated with Fructus ligustri lucidi (0.0625, 0.125, 0.25, 0.5, 2 mg/mL), tyrosol (0.02, 0.04, 0.08, 0.16, 0.8 mg/mL) and oleanolic acid (0.0625, 0.125, 0.25, 0.5, 2.5 mg/mL), respectively,for 48 hours followed by the detection of cell proliferation with MTT assay. The working concentration of the three drugs was determined according to the results of MTT assay. Scratch and transwell assays were performed to observe the effect of Fructus ligustri lucidi and its monomers at working concentration on the migration of NCCmelb4M5 cells. Results Based on the results of MTT assay, the working concentration of Fructus ligustri lucidi, tyrosol and oleanolic acid was determined at 0.125 mg/mL, 0.08 mg/mL and 0.0625 mg/mL respectively, and at these concentrations, these drugs exhibited a cytotoxity lower than that of absolute alcohol with no obvious stimulation of cell proliferation. Scratch and transwell assay revealed a promoting effect of both Fructus ligustri lucidi and tyrosol on melanoblast migration (P<0.05), while oleanolic acid had little effect on melanoblast migration. Conclusions The extract of Fructus ligustri lucidi has a significant stimulatory effect on the migration of mouse melanoblasts, and tyrosol may be an active component of Fructus ligustri lucidi associated with confirmative effect on migration of mouse melanoblasts.
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Objective To determine oleanolic acid(OA)and ursolic acid(UA)from Fructus Ligustri Lucidi in diffrernt habitat and various growing stages.Method We picked the fruits of Fructus Ligustri Lucidi in Otober in five cities of Shanxi,and in August,September,October,November and December in Xi'an.Removed impurities and storaged the fruits under room temperature.By HPLC with Waters 600 as its chromatographic system,and Lichrospher C_(18)(4.6mmx250mm,5?m) column was applied with CH_3CN-CH_3OH-H_2O-H_3PO_4-(C_2H_5)_3N(50:30:20:0.02:0.04)as its mobile phase,the flow rate was 1 mL/min.The standard working curve was made to determine the contents of OA and UA at different habitat and different time spot from samples.Result The contents of OA and UA were highest in Ankang city.During prolonging growing stages,the highest contents of OA and UA were October and August,respectively.They both reduced to the lowest point in December.Conclusion The contents of OA and UA changed in different habitat and diffrernt growing stages of Fructus Ligustri Lucidi. It was suggested that we should mainly base on the highest contents to select the harvest time according to our demands.
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Objective To study the quality control standard of Gushuling capsule. Methods Fructus Psoraleae and Fructus Ligustri Lucidi in Gushuling capsule were identified by TLC; The content of ieariin was determined by HPLC with acetonitrile-water (30: 70) as mobile phase and the wavelength at 270 nm. Results TLC identification was positive; the linear range of icariin was 0.0968～0.4840 ?g(r=0.9999), the average recovery was 97.27%(RSD=1.88%, n=5). Conclusion This method can be used as a standard of the quality control for Gushuling capsule.
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Objective To establish the HPLC fingerprints of Fructus Ligustri Lucidi.Methods HPLC-ELSD analysis was carried out with Lichrospher C18 column(4.6 mm ? 250 mm,5 ?m).The detector was Alltech ELSD 2000.The method was developed by gradient elution with methanol and water as the mobile phase.With oleanolic acid as the marker,sixteen batches of Fructus Ligustri Lucidi were analyzed with computer-aided similarity evaluation system.Results HPLC fingerprints of Fructus Ligustri Lucidi showed 14 characteristic peaks and the similarity of the fingerprints of 10 batches of samples was over 0.90,indicating the fingerprints being stable and repeatable.Conclusion The method is simple,accurate and with a good reproducibility,and can be used for the identification and quality control of Fructus Ligustri Lucidi.
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Objective To investigate the effect of Fructus Ligustri Lucidi(FLL),a kidney-tonifying Chinese herbal medicine,on calcium metabolism and vitamin D-dependent gene expression in ovariectomized(OVX) rats.Methods Four weeks after OVX surgical operation the OVX rats were ig admini-(stered) with estradiol and FLL for 14 weeks.Ca~(2+) Level in serum,urine,and feces was measured by colorimetric methods,and gene expressions in duodenum and kidney were analyzed by reversed transcriptase-polymerase chain reaction(RT-PCR).Results Ovariectomy of rat led to significant loss of calcium in vivo,as demonstrated by decreasing serum Ca~(2+) level and increasing Ca~(2+) excretion rate in urine and feces in OVX rat group.Treatment of OVX rats with FLL could effectively restore serum Ca~(2+) level and prevent the increase of Ca~(2+) excretion from urine and feces.RT-PCR Analysis showed that both estradiol and FLL could prevent OVX-induced decrease of mRNA expression in duodenal vitamin D receptor(VDR).In kidney,FLL did not alter mRNA expressions of 25-hydroxyvitamin D 1-hydroxylase(1-OHase) and 25-hydroxyvitamin D 24-hydroxylase(24-OHase),while increased the mRNA expression levels of renal cal-ciumbinding protein-9k(CaBP-9k) and calcium-binding protein-28k(CaBP-28k).Conclusion FLL could improve the state of calcium imbalance induced by estrogen deficiency and the potential mechanism that contributes to the improvement of calcium status by FLL might involve the increase in intestinal sensitivity to active vitamin D through its up-regulation of VDR expression in duodenum and the enhancement in Ca~(2+) reabsorption via the up-regulation of calcium binding proteins expression in kidney.
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Objective To optimize the processing technology of Fructus Ligustri Lucidi.Methods A RP-HPLC method was carried out to determine the contents of oleanolic acid and ursolic acid in the fruits of Ligustrum lucidum and an orthogonal design to optimize the processing conditions with the indexes of oleanolic acid content and ursolic acid content.Results Linear ranges of oleanolic acid and ursolic acid were in 0.019 78～0.197 8 mg/mL(r=0.999 8)and in 0.010 26-0.102 6 mg/mL(r=0.999 8)respectively,and average recoveries of oleanolic acid and ursolic acid were 99.53 %(RSD=2.08 %)and 99.88 %(RSD=1.76 %)respectively.Orthogonal design showed the optimum conditions were as follows:soaking for 8 hours with 2 gram wine(5 gram of Fructus Ligustri Lucidi),then steaming for 8 hours.Conclusion HPLC is an accurate and quick method to determinate the contents of oleanolic acid and ursolic acid,and can be used as a content determination method for Fructus Ligustri Lucidi.Soaking time,steaming time and wine content have no significant influences on the contents of oleanolic acid and ursolic acid.
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Objective: To establish the quality standards for Jifukang Oral Solution. Methods: Fructus Corni, Fructus Ligustri Lucidi, Herba Epimedii were identified by TLC, and the content of astragalin A was determined by TLC scanning. Results: The Fructus Corni, Fructus ligustri licidi, Herba Epimedii could be identified by TLC. Astragalin A showed a good linear relationship at a range of 1.065～5.325?g,r=0.9991. The average recovery was 95.1%, and RSD was 2.0%. Conclusion: The methods are accurate and can be used for the quality control of Jifukang Oral Solution.
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The qualitative and quantitative analyses of oleanolic acid in the processed products of Fructus Ligustri Lucidi and the comparative study of its protection action for liver have been carried out. As a result,the product steamed with wine showed the highest oleanolic acid content. Its action of depressing glutamic pyruvic transaminase was also the strongest. These studies provide a base of rational clinical application of the medicine.
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Objective: To study the chemical constituents of Chinese herb Fructus Ligustri lucidi , providing theoretic evidence for its exploitation and utilization. Methods: Chemical constituents of Fructus Ligustri lucidi were isolated and purified by solvent extraction and chromatography; their structures were identified by physiochemical properties and spectroscopic analysis. Results: Seven compounds were isolated from the ethanol extract of the fruits and identified as oleanolic acid(Ⅰ), luteolin 7 O ? D glucoside(Ⅱ), quercitrin(Ⅲ), ? sitosterol 3 O ? D glucoside(Ⅳ), p hydroxyphenethyl ? D glucoside(Ⅴ), D mannitol(Ⅵ) and hexitol(Ⅶ). Compounds Ⅱ,Ⅲ and Ⅶ were obtained from this plant for the first time. Conclusion: Seven compounds have been obtained from the title plant, which provides a base for its future use.