OBJECTIVETo study the chemical constituents of Dolomiaea souliei.

METHODVarious chromatographic techniques were adopted to separate the constituents, and the spectrum analysis was made to identify their structures.

RESULTSeventeen compounds were isolated and identified as: dehydrocostus lactone (1), costunolide (2), mokko lactone (3), santamarine(4), reynosin (5), 4alpha-hydroxy-4beta-methyldihydrocostol (6), sulfocostunolide A (7), beta-costic acid (8), beta-cyclocostunolide (9), vladinol A (10), ursolic acid (11), betulinic acid (12), betulin (13), dibutyl terephthalate (14), dibutyl phthalate (15), uridine (16), and emodin (17).

CONCLUSIONCompounds 6-9 and 12-17 were obtained from this genus for the first time, and compound 11 was obtained from this plant for the first time.

4-Butyrolactone ; analogs & derivatives ; chemistry ; Asteraceae ; chemistry ; Emodin ; chemistry ; Lactones ; chemistry ; Sesquiterpenes ; chemistry ; Triterpenes ; chemistry

Ligustilide is contained highly or around 1% in such umbelliferous plants as Angelica sinensis and Ligusticum chuanxiong, is one of main bioactive constituents. It shows many pharmacological activities related to their efficacy. At present, ligustilide has attracted extensive attention and more and more studies have been reported, indicating that it is a promising compound. This essay summarizes the progress of pharmacological effects of ligustilide on neuroprotection, vasodilatation, anti-caner and anti-tumor, analgesia and anti-inflammation, and pharmacokinetics including absorption, distribution, metabolism and excretion, providing basis for further studies and development of ligustilide.
4-Butyrolactone ; analogs & derivatives ; pharmacokinetics ; pharmacology ; Angelica sinensis ; chemistry ; Animals ; Drugs, Chinese Herbal ; pharmacokinetics ; pharmacology ; Humans ; Ligusticum ; chemistry

OBJECTIVETo compare 39 root samples of Angelica sinensis and A. acutiloba from China and Japan for the quality control of Chinese Angelica.

METHODAn HP-5 (0.32 mm x 30 m, 0.25 microm) column was used for the GC-MS analysis. The oven temperature was programmed from 120 degrees C to 280 degrees C at a rate of 5 degrees C x min(-1). Using principal component analysis, cluster analysis, and discriminant analysis on the sample fingerprints for chemical pattern recognition research.

RESULTZ-Ligustilide was the key principle distinguishing Chin samples from Japan. Moreover, using discriminant analysis, seven samples (four of A. sinensis, three of A. acutiloba) were validated. All samples tested were successfully classified according to their species origin.

4-Butyrolactone ; analogs & derivatives ; chemistry ; Angelica sinensis ; chemistry ; Cluster Analysis ; Discriminant Analysis ; Gas Chromatography-Mass Spectrometry ; methods ; Principal Component Analysis

Bioactivity of Danggui is linked to the content of ligustilide, but the relationship between ligustilide with herb shape, cultivating areas and plant species is still unknown. The relationship was investigated by quantifying on the amounts of Z-ligustilide and E-ligustilide by HPLC-DAD-MS method, and then comparing the content of ligustilides (the sum of Z-ligustilide and E-ligustilide) among forty-four various "Danggui" samples containing thirty Chinese Danggui (CDG), six Japanese Danggui (JDG), four Korea Danggui (KDG) and four European Danggui (EDG). Results showed that the content of ligustilides in CDG samples (Angelica sinensis) was in the range of 5.63-24.53 mg x g(-1) with the mean of 11.02 mg x g(-1) (n = 30). Ligustilides amounts were varied among samples cultivated in different areas in China, i. e. 13.90 mg x g(-1) (n = 6) in Yannan, 12.51 mg x g(-1) (n = 6) in Sichuan and 10.04 mg x g(-1) (n = 13) in Gansu. It was also found that ligustilides content was related to the shape, color and fragrance of herb, e. g. the relative larger amount of ligustilides was in the small main root, long rootlet and perfumed sample. Further, ligustilides contents were estimated to be 1.00 mg x g(-1) (n = 6) in JDG samples (A. acutiloba and A. acutiloba var. sugiyamae) and 2.78 mg x g(-1) (n = 2) in EDG samples (lovage root, Levisticum officinale). However, ligustilides could not be detected in the four KDG samples (A. gigas) and two EDG samples (angelica root, A. archangelica). It has been concluded that ligustilide is significant variant among plant species, which may result in the variety of bioactivity and therapeutic effect.
4-Butyrolactone ; analogs & derivatives ; analysis ; Angelica sinensis ; chemistry ; China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Geography ; Quality Control

AIM: To investigate chemical constituents of Spatholobus suberectus Dunn. METHODS: Isolation and purification were carried out by column chromatographic methods. Compounds were characterized based on their physical characteristics and spectra data. RESULTS: Seventeen compounds were isolated from ethanol extract of S. suberectus. The structures were elucidated as prestegane B (1), (2R, 3R)-buteaspermanol (2), (+)-medioresinol (3), (2R, 3R)-3,7-dihydroxyflavanone (4), benzeneethanol (5), 4, 7, 2'-trihydroxy-4'-methoxyisoflavanol (6), naringenin (7), blumenol A (8), protocatechuic acid ethyl ester (9), liquiritigenin (10), 7, 4'-dihydroxy-8-methoxy-isoflavone (11), 3, 5, 7, 3', 5'-pentahydroxyflavanone (12), protocatechuic acid (13), glycyroside (14), 8-methylretusin-7-O-β-D-glucopyranoside (15), 3, 3', 4', 5, 6, 7, 8-heptahydroxyflavan (16), and dulcisflavan (17). CONCLUSION All compounds are firstly isolated from the title plant and compounds 1, 3 were isolated from the Spatholobus genus for the first time.
4-Butyrolactone ; analogs & derivatives ; chemistry ; isolation & purification ; Fabaceae ; chemistry ; Lignans ; chemistry ; isolation & purification ; Molecular Structure ; Plant Extracts ; chemistry

This study aimed to investigate the effect and mechanism of ligustilide, the main active ingredient in Ligusticum wallichii, on mitochondria fission after PC12 cell injury induced by oxygen and glucose deprivation/reperfusion(OGD/R). In the experiment, an OGD/R model was established in vitro, and PC12 cells were pre-treated with ligustilide for 3 h, and then the cell viability was detected by CCK-8 method. The effect of different concentrations of ligustilide on the morphology of PC12 cells after OGD/R injury was observed under an inverted microscope. Transmission electron microscopy was used to observe the mitochondrial fission of PC12 cells after OGD/R injury. DCFH-DA immunofluorescence staining method was used to detect intracellular reactive oxygen species(ROS) changes. Changes in mitochondria membrane potential(MMP) were detected by flow cytometry. Hochest 33258 was used to observe the apoptosis of PC12 cells. Western blot was used to detect changes in cytochrome C(Cyt C) content in mitochondria and cytoplasm, and mitochondrial fission-related proteins Drp 1 and Fis 1. All results showed that compared with the model group, ligustilide significantly increased the survival rate of PC12 cells and the number of cells. Further experiments showed that ligustilide inhibited the release of ROS and decline of mitochondrial membrane potential in PC12 cells after OGD/R injury. Moreover, ligustilide reduced the release of Cyt C and promoted the expressions of Drp1 and Fis1 in mitochondrial fission proteins. Verification experiments showed that mitochondrial fission inhibitor mdivi-1 decreased cell survival rate and inhibited fission. The results indicated that ligustilide exerted neuro-protective effects by promoting mitochondrial fission and reducing cell damage. It preliminary proves that the mechanism of ligustilide on ischemic brain injury may be related to the promotion of mitochondrial fission and the maintenance of cell homeostasis.
4-Butyrolactone ; analogs & derivatives ; Animals ; Apoptosis ; Cell Survival ; Glucose ; Mitochondria ; Oxygen ; PC12 Cells ; Rats ; Reactive Oxygen Species ; Reperfusion Injury

Mitochondrion, as the main energy-supply organelle, is the key target region that determines neuronal survival and death during ischemia. When an ischemic stroke occurs, timely removal of damaged mitochondria is very important for improving mitochondrial function and repairing nerve damage. This study investigated the effect of ligustilide(LIG), an active ingredient of Chinese medicine, on mitochondrial function and mitophagy based on the oxygen and glucose deprivation/reperfusion(OGD/R)-induced injury model in HT22 cells. By OGD/R-induced injury model was induced in vitro, HT22 cells were pre-treated with LIG for 3 h, and the cell viability was detected by the CCK-8 assay. Immunofluorescence and flow cytometry were used to detect indicators related to mitochondrial function, such as mitochondrial membrane potential, calcium overload, and reactive oxygen species(ROS). Western blot was used to detect the expression of dynamin-related protein 1(Drp1, mitochondrial fission protein) and cleaved caspase-3(apoptotic protein). Immunofluorescence was used to observe the co-localization of the translocase of outer mitochondrial membrane 20(TOMM20, mitochondrial marker) and lysosome-associated membrane protein 2(LAMP2, autophagy marker). The results showed that LIG increased the cell viability of HT22 cells as compared with the conditions in the model group. Furthermore, LIG also inhibited the ROS release, calcium overload, and the decrease in mitochondrial membrane potential in HT22 cells after OGD/R-induced injury, facilitated Drp1 expression, and promoted the co-localization of TOMM20 and LAMP2. The findings indicate that LIG can improve the mitochondrial function after OGD/R-induced injury and promote mitophagy. When mitophagy inhibitor mdivi-1 was administered, the expression of apoptotic protein increased, suggesting that the neuroprotective effect of LIG may be related to the promotion of mitophagy.
4-Butyrolactone/analogs & derivatives* ; Apoptosis ; Calcium/pharmacology* ; Glucose/metabolism* ; Humans ; Mitochondrial Proteins ; Mitophagy ; Reactive Oxygen Species/metabolism* ; Reperfusion Injury/genetics*

OBJECTIVE: To establish the method to analyze γ-hydroxybutyric acid (GHB) and its precursors 1,4-butanediol (1,4-BD) and gamma-butyrolactone (GBL) in urine through LC-MS/MS and provide evidence for related cases. METHODS: GHB-d6 and MOR-d3 were used as the internal standard. The urine sample was separated by LC after protein precipitation with methanol. The electrospray ion source was for ionization. Each compound was detected through multiple-reaction monitoring (MRM) mode. RESULTS: The limits of detection of GHB and its precursors 1,4-BD and GBL were 0.1, 0.1 and 2 μg/mL. The accuracy was 87.6%-98.1%. The intra-day and inter-day precisions were less than 15% and matrix effects were higher than 80%. CONCLUSION The method is high sensitive, simple, rapid, specific and with high reliability. This study has provided technical support and basic data for forensic cases involving GHB.
4-Butyrolactone/urine* ; Butylene Glycols/urine* ; Chromatography, Liquid ; Forensic Sciences ; Humans ; Hydroxybutyrates/urine* ; Mass Spectrometry ; Reproducibility of Results ; Tandem Mass Spectrometry

OBJECTIVETo study the effect of butenolide (BUT) on cultured chondrocytes differentiation and the possible protective effects of selenium (Se).

METHODSEx-vivo cultured chondrocytes were divided into six groups: (1) Control group (without BUT and Se); (2) Se 0.1 microg/ml control group; (3) BUT 0.1 microg/ml group; (4) BUT 1.0 microg/ml group; (5) BUT 5.0 microg/ml group; and (6) BUT 1.0 microg/ml + Se 0.1 microg/ml group. The expression of collagen II (Col II), collagen X (ColX), basic fibroblast growth factor (bFGF), and parathyroid hormone-related peptide (PTHrP) in (or around) chondrocytes in all groups were analyzed by immunohistochemistry.

RESULTSThe expressions of Col II in 1.0 microg/ml BUT group and 5.0 microg/ml BUT group were significantly lower than those in the control group (P < 0.05). The expression of Col II in 1.0 microg/ml BUT + Se group were significantly higher than those in the 1.0 microg/ml BUT group and 5.0 microg/ml BUT group (P < 0.05). The expressions of bFGF and PTHrP of BUT groups were significantly higher than those in the Se and control groups (P < 0.05). No expression of ColX was observed in all groups.

CONCLUSIONBUT can affect the collagen II synthesis of the chondrocytes. Selenium supplementation may play a protective role.

4-Butyrolactone ; analogs & derivatives ; pharmacology ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; cytology ; Humans ; Protective Agents ; pharmacology ; Selenium ; pharmacology ; T-2 Toxin ; toxicity

OBJECTIVETo assay ligustilide content in the herb of Szechwan Lovage Rhizome (Chuanxiong, CX), which is the dried rhizome of Ligusticum chuanxiong in order to assess the quality.

METHODLigustilide was quantitatively analyzed by high-performance liquid chromatography in 21 CX samples. An Alltima C18 column (4.6 mmx 150 mm, 5 microm) was used as the analytical column. The mobile phase consisted of water and acetonitrile (40:60). The flow rate was maintained at 1.0 mL x min(-1) with the column temperature at ambient conditions. The detection wavelength was set at 350 nm.

RESULTThe average content of Z-ligustilide in 21 CX samples was found to be 7.40 +/- 3.54 mg x g(-1)(x +/- s, n = 21). Therefore,the content of Z-ligustilide in CX should not be less than 0.66% (calculated on the dried basis).

CONCLUSIONThe overall analytical procedure is rapid and accuracy which is considered suitable for the quantitative analysis of ligustilide in CX. The amount of ligustilide in CX samples collected from different cultivation areas was obviously different. However, a relatively higher content of ligustilide was generally found in the CX collected from its main cultivated areas.

4-Butyrolactone ; analogs & derivatives ; analysis ; China ; Chromatography, High Pressure Liquid ; methods ; Ecosystem ; Ligusticum ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results ; Rhizome ; chemistry