Dysregulated Gln-Glu-α-ketoglutarate axis impairs maternal decidualization and increases the risk of recurrent spontaneous miscarriage.

Recurrent spontaneous miscarriage (RSM) affects 1%-2% of fertile women worldwide and poses a risk of future pregnancy complications. Increasing evidence has indicated that defective endometrial stromal decidualization is a potential cause of RSM. Here, we perform liquid chromatography with mass spectrometry (LC-MS)-based metabolite profiling in human endometrial stromal cells (ESCs) and differentiated ESCs (DESCs) and find that accumulated α-ketoglutarate (αKG) derived from activated glutaminolysis contributes to maternal decidualization. Contrarily, ESCs obtained from patients with RSM show glutaminolysis blockade and aberrant decidualization. We further find that enhanced Gln-Glu-αKG flux decreases histone methylation and supports ATP production during decidualization. In vivo, feeding mice a Glu-free diet leads to a reduction of αKG, impaired decidualization, and an increase of fetal loss rate. Isotopic tracing approaches demonstrate Gln-dependent oxidative metabolism as a prevalent direction during decidualization. Our results demonstrate an essential prerequisite of Gln-Glu-αKG flux to regulate maternal decidualization, suggesting αKG supplementation as a putative strategy to rectify deficient decidualization in patients with RSM.

Antitussive, expectorant, and anti-inflammatory activities of four caffeoylquinic acids isolated from Tussilago farfara.

CONTEXT: The flower bud of Tussilago farfara L. (Compositae) (FTF) is one of the traditional Chinese medicinal herbs used to treat cough, phlegm, bronchitic, and asthmatic conditions. OBJECTIVE: The objective of this study is to isolate four caffeoylquinic acids from the ethyl acetate extract (EtE) of FTF and to evaluate their antitussive, expectorant, and anti-inflammatory activities. MATERIALS AND METHODS: The structures of compounds 1-4 isolated from EtE were determined by spectral analysis. Mice were orally treated with these compounds and their mixture (in a ratio of 5:28:41:26 as in EtE) at doses of 10 and 20 mg/kg once daily for 3 d. The antitussive and expectorant activities were evaluated separately with the ammonia liquor-induced model and the phenol red secretion model. The anti-inflammation activity was evaluated using leukocyte count in the bronchoalveolar lavage fluid after ammonia liquor-induced acute airway inflammation. RESULTS: The four compounds were identified as chlorogenic acid (1), 3,5-dicaffeoylquinic acid (2), 3,4-dicaffeoylquinic acid (3), and 4,5-dicaffeoylquinic acid (4). All compounds, especially compound 4 (58.0% inhibition in cough frequency), showed a significant antitussive effect. However, the mixture was the most effective to inhibit the cough frequency by 61.7%. All compounds also showed a significant expectorant effect, while compound 2 was the most potent to enhance the phenol red secretion by 35.7%. All compounds significantly alleviated inflammation, but compound 4 showed the strongest effect to inhibit the leukocytosis by 49.7%. DISCUSSION AND CONCLUSION: The caffeoylquinic acids and their mixture, exhibiting significant antitussive, expectorant, and anti-inflammatory effects, could be considered as the main effective ingredients of FTF, and they may act in a collective and synergistic way.

[Morphological and TLC identification on Tibetan medicine Asteris Flos].

Tibetan medicine Asteris Flos is the flowers of Aster souliei, A. flaccidus, and A. asteroides, with the function of clearing away heat and toxic matter, relieving cough, and removing phlegm. In order to control the quality of Asteris Flos, the morphological and chemical methods were established for identification of three origins. The morphological features of three species were described and photographed, and the microscopic characteristics of three drug powders were also described in detail and pictured. The results showed that three origins of Asteris Flos could be easily distinguished by their macro- and micro-morphologic features, and a key for distinguishing the three origins was given. Moreover, a TLC method, with apigenin-7-O-ß-D-glucurono pyranoside and chlorogenic acid as chemical references, was also established for the identification of three origins. The results showed that the TLC chromatograms of the flowers of A. souliei and A. flaccidus were very similar, but different from that of A. asteroides. The established macroscopic, powder microscopic and TLC methods for identification of three origins of AF were simple, accurate, and reproducible, and also effective and easy to operate.

Preventive effects of Ecliptae Herba extract and its component, ecliptasaponin A, on bleomycin-induced pulmonary fibrosis in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Ecliptae Herba, a nourishing traditional Chinese medicine, is also a folk medicine for the treatment of lung diseases. AIM OF THE STUDY: To investigate the anti-fibrosis effects and the underlying mechanism of the extract of Ecliptae Herba and its potential active components. MATERIALS AND METHODS: The resulting extract (EXT) was prepared from the 80% ethanol extract of Ecliptae Herba. After intratracheally administrated with bleomycin (BLM, 5mg/kg), mice were orally treated with EXT at 2.5, 1.25, 0.625 g/kg and eclipta saponin A (ESA) at 80 mg/kg once daily for 28 day. The bodyweight, survival rate, pathological changes of lung and levels of hydroxyproline (HYP) were used to evaluate the anti-fibrotic effects. The malonaldehyde (MDA), superoxidae dismutase (SOD) activity, and the protein expressions of matrix metalloproteinase (MMP)-2, 9, tissue inhibitor of metalloproteinase-1 (TIMP-1), cyclooxygenase-2 (COX-2), α-smooth muscle actin (α-SMA) and transforming growth factor-ß1 (TGF-ß1) in lung tissue were analyzed by kits or western blot. RESULTS: Compared with BLM group, EXT administration could significantly ameliorated the pathological changes of lung, decreased the HYP content, enhanced the SOD activity, and reduced the MDA content of lung tissues. In mechanism, EXT significantly alleviated the levels of COX-2, TGF-ß1, MMP-2 and α-SMA, as well as elevated the ratio value of MMP-9/TIMP-1. Additionally, the anti-fibrosis effects of ESA, a large amount of saponins isolated from Eclipta prostrata , was also evaluated by the BLM-induced model. The results showed that ESA could block BLM-induced histological changes of lung tissue and decrease the high levels of TGF-ß1 and α-SMA. CONCLUSIONS: Ecliptae Herba has protective effects against the pulmonary fibrosis induced by BLM via reducing the oxidative stress, lung tissue inflammation, and the subsequent epithelial-mesenchymal transition. The active chemical constituents may be involved with triterpenoid saponins, such as ESA.

Immunosuppressive C21 steroidal glycosides from the root of Cynanchum atratum.

Six new C21 steroidal glycosides (1-6) and one dideoxysaccharide (7), named atratcynosides A-F and atratcynose A, were isolated from the 80% ethanol extract of the root of Cynanchum atratum, together with three known compounds (8-10). The structures of the new compounds were determined on the basis of extensive spectral analyses and qualitative chemical methods. All compounds were subjected to detect the immunosuppressive activities by an in vitro model of concanavalin A-induced proliferation of T-lymphocytes from mice. Compounds 1-3 showed significant immunosuppressive activities in dose-dependent manners with the IC50 values from 3.3 to 7.0 µM. Moreover, the structure-activity relationship of the steroidal glycosides on the immunosuppression was analyzed.

Simultaneous separation and determination of phenolic acids, pentapeptides, and triterpenoid saponins in the root of Aster tataricus by high-performance liquid chromatography coupled with electrospray ionization quadrupole time-of-flight mass spectrometry.

We established a qualitative method to analyze the main chemical compositions of the root of Aster tataricus. Most of the peaks were separated on a C(18) column packed with 5.0 µm particles, and 28 compounds were identified, including 11 chlorogenic acids, ten astins/asterinins, and seven astersaponins, four of which were reported for the first time from A. tataricus. Furthermore, we developed a reliable method for the simultaneous quantification of 3-caffeoylquinic acid, 3,4-dicaffeoylquinic acid, 3,5-dicaffeoylquinic acid, astin A, astin B, astin C, astersaponin A, and astersaponin C, and the qualified separations were achieved only on a C18 column packed with 2.7 µm particles. The method was used to measure the concentrations of eight components in samples from two major producing areas in China, and the average contents in samples from Bozhou (Anhui) were higher than those in samples from Anguo (Hebei).

Astin B, a cyclic pentapeptide from Aster tataricus, induces apoptosis and autophagy in human hepatic L-02 cells.

Astins (including astin B) are a class of halogenated cyclic pentapeptides isolated from the medicinal herb of Aster tataricus. However, our previous works showed that the herbal medicine was hepatotoxic in vivo, and a toxicity-guided isolation method led to the identification of a cyclopeptide astin B. Astin B is structurally similar to cyclochlorotine, a well-known hepatotoxic mycotoxin. Thus, the aim of this study was to determine the potential cytotoxic effects and the underlying mechanism of astin B on human normal liver L-02 cells. We found that astin B has hepatotoxic effects in vitro and in vivo and that hepatic injury was primarily mediated by apoptosis in a mitochondria/caspase-dependent manner. Astin B provoked oxidative stress-associated inflammation in hepatocytes as evidenced by increased levels of reactive oxygen species (ROS), reduced contents of intracellular glutathione (GSH), and enhanced phosphorylation of c-Jun N-terminal kinase (JNK). Furthermore, the mitochondria-dependent apoptosis was evidenced by the depolarization of the mitochondrial membrane potential, the release of cytochrome c into cytosol, the increased ratio of Bax/Bcl-2, and the increased activities of caspases-9 and -3. Interestingly, astin B treatment also induces autophagy in L-02 cells, characterized by acidic-vesicle fluorescence, increased LC3-II and decreased p62 expression. Autophagy is a protective mechanism that is used to protect cells from apoptosis. The presence of autophagy is further supported by the increased cytotoxicity and the enhanced cleaved caspase-3 after co-treatment of cells with an autophagy inhibitor, also by increased LC3-II and decreased p62 after co-treatment with a caspase inhibitor. Taken together, astin B, most likely together with other members of astins, is the substance that is primarily responsible for the hepatotoxicity of A.tataricus.

[Chemical constituents from the aerial part of Sibiraea angustata].

OBJECTIVE: To study the chemical constituents from the aerial part of Sibiraea angustata. METHODS: The constituents were isolated by various chromatographic techniques (HP-20 macroporous absorption resin, Sephadex LH-20 gel, RP-MPLC and PHPLC)and their structures were determined on the basis of physicochemical properties and their spectroscopic data,as well as literatures. RESULTS: Eleven compounds were separated and identified as p-methoxycinnamic acid(I), protocatechuic aldehyde(II), quercetin(III), isorhamnetin(IV), quercetin 3-O-beta-D-galactopyranoside (V),9-0-[beta-D-glucopyranoside]-3,4,5-trimethoxy cinnamyl alcohol(VI), syringaresinol-4'-O-beta-D-monoglucoside(VII), ntin(VIII), sibiraic acid(IX), sibiscolacton(X), methyl ferulic acid(XI). CONCLUSION: Compounds I-XIII are isolated from the genus of Sibiraea for the first time.

[A new anthraquinone from the root of Lasianthus acuminatissimus].

To study the constituents from the chloroform extract of the roots of Lasianthus acuminatissimus Merr., various chromatographic techniques were used to separate and purify the constituents. The structure was established on the basis of ID, 2D NMR and HRMS spectroscopic analysis. A new compound was isolated and identified, which was 3, 8-dihydroxy-1-methoxy-2-methoxymethyl-9,10-anthraquinone (I). Compound I is a new anthraquinone, namely lasianthurin.

[Resource investigation and quality evaluation on wild Corydalis yanhusuo].

OBJECTIVE: To study the resource and the quality of wild Corydalis yanhusuo distributed in China. METHOD: Distribution was observed and samples of wild C. yanhusuo were collected and qualities were evaluated by determining six main alkaloids contained in the samples. RESULT: The main distribution of wild Corydalis yanhusuo was in the hills around middle and lower reaches of Yangtze River. Its distributive areas were continuously decreasing. The alkaloids contents in the samples varied among different populations. CONCLUSION: The alkaloids contents in wild populations of C. yanhusuo are diverse. The main kinds of alkaloids in some wild populations are higher than cultivated ones, which are valuable for breeding. The wild C. yanhusuo propagate well, however they are endangered due to environment problem, and should be protected.