Eight butyl-phthalides, senkyunolide K(1), senkyunolide N(2), butylphthalide(3), senkyunolide I(4), senkyunolide H(5),(Z)-butylidenephthalide(6),(Z)-ligustilide(7), and 3-butylidene-7-hydroxyphthalide(8) were isolated from the aerial part of Ligusticum sinense by column chromatography on silica gel column, ODS, Sephadex LH-20 and semi-preparative HPLC. Their structures were elucidated on the basis of spectroscopic and chemical data, especially NMR and MS. Compound 1 was a new butyl-phthalide and compounds 2-8 were isolated from the aerial part of L. sinense for the first time. Furthermore, the inhibitory activities of compounds 1-8 against the nitric oxide(NO) production induced by lipopolysaccharide(LPS) in mouse RAW264.7 macrophages in vitro were evaluated. The results showed that compounds 1-8 exerted inhibitory activities on NO production with IC_(50) of 19.34-42.16 μmol·L~(-1).
Animals ; Mice ; Nitric Oxide/biosynthesis* ; Ligusticum/chemistry* ; Benzofurans/isolation & purification* ; Drugs, Chinese Herbal/isolation & purification* ; Macrophages/immunology* ; RAW 264.7 Cells ; Molecular Structure

Ligustilide, a phthalide compound extracted from Umbelliferae plants such as Angelica sinensis and Ligusticum chuanxiong, has been proven to possess various pharmacological activities, such as anti-inflammatory, anti-tumor, anti-atherosclerosis, anti-ischemic stroke injury, and anti-Alzheimer's disease properties. In recent years, it has shown great potential, particularly in the treatment of locomotor system diseases. Studies have shown that ligustilide has significant therapeutic effects on various locomotor system diseases, including osteoporosis, osteoarthritis, femoral head necrosis, osteosarcoma, and muscle aging and injury. Its mechanisms of action include enhancing the differentiation ability of osteoblasts(OBs), inhibiting the formation ability of osteoclasts(OCs), downregulating inflammatory factors, promoting the synthesis of extracellular matrix(ECM), improving local blood supply to the femoral head, balancing lipid metabolism, inhibiting the proliferation and migration of osteosarcoma cells, inducing cell cycle arrest, enhancing glucose utilization in skeletal muscle, and regulating autophagy and apoptosis. However, its clinical application is severely limited by drawbacks such as structural instability, poor water solubility, and low bioavailability. Currently, formulation techniques such as dripping pills, micropills, inclusion complexes, and liposomes are being used to improve its stability and water solubility, thereby enhancing its therapeutic efficacy. This article summarized the effects, mechanisms of action, and pharmacokinetics of ligustilide monomers and preparations in the treatment of locomotor system diseases in China and abroad in recent years, aiming to provide reference and guidance for further development and application of ligustilide in this field.
Humans ; 4-Butyrolactone/pharmacology* ; Animals ; Drugs, Chinese Herbal/chemistry* ; Angelica sinensis/chemistry* ; Osteoporosis/metabolism* ; Ligusticum/chemistry*

Aiming at the phenomenon of heavy metal Cd exceeding the standard of Chuanxiong medicinal materials,the accumulation of 12 inorganic elements,including heavy metals,in Ligusticum chuanxiong was studied in this paper. It was found that the contents and distribution of most inorganic elements in the stems and leaves of L. chuanxiong were higher than those in the rhizomes at seedling and shooting stages. The content of most elements in rhizome reached the highest at harvest stage,and the distribution ratio of some elements in rhizome was higher than that in stem and leaf at harvest stage. But rhizome,stem and leaf of L. chuanxiong have relatively stable absorption capacity and enrichment effect on different elements,and are less affected by growth period and position. Rhizomes and stems and leaves of L. chuanxiong were enriched with Cd,and stems and leaves also accumulated Pb at seedling stage and stem stage. The absorption capacity of Pb in stems and leaves of L. chuanxiong was higher than that of rhizomes,and the ability of absorbing Cd was less than that of rhizomes at harvest time. The total uptake of Cd and Pb by L. chuanxiong decreased with the prolongation of growth time,but the proportion of Cd and Pb in rhizome increased,so that the content of Cd and Pb increased with the prolongation of growth time.
Cadmium ; analysis ; Drugs, Chinese Herbal ; chemistry ; Ligusticum ; chemistry ; Metals, Heavy ; analysis ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Rhizome ; chemistry

Quality constant is a kind of grading evaluation method of traditional Chinese medicine pieces based on the combination of traditional knowledge and modern quality control. This method has been successfully applied in the grading evaluation of Glycyrrizae Radix et Rhizome pieces and traditional Chinese medicine pieces,and with a positive influence in the industry. With Dachuanxiong Formula for example,different grades of Ligusticum chuanxiong and Gastrodia elata pieces formed high-quality,moderate and qualified Dachuanxiong Formula on the basis of the grading evaluation of the pieces,and the pharmacodynamics method was used to evaluate its efficacy. The results showed that the maximum vascular diastolic rates of Dachuanxiong Formula in the three grades were( 80. 3±5. 2) %,( 67. 0±6. 1) %and( 60. 3±6. 5) %,and the strength of pharmacodynamics was positively correlated with the grade of L. chuanxiong and G. elata pieces.The quality constant technique can objectively and quantitatively classify single decoctions,and has important correlations and prompts for the efficacy of traditional Chinese medicine prescriptions composed of these pieces,with important significance in promoting hierarchical management of the industry,implementing better price for high quality and defining high quality and superior effect.
Drugs, Chinese Herbal ; standards ; Gastrodia ; chemistry ; Glycyrrhiza ; chemistry ; Ligusticum ; chemistry ; Medicine, Chinese Traditional ; Plant Roots ; chemistry ; Quality Control ; Rhizome ; chemistry

The method of classifying the quality grade of traditional Chinese medicine slices with cross section model quality constant was applied to the grade evaluation of Ligusticum chuanxiong pieces,and a reasonable grade standard of L. chuanxiong pieces was established. The purpose is to classify the 15 batches of L. chuanxiong pieces by combining the advantages of traditional grading with modern quality control indicators. By measuring the natural morphological parameters,processing parameters and the intrinsic content of ferulic acid,an important active ingredient,of the 15 batches of L. chuanxiong pieces collected from different manufacturers and different batches of different medicinal materials markets,we can synthesize the results. The mass constants and percentage mass constants are calculated and analyzed based on the above data. The results showed that the quality constants of 15 batches of L. chuanxiong pieces collected ranged from 0.53-3.00; if the percentage mass constants were more than 80%,50%-80% was second-class pieces,and the rest were third-class pieces,the evaluation results were as follows: the quality constants of first-class L. chuanxiong pieces were more than 2.40,the quality constants of second-class L. chuanxiong pieces should be 1.70-2.40,and the quality constants of third-class L. chuanxiong pieces should be less than 1.70. In this paper,the method of dividing the quality constants of the top blade model into different grades is further applied and practiced,which proves that the method is scientific,reasonable and multi-adaptable. At the same time,it enriches the research data of the grade evaluation of L. chuanxiong pieces,provides a useful reference for the promotion of the grade evaluation of L. chuanxiong pieces,and lays an experimental foundation for the next research of the subject group.
Drugs, Chinese Herbal ; standards ; Ligusticum ; chemistry ; Medicine, Chinese Traditional ; Quality Control

Ligustrazine is an important active ingredient of the traditional Chinese medicine Chuanxiong Rhizoma, but its content is a controversial topic. The endophytes of medicinal plants have the ability to produce the same active substances as the host, so this report focused on the endophytic Bacillus subtilis, to study the origin of ligustrazine in Chuanxiong Rhizoma preliminarily by inoculating the isolated endophytic B. subtilis to the Chuanxiong Rhizoma medium for solid state fermentation. Tissue grinding method was used to isolate the endogenetic B. subtilis. The morphological features, conventional physiological and biochemical reactions and 16S rRNA molecular techniques were combined to identify the endogenetic strains. Then, the strains that grew well in the medicinal matrix of Chuanxiong Rhizoma were screened out for further fermentation studies. The solid-state fermentation was performed at 37 °C for 30 d using Chuanxiong Rhizoma fermentation medium (40 g Chuanxiong Rhizoma powder, 100 mL sterile water, 121 °C, sterilization for 25 minutes). UPLC was used to detect the contents of ligustrazine, acetoin in the Chuanxiong Rhizoma fermentation medium and Chuanxiong Rhizoma. All the five strains were Gram-positive and had spores. Phylogenetic analysis of the 16S rRNA sequence showed that the endophytes were B. subtilis. The results of UPLC showed that ligustrazine was detected in the Chuanxiong Rhizoma fermentation medium inoculated with endogenetic B. subtilis LB3, LB3-2-1, LB4, LB5 and LB6-2, while not detected neither in blank Chuanxiong Rhizoma fermentation medium nor in Chuanxiong Rhizoma. This study showed that the endogenetic B. subtilis of Ligusticum chuanxiong Hort. can make use of Chuanxiong Rhizoma fermentation medium to produce ligustrazine. Endogenetic B. subtilis has a certain correlation with the accumulation of ligustrazine in Rhizoma Chuanxiong. We speculate that the ligustrazine may be derived from the catabolism of endogenetic B. subtilis in Ligusticum chuanxiong.
Bacillus subtilis ; Endophytes ; Fermentation ; Ligusticum ; chemistry ; microbiology ; Phylogeny ; Pyrazines ; analysis ; RNA, Ribosomal, 16S ; Rhizome ; chemistry

To discuss the availability of evaluation on the dissolution studies of the multicomponents in traditional Chinese medicine, the in vitro dissolution of total composition of the tablet of rhizomes of Ligusticum chuanxiong components and its correlation with the in vivo were studied by the method of area under the absorbance-wavelength curve (AUAWC). Taken the tablet of rhizomes of Ligusticum chuanxiong components which is composed of sodium ferulate and ligustrazine hydrochloride as subject model, the dissolution tests were carried out with basket method. The plasma concentrations of tablets in different rats were determined by AUAWC at different interval times. The in vivo absorption percentage was calculated by Wagner-Nelson equation to evaluate the in vitro and in vivo correlation. According to the results, the cumulative dissolution in vitro of total composition of tablets of rhizomes of Ligusticum chuanxiong components at 60 min was 90.65% in water by AUAWC. The in vivo pharmacokinetics is fitted with an one-compartment model. The linear equation based on the cumulative dissolution rate (fr) and absorption percentage (fa) at 5, 10, 20, 30 and 60 min was fa = 0.819 7 fr+0.183 and the correlation coefficient was 0.959 5, which showed a good correlation between the in vitro dissolution and the in vivo absorption percentage. The method of AUAWC can be used accurately, feasibly and conveniently to evaluate the in vitro and in vivo correlation of total composition of tablets of rhizomes of Ligusticum chuanxiong components, which will provide better guidance to study the in vitro and in vivo correlation of sustained release preparation etc under complex system of traditional Chinese medicine in the future.
Animals ; Coumaric Acids ; chemistry ; Delayed-Action Preparations ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Ligusticum ; chemistry ; Rats ; Rhizome ; chemistry ; Solubility ; Tablets

The study was using the orthogonal test and making the extraction rates of icariin, ferulic acid, epimedin A, epimedin B, epimedin C, baohuoside I and ligustilide determinated by HPLC multiwavelength switch, gradient elution and multi-index comprehensive weighted scoring method (weight coefficient was 0.47: 0.16: 0.07: 0.07: 0.08: 0.06: 0.09) as evaluation index, combine with SPSS 16.0 software to optimizing the best extraction. It was Yinpian soak 1 h, 12 times more than the volumn of 50% ethanol solution, by heating reflux extraction for 60 min. The compliance test indicates that the optimized compatibility extraction technology is stable and practical, and it has provided an experimental basis for compound preparation technology research of Epimedium brevicornu and Ligusticum chuanxiong.
Chemical Fractionation ; methods ; Chemistry, Pharmaceutical ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Epimedium ; chemistry ; Ligusticum ; chemistry

To explore the effective ingredients and mechanism of Ligusticum wallichii in treating brain ischemia. Four brain ischemia-related target proteins were selected in the joint screening for the 45 component in L. wallichii reported in literatures based on molecular docking by reference to the corresponding drugs in the market. According to the docking results, multiple components in L. wallichii, such as phthalides, were superior to the corresponding drugs in the market, suggesting that they may be the major effective components in L. wallichii for treating brain ischemia. The method can be used to study the material base and molecular mechanism of traditional Chinese medicines.
Brain Ischemia ; drug therapy ; Ligusticum ; chemistry ; Medicine, Chinese Traditional ; Molecular Docking Simulation ; Nitric Oxide Synthase Type II ; antagonists & inhibitors ; Peroxidase ; antagonists & inhibitors ; Phytotherapy ; Tumor Necrosis Factor-alpha ; antagonists & inhibitors

The metabolic characteristics of ligustrazin (TMPz) in liver microsomes were investigated in the present study. The reaction phenotyping of TMPz metabolism was also identified by in vitro assessment using recombinant human cytochrome P450 enzymes (CYP) and UDP glucuronosyltransferases (UGT). TMPz was incubated at 37 degrees C with human (HLM) and rat liver microsomes (RLM) in the presence of different co-factors. The metabolic stability and enzyme kinetics of TMPz were studied by determining its remaining concentrations with a LC-MS/MS method. TMPz was only metabolically eliminated in the microsomes with NADPH or NADPH+UDPGA. In the HLM and RLM with NADPH+UDPGA, t1/2, K(m) and V(max) of TMPz were 94.24 +/- 4.53 and 105.07 +/- 9.44 min, 22.74 +/- 1.89 and 33.09 +/- 2.74 micromol x L(-1), 253.50 +/- 10.06 and 190.40 +/- 8.35 nmol x min(-1) x mg(-1) (protein), respectively. TMPz showed a slightly higher metabolic rate in HLM than that in RLM. Its primary oxidative metabolites, 2-hydroxymethyl-3, 5, 6-trimethylpyrazine (HTMP), could undergo glucuronide conjugation. The CYP reaction phenotyping of TMPz metabolism was identified using a panel of recombinant CYP isoforms (rCYP) and specific CYP inhibitors in HLM. CYP1A2, 2C9 and 3A4 were found to be the major CYP isoforms involved in TMPz metabolism. Their individual contributions were assessed b) using the method of the total normalized rate to be 19.32%, 27.79% and 52.90%, respectively. It was observed that these CYP isoforms mediated the formation of HTMP in rCYP incubation. The UGT reaction phenotyping of HTMP glucuronidation was also investigated preliminarily by using a panel of 6 UGT isoforms (rUGT). UGT1A1, 1A4 and 1A6 were the predominant isoforms mediated the HTMP glucuronidation. The results above indicate that the metabolism of TMPz involves multiple enzymes mediated phase I and phase II reactions.
Animals ; Cytochrome P-450 CYP1A2 ; metabolism ; Cytochrome P-450 CYP2C9 ; metabolism ; Cytochrome P-450 CYP3A ; metabolism ; Cytochrome P-450 Enzyme Inhibitors ; Cytochrome P-450 Enzyme System ; metabolism ; Drug Interactions ; Glucuronosyltransferase ; metabolism ; Humans ; Ligusticum ; chemistry ; Microsomes, Liver ; enzymology ; NADP ; metabolism ; pharmacology ; Pyrazines ; metabolism ; pharmacokinetics ; Rats ; Uridine Diphosphate Glucuronic Acid ; metabolism ; pharmacology