Comparison of Zhuyao and Douchi Ligustri Lucidi Fructus Based on Quality Investigation of Commercial Samples / 中国实验方剂学杂志

ObjectiveTo investigate the quality of Ligustri Lucidi Fructus in the market， and the moisture， extract， determination of Zhuyao and Douchi Ligustri Lucidi Fructus were compared to increase the utilization rate of Ligustri Lucidi Fructus. MethodThe properties， moisture， total ash， alcohol-soluble extract content and thin layer chromatography （TLC） identification were determined by the methods of Ligustri Lucidi Fructus included in the 2020 edition of Chinese Pharmacopoeia， and high performance liquid chromatography （HPLC） fingerprint and determination of specnuezhenide and salidroside were established with the mobile phase of 0.2% phosphoric acid aqueous solution （A）-acetonitrile （B） （0-70 min， 92%-65%A） for gradient elution， and the detection wavelength of 220 nm at 0-14 min and 225 nm at 14-70 min. The two different characters of Ligustri Lucidi Fructus were comprehensively compared by the above indicators. ResultExcept for one batch which did not meet the requirements due to the quality of harvesting， the other 12 batches of samples all met the requirements of the 2020 edition of Chinese Pharmacopoeia， but there were two different characters. Comparing the two different characters of Ligustri Lucidi Fructus， it is found that the moisture， total ash， extract， salidroside and specnuezhenide contents of Zhuyao samples were 2.22%-5.19%， 3.91%-4.49%， 32.56%-40.95%， 0.073%-0.170% and 1.45%-4.14%， and these values of Douchi samples were 3.57%-5.61%， 3.65%-4.44%， 41.31%-46.70%， 0.041%-0.067% and 3.01%-4.20%， respectively. ConclusionThe contents of extract and specnuezhenide of Douchi Ligustri Lucidi Fructus are mostly higher than those of Zhuyao Ligustri Lucidi Fructus， while the content of salidroside is lower than that of Zhuyao samples， and there are no significant differences in moisture， TLC identification and total ash content. Based on the above research， if the main purpose is to extract salidroside， it is recommended to choose Zhuyao Ligustri Lucidi Fructus. If the main purpose is to use Ligustri Lucidi Fructus as medicine， it is recommended to choose Douchi Ligustri Lucidi Fructus.

Mechanism and experimental verification of "Polygoni Cuspidati Rhizoma et Radix-Ligustri Lucidi Fructus" combination in treatment of acute gouty arthritis based on network pharmacology / 中国中药杂志

Based on network pharmacology, the mechanism of Polygoni Cuspidati Rhizoma et Radix-Ligustri Lucidi Fructus(PL) combination against acute gouty arthritis(AGA) was explored and preliminarily verified by animal experiment. The chemical components and corresponding targets of PL were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). The active components with oral bioavailability(OB)≥30% and drug-likeness(DL)≥0.18 were screened based on literature, and the related protein targets were collected. Then the protein targets were standardized with the help of UniProt database. The AGA-related targets were searched from GeneCards, NCBI, and DrugBank. The common targets of the disease and the medicinals were yielded by FunRich V3, and the protein-protein interaction(PPI) network was constructed to screen the key targets, followed by Gene Ontology(GO) term enrichment analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis of the key targets. Afterwards, some of the key targets were verified by sodium urate crystal-induced AGA mouse model. A total of 25 active components and 287 targets of PL, 811 targets of AGA, and 88 common targets were screened out. PPI network analysis showed that tumor necrosis factor(TNF), interleukin-6(IL-6), and interleukin-1β(IL-1β) may be the core targets of PL in the treatment of AGA. The key targets were mainly involved in 566 GO terms(P<0.05), including multiple biological processes such as inflammatory response and immune response. Moreover, they were related to 116 KEGG pathways and these pathways were involved in inflammation and immunity, mainly including NOD-like receptor signaling pathway and TNF signaling pathway. Animal experiment confirmed that PL can alleviate ankle swelling, improve abnormal gait, and down-regulate the protein expression of TNF-α, IL-6, and IL-1β in AGA mice, indicating that PL can treat AGA through TNF-α, IL-6, and IL-1β and the feasibility of network pharmacology to predict drug targets. This study preliminarily discussed the key targets and biological signaling pathways involved in the treatment of AGA with PL combination, which reflected the multi-pathway and multi-target action characteristics of Chinese medicine. Moreover, this study laid a scientific basis for research on the treatment of AGA with PL combination, as well as the mechanism of action.

Pharmacokinetics of the main components differing between Ligustri Lucidi Fructus and its wine-steamed product / 药学学报

A quantitative analysis method for ten principal components (phenylethanol, iridoids and triterpenes) of raw Ligustri Lucidi Fructus and its wine-steamed product was developed using liquid chromatography tandem triple quadrupole mass spectrometry (LC-QQQ-MS) to study their pharmacokinetic behavior in vivo. The results of methodological investigation were in accord with the criteria of biological analysis. After a single administration to rats of the water extracts of Ligustri Lucidi Fructus and its wine-steamed product, the plasma concentration of each component at different time points was measured and the pharmacokinetic parameters were determined. The AUC0-24 h and Cmax of the phenylethanol components (salidroside, tyrosol, hydroxytyrosol) were the greatest, suggesting that these components are the main pharmacological substances of Ligustri Lucidi Fructus. In addition, the tmax values of the eight major components were even lower with administration of the wine-steamed product, suggesting that these components are rapidly absorbed. However, the tmax values of specnuezhenide and oleanolic acid were greater with administration of the wine-steamed product, indicating that these two components were more slowly absorbed. A secondary peak phenomenon of tyrosol and hydroxytyrosol were observed in two sample groups, whereas the secondary peak phenomenon of salidroside occurred only with the wine-steamed product. This result suggests that the effect of wine-steamed product could persist for a long period. Meanwhile, the relative bioavailability of specnuezhenide and oleanolic acid was greater than 100% with administration of the wine-steamed product, consistent with the Traditional Chinese Medicine theory of the wine-steamed product being more effective than the raw material. The results reveal the different pharmacokinetic parameters and relative bioavailability of each component of Ligustri Lucidi Fructus and its wine-steamed product, and also demonstrate the variation and correlation of various components in vivo and in vitro, providing an experimental basis for the selection of quality control indexes, mechanisms of processing and the metabolic rule in vivo of Ligustri Lucidi Fructus. These experiments were approved by the Ethics Committee of Institute of Basic Theory for Chinese Medicine, China Academy of Chinese Medicine Science.

Qualitative and quantitative analysis on triterpenoids in Ligustri Lucidi Fructus / 中国中药杂志

Triterpenoids are one of the most active constituents in Ligustri Lucidi Fructus, but only oleanolic acid has been mostly studied. In recent years, a growing number of studies have shown that other triterpenes from Ligustri Lucidi Fructus also have various biological activities, so it is necessary to build up a detailed profile of the triterpenoids in Ligustri Lucidi Fructus. The chromatographic separation was performed on a C_(18) column(4.6 mm×250 mm, 5 μm) with mobile phase of methanol-acetonitrile-0.2% formic acid for gradient elution. The detection wavelength was set at 210 nm, with a flow rate of 0.5 mL·min~(-1), and the column temperature of 25 ℃. The HPLC fingerprint of triterpenoids in Ligustri Lucidi Fructus was built by testing 21 batches of samples from different sources. The structures of the total 15 common chromatographic peaks were elucidated with UHPLC-ESI-Orbitrap-MS/MS technique and six of them were identified as tormentic acid, pomolic acid, maslinic acid, botulin, oleanolic acid and ursolic acid by comparison to the reference substances. Under the same chromatographic condition, four main triterpenes(podocarboxylic acid, hawthorn acid, oleanolic acid and ursolic acid) were quantified and the results of system adaptability and methodology investigation all met the requirements of content determination. Meanwhile, with oleanolic acid(A) as the internal reference substance, quantitative analysis of multi-components by single marker(QAMS) method was used to analyze the above four components. The relative correction factor of oleanolic acid(B), hawthorn acid(C) and ursolic acid(D) to oleanolic acid was f_(B/A)=1.12, f_(C/A)=1.02 and f_(D/A)=0.88, respectively, and the relative retention values of these three to oleanolic acid was RRV_(B/A)=0.46, RRV_(C/A)=0.70 and RRV_(D/A)=1.03, respectively. The contents determined by two methods were similar. In conclusion, the method built in this paper is proved to be simple, reliable and specific for the simultaneous qualitative and quantitative analysis of the triterpenoids in Ligustri Lucidi Fructus, which can lay foundation for further assays of the triterpenoids in Ligustri Lucidi Fructus and the relative products.

Comparative study on HPLC fingerprints between crude and processed Ligustri Lucidi Fructus / 中国中药杂志

To establish high performance liquid chromatography(HPLC) fingerprints for crude and processed Ligustri Lucidi Fructus,and to evaluate their quality through the similarity calculation and chemical pattern recognition. The separation was performed with Syncronis C_(18) column(4.6 mm × 250 mm, 5 μm), with acetonitrile(A) and 0.1% phosphoric acid solution(B) as the mobile phase for gradient elution, and a detection wavelength of 280 nm. HPLC was used to detect 22 batches of crude and processed Ligustri Lucidi Fructus,and the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(2012 Edition) was used to evaluate the similarity among 22 batches. The research on pattern recognition was conducted with cluster analysis(CA), principal component analysis(PCA), and partial least squares discriminate analysis(PLS-DA). HPLC fingerprints of crude and processed Ligustri Lucidi Fructus were established, with similarity ranging from 0.9 to 1.0. The crude and processed Ligustri Lucidi Fructus can be obviously distinguished by using CA, PCA and PLS-DA. According to the results of PLS-DA,11 constituents including hydroxytyrosol, tyrosol, specnuezhenide and oleuropein were the main marker components leading to the difference. The established fingerprint method is stable and reliable, and can provide method basis for quality control of crude and processed Ligustri Lucidi Fructus. Chemical pattern recognition is proved to be helpful in comprehensive quality control and evaluation of Ligustri Lucidi Fructus before and after the process.

Molecular Mechanism of Erzhiwan in Inhibiting Benign Prostatic Hyperplasia Base on Network Pharmacology / 中国实验方剂学杂志

Objective: To explain the "multi-components, multi-targets, multi-pathways" mechanism of Erzhiwan in treating benign prostatic hyperplasia(BPH) based the network pharmacology. Method: Ingenuity pathway analysis(IPA) was used to construct components-targets-diseases network and PPI network, then the classified enrichment analysis of gene ontology (GO) and pathway enrichment analysis (KEGG) were carried out on the main function of its gene sets, so as to discuss the mechanism of Erzhiwan in the treatment of BPH. Result: Erzhiwan has 19 components in IPA; and apigenin,luteolin,oleanolic acid and quercitrin were common components of Ligustri Lucidi Fructus and Echiptae Herba and the main component of Erzhiwan. Muscarinic acetylcholine receptor M3 (CHRM3), muscarinic acetylcholine receptor M2 (CHRM2), urokinase plasminogen activator receptor (PLAUR), kinin releasing enzyme 3 (KLK3), cadherin 1 (CDH1), chemokines 3 (CCL3) and metalloproteinase-1 (MMP-1) were important targets for Erzhiwan to treat BPH. The target proteins in PPI network were enriched with 20 GO functions and 5 main KEGG pathways, and Docking was verified for relevant targets. Conclusion: Erzhiwan may play a role in treating BPH by activating MMP-1 and inhibiting KLK3 and CCL3 protein expressions, inducing apoptosis, inhibiting cell proliferation and intervening relevant pathways of mitogen-activated protein kinase/extracellular signal-regulated kinase(MAPK/ERK) and nuclear factor(NF)-κB(NF-κB).

Molecular Mechanism of Ligustri Lucidi Fructus-Astragali Radix for Anti-cancer Based on Network Pharmacology / 中国实验方剂学杂志

Objective: To screen out active ingredients, and predict the anti-cancer targets of Ligustri Lucidi Fructus-Astragali Radix based on the "herbs-active ingredient-action targets" network. Method: The traditional Chinese medicine (TCM) system pharmacology platform (TCMSP) was employed to screen out the active ingredients and putative targets of anti-cancer of glossy privet fruit and astragalus. DisGeNET database and Online Mendelian Inheritance in Man (OMIM database) were employed to predict the targets for treating cancer, and then "herbs-active ingredients-key targets" network was constructed by using Cytoscape software. The omicshare platform was employed to match the putative targets of ingredients and the targets for treating cancer. Finally, the protein interaction network of key targets was constructed by using String database, and the analysis of biological functions and pathways of them was carried out by using DAVID database. Result: Totally 33 drug active ingredients were screened out, involving a total of 203 targets, and 14 of them were related to cancer. These 14 key targets played an therapeutic role in treating cancer by regulating target proteins, such as ERBB2, AR, SRC, EGFR, ESR1, as well as proteoglycans in cancer, cancer pathways, microRNAs in cancer and other pathways. Conclusion: The therapeutic mechanism of Ligustri Lucidi Fructus-Astragali Radix reflects the multi-component, multi-target, and multi-pathway characteristics of TCMs, providing the scientific basis for further study and the material basis of Ligustri Lucidi Fructus-Astragali Radix against cancer.

Effects of Ligustri lucidi Fructus Extract on the Differentiation of Osteoclasts and the Proliferation of Osteoblasts / 中国药房

OBJECTIVE： To investigate the effects of Ligustri lucidi Fructus extract on the differentiation of osteoclasts and the proliferation of osteoblasts. METHODS： Rabbit primary bone marrow cells were induced with 1a，25-Dihydroxyvitamin D3 to obtain osteoclasts. After treated with low-dose， medium-dose and high-dose of L. lucidi Fructus ethanol and water extract （both 2， 20， 200 mg/L）， TRACP activity determined method was used to detect the activity of TRACP in cell lysis solution and the number of TRACP positive cells. The number of celluar bone resorption lacunae and the percentage of bone lacunae area were detected by toluidine blue staining. Osteoblasts were obtained by L-ascorbic acid， β-sodium glycerophosphate and dexamethasone-induced subcloning 14 of cranial parietal anterior osteocytes in mice. MTT assay and APK activity assay were used to detect relative proliferation rate of cells and the activity of APK. RESULTS： After treated with different doses of L. lucidi Fructus extract， the number of TRACP positive cells， the number of bone resorption lacunae and their area were changed in varying degrees. TRACP activity， the number of its positive cells， the number of bone resorption lacunae and the percentage of bone lacunae area in different dosage groups of L. lucidi Fructus ethanol extract and high-dose of water extract； the TRACP activity， the number of its positive cells and the percentage of bone lacunae area in L. lucidi Fructus water extract medium-dose group were decreased significantly， while the number of bone resorption lacunae in L. lucidi Fructus water extract low-dose group was increased significantly （P＜0.05 or P＜0.01）. The relative proliferation rate of cells in L. lucidi Fructus extract low-dose and medium-dose groups and APK activity of cells in extract groups were significantly increased， while the relative proliferation rate of cells were decreased significantly in L. lucidi Fructus ethanol extract and water extract high-dose groups （P＜0.01）. CONCLUSIONS： L. lucidi Fructus extract can inhibit TRACP activity of osteoclasts， change the bone resorption function of osteoclasts and the proliferation behavior of osteoblasts and increase APK activity of osteoclasts.

Qualitative and quantitative analysis on non-triterpenoids in Ligustri Lucidi Fructus / 中国中药杂志

In order to improve the quality control level of Ligustri Lucidi Fructus(LLF) and to explore the changes of chemical components after processing,the HPLC method for fingerprint and simultaneous determination of the major polar components in LLF were established. The octadecylsilane bonded silica gel was used as the stationary phase,with acetonitrile as the mobile phase A and0. 2% formic acid as the mobile phase B in a gradient elution procedure at a flow rate of 1. 0 m L·min-1. The detection wavelength was set at 280 nm and the column temperature was 25 ℃. There were 22 common peaks,20 of which were selected from the fingerprint of LLF and its wine-steamed product,respectively,and 14 chromatographic peaks were identified with reference substances. With the same chromatographic conditions,seven components were quantitatively analyzed and the results of system adaptability and methodology investigation all met the requirements of content determination. Compared with the crude LLF,the content of 5-hydroxymethyl furfural and salidroside significantly increased in wine-steamed LLF,while the contents of iridoid glycosides generally decreased. The method provided a basis for quality control of LLF and its processed products as well as the related preparations.

Study on quality characteristics of Ligustri Lucidi Fructus in national resource survey and consideration on standards of Ligustri Lucidi Fructus in Chinese Pharmacopoeia / 中国中药杂志

The content of tyrosol,salidroside,echinacoside,rutin,acteoside,ligustroflavone,specnuezhenide,and quercetin were determined by HPLC,and the color of Ligustri Lucidi Fructus was determined by comparison with color card.Hundred-seed weight was analyzed by using gravimetric method.The correlation analysis and One-way ANOVA were used to analyze the relationship between the characters,the chemical composition,the harvest time and the geographical location of Ligustri Lucidi Fructus,for giving a comprehensive evaluation of the quality of Ligustri Lucidi Fructus The results showed that 92% of Ligustri Lucidi Fructus were all up to quality standard of Chinese Pharmacopoeia,and the contents of 7 components in Ligustri Lucidi Fructus(except quercetin) were higher than those in samples with black colors.The content of salidroside in Ligustri Lucidi Fructus harvested in June was the highest and the other7 components of Ligustri Lucidi Fructus were relatively high in 8-10 months.According to the quality parameters of Ligustri Lucidi Fructus,the Ligustri Lucidi Fructus from six habitats can not be distinguished effectively.The results showed that there was a certain relationship between the color,harvest season and component content of Ligustri Lucidi Fructus,and the habitats were not related to the quality parameters of Ligustri Lucidi Fructus.The study aimsat providing data support for the resource status of native Ligustri Lucidi Fructus,and a theoretical basis for the revision of standards of Ligustri Lucidi Fructusin the future.

Characterization of co-amorphous complex with piperine and triterpenoids from Ligustri Lucidi Fructus and dissolution evaluation in vitro / 中草药

Objective To prepare a co-amorphous complex of piperine (PIP) and triterpenoids from Ligustri Lucidi Fructus (TLLF) to improve the dissolution of TLLF. Methods TLLF-PIP co-amorphous complex was prepared using solvent evaporation method. The microscopic structure of co-amorphous complex (CAC) was analyzed using differential scanning calorimetry (DSC), scanning electronmicroscopy (SEM), and powder X-ray diffraction (PXRD), the optimal ratio of the two components was investigated through the thermal stability experiment. Ultraviolet-visible spectrophotometry (UV-Vis) and high performance liquid chromatography (HPLC) were applied to determine the amount of TLLF, PIP and the individual components, ursolic acid (UA) and oleanolic acid (OA), respectively. The interaction of the optimal CAC was analyzed by Fourier transform infrared spectroscopy (FTIR), and its solubility and dissolution behavior in vitro were also studied to evaluate the formation of the preparation. Results PXRD analysis indicated that the TLLF-PIP-CAC with a mass ratio of 1:1 exhibited a stable amorphous state, which particles presented in near-spherical shape. FTIR results indicated that there are some hydrogen interactions between the moleculars of TLLF and PIP. The solubility and dissolution determination exhibited a pairwise released behavior with improved triterpenoids and decreased piperine dissolution. Conclusion The prepared TLLF-PIP-CAC can significantly improve the dissolution and amorphous stability of TLLF, which may provide a reference for the compatibility of insoluble compound in traditional Chinese medicine.

Content change of main components in processed Ligustrum lucidum fruits of different processing time and their characteristic quality standard based on UPLC technology / 中国中药杂志

To investigate the dynamic change law of the main components in Ligustri Lucidi Fructus during the wine-steaming process and attempt to establish the characteristic quality standard of wine-steamed Ligustri Lucidi Fructus by determining the content of salidroside and specnuezhenide using Ultra Performance Liquid Chromatography (UPLC) technology at different processing time points (12, 15, 18, 21, 24 h). The chromatographic separation was performed on Waters Acquity UPLC®BEH C₁₈ column (2.1 mm×50 mm, 1.7 μm) with acetonitrile and 0.2% formic acid aqueous solution as the mobile phase for gradient elution; and the detection wavelength was set at 280 nm; the flow rate was 0.5 mL·min⁻¹, and the column temperature was set at 40 °C. The results showed that the two components were well separated in the above conditions. The salidroside and specnuezhenide showed a good linear relationship within the range of 10.19-326 ng and 49.53-1 585 ng, respectively. Their average recovery was 103.4% and 101.7% with RSD of 0.81% and 0.79%, respectively. With the extension of processing time, the content of specnuezhenide was decreased, while salidroside was gradually increased. For the 27 batches of Ligustri Lucidi Fructus, the content of salidroside was between 0.042 5% and 0.192 4%, and that of specnuezhenide was between 0.829 7% and 5.218 0%. While for the 25 batches of wine-steamed Ligustri Lucidi Fructus, the content of the first one was between 0.229 2% and 1.045 8%, and the latter one was between 0.743 8% and 3.645 4%. As compared with Ligustri Lucidi Fructus, the ratio of specnuezhenide to salidroside was significantly decreased in the wine-steamed Ligustri Lucidi Fructus. According to the experimental results, the quality standard of Ligustri Lucidi Fructus is tentatively fixed as follows: the content of specnuezhenide shall not be less than 0.80%, and the ratio of specnuezhenide content/salidroside content (Sp/Sa) should not be smaller than 15. As for the wine-steamed ones, the content of salidroside should not be less than 0.20%, and specnuezhenide content should not be less than 0.70%; Sp/Sa should not be greater than 8. The method established in this study is simple and reliable, which could be used for the content detection of salidroside and specnuezhenide in Ligustri Lucidi Fructus samples. The characteristic quality standard established in this study could be used to distinguish the Ligustri Lucidi Fructus and wine-steamed Ligustri Lucidi Fructus.

Dynamic changes of five constituents in Ligustri lucidi Fructus at five picking time / 中成药

AIM To analyze the dynamic changes of five constituents in Ligustri lucidi Fructus at five picking time (August,September,October,November,December).METHODS The HPLC analysis of Ligustri lucidi Fructus ethanol extract was performed on a 25 ℃ thermostatic Aglient Zorbax SB-C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-0.1％ phosphoric acid flowing at 1 mL/min in a gradient elution manner,and the detection wavelength was set at 224 nm.RESULTS Salidroside,tyrosol,luteolin-7-O-glucoside,ligustroflavone and specnuezhenide showed good linear relationships within their own ranges (r ＞0.999 0),whose average recoveries were 99.56％-100.30％ with the RSDs of 0.89％-1.23％.The contents of various constituents (except for tyrosol) were the highest in samples picked up in September,followed by those picked up in October.CONCLUSION The suitable picking time of Ligustri lucidi Fructus is September and October.

Research advances on pharmacologic effects of Ligustri Lucidi Fructus and its active constituents in skeletal muscle / 药物评价研究

Ligustri Lucidi Fructus and its active constituents,ursolic acid and salidroside,have insulin-like effect to produce positive action on skeletal muscle:,It can improve the utilization of carbohydrate and lipid in muscle,increase synthesis of muscular glycogen and protein,muscular weigh,size of fast-and slow-muscle fibers,and accelerate transformation fast-muscle fiber toward slow-muscle fiber.Ursolic acid stimulates muscle satellite cells proliferation,myogenic differentiation of myoblasts,enhances neomyogenesis in skeletal muscle,and promotes skeletal muscle rejuvenation.In addition,Ligustri Lucidi Fructus and its active constituents down-regulating repression of atrogin-1 and muscle ring-finger protein-1 (MuRF-1),and anti-oxidation and anti-inflammation,it may protect and treat for muscle atrophy and fatigue-induced muscle injury.

Quality stability of Ligustri Lucidi Fructus / 中国药学杂志

OBJECTIVE: To study the change rule of specnuezhenide content and antoxidant activity of Ligustri Lucidi Fructus with temperature and time, and determine its validity period. METHODS: Dynamic parameters were established under constant temperature, and specnuezhenide content of Ligustri Lucidi Fructus was determined by HPLC after accelerated test by classical constant temperature method. The shelf life of Ligustri Lucidi Fructus at 25℃ were determined by Arrhenius theory. The antioxidant activities of different samples of Ligustri Lucidi Fructus were evaluated by ABTS assay. RESULTS: The shelf life of Ligustri Lucidi Fructus was forecasted to be 1.83 years at 25℃. The antioxidant activitiy of Ligustri Lucidi Fructus showed a trend of decline after rising first. CONCLUSION: High temperature and storage time would affect the chemical composition of Ligustri Lucidi Fructus, so Ligustri Lucidi Fructus should be kept in the shade for no more than 1.83 years for a good therapeutic effect.

HPLC fingerprint and multi-component content determination of Ligustri lucidi Fructus / 中草药

Objective To establish the HPLC fingerpint method for simultaneous determination of four representative components (salidroside, echinacoside, specnuezhenide and oleuropein) of Ligustri Lucidi Fructus, so as to provide a reference for the quality control. Methods The method was performed on an Diamonsil C18 anlytical column (250 mm × 4.6 mm, 5 μm) at the column temperature of 30 ℃. The gradient mobile phase consisted of acetonitrile (A)-0.1% formic acid (B) with a flow rate of 1.0 mL/min. The detection wavelengths were 224 nm. HPLC-Q/TOF-MS were used for identifing the common peaks. Results By studying comparatively the fingerprints of 11 samples, 14 common peaks have been confirmed. There were 12 common peaks were identified by HPLC-Q/TOF-MSE. The similarity of 10 batches are greater than 0.990. Salidroside, echinacoside, specnuezhenide and oleuropein were baseline separated with good linearity relationships (r > 0.999) between concentration and peak areas over the linear ranges. The average recoveries of the investigated compounds were 97.40%, 98.99%, 97.03%, and 100.55%, respectively. Conclusion The method is accurate, reliable and with good reproducibility. It could be used for quality control and evaluation of Ligustri Lucidi Fructus.

Effect of Ligustri Lucidi Fructus on hepatic COX activity in normal rats and preliminary investigation of its DNA methylation mechanism / 中国中药杂志

The study aims to investigate the effect of Ligustri Lucidi Fructus on cytochrome c oxidase(COX) activity in rat hepatic tissues and explore its possible mechanism of DNA methylation. Male SD rats received aqueous extract of Ligustri Lucidi Fructus (2.0，6.0 g•kg⁻¹) by intragastric administration for 30 d. After the rats were sacrificed, hepatic tissues of rats were taken to detect COX activity, protein concentration of COX4I1, TET1 and DNMT3A protein levels, mRNA expression levels of Cox4i1, Dnmt3a and Tet1, and determine the DNA methylation frequency of Cox4i1.Results showed that both low and high doses of Ligustri Lucidi Fructus could significantly increase COX activity and concentration of COX4I1(P<0.05), with a decreasing tendency of both TET1 protein and DNMT3a protein expression； however, mRNA expression levels of Cox4i1, Dnmt3a and Tet1 were not significantly changed by Ligustri Lucidi Fructus. In addition, DNA methylation frequency of Cox4i1 in high dose group showed a declining tendency as compared with the blank control group, but without significant difference.These results indicated that Ligustri Lucidi Fructus had promotive effect on hepatic COX activity in rats, which may be achieved by increasing protein content of COX4I1. Moreover, a decreased tendency of DNMT3A protein could be one of the reasons for the lower trend of Cox4i1 methylation rate. In addition, Ligustri Lucidi Fructus may regulate the expression levels of DNMT3A and TET1 in the same direction and its mechanism is not clear.

Research progress on effects and their mechanism of Ligustri Lucidi Fructus in treatment of osteoporosis / 中草药

Ligustri Lucidi Fructus (LLF), which could nourish the liver and kidney, was used for the treatment of bone atrophy in ancient times. The current basic and clinical studies have shown that LLF had a broad application prospect in the treatment of osteoporosis. In this paper, by reviewing the related literature of home and abroad, we summarize the related experimental and clinical research progress of LLF and its compound on the treatment of osteoporosis, and clear their function mechanism, hoping to lay a theoretical foundation for further experimental study and clinical research.

HPLC fingerprints of crude and processed Ligustri Lucidi Fructus / 中草药

Objective: To set up an analysis method of chromatographic fingerprint for crude and processed Ligustri Lucidi Fructus by HPLC. Methods: Purospher® STAR RP18 Endcapped column (250 mm×4.6 mm, 5 μm) was used for separation. The separation process was carried out using a gradient elution, which was based on a mobile phase of the mixture aqueous solution with water-acetonitrile at the flow rate of 1.0 mL/min. The detection wavelength was 282 nm and the column temperature was 25℃. The chromatographic fingerprints were analyzed with the Similarity Evaluation System for Chromatographic Fingerprint of TCM (2012. 130723 Edition). Results: Totally 19 common fingerprint peaks and five constituents including salidroside, privet glycosides, especially privet glycoside, oleuropein, and oleonuezhenide were identified after optimization of the detection wavelength, mobile phase, and gradient elution conditions. Furthermore, the similarity of HPLC fingerprints of 10 batches of Ligustri Lucidi Fructus was high, being over 0.9. Conclusion: The present methodology meets the technical requirements of fingerprint with the rapid, simple, and accuracy properties and could be a basis for the intrinsic quality control standard for Ligustri Lucidi Fructus.

Optimization of processing technology of Ligustri Lucidi Fructus stewed with wine based on analytic hierarchy process and multi-index orthogonal test / 中草药

Objective: To optimize the processing technology of Ligustri Lucidi Fructus (LLF) stewed with wine. Methods: The contents of salidroside, specnuezhenide, and oleanolic acid were simultaneously determined using HPLC. The weight coefficient of each component was evaluated by the analytic hierarchy process. With composite score as index, an orthogonal test was adopted to investigate the effects of wine amount, soaking time, and steaming time on the quality of the processed products and to optimize the technical parameters for the processing of LLF stewed with wine. Results: Wine amount and steaming time had significant influence on the quality of products, but soaking time had no significant effect. Optimal processing parameters were as follows: 30 g of wine was added to 100 g of herbs, soaking time was 0.5 h, and steaming time was 8 h. Conclusion: The optimized processing technology is reasonable and reliable, and it can provide a reference for the processing of LLF. The method established to simultaneously determine the contents of three components in LLF is simple, rapid, and reproducible for controlling the quality of LLF.