Physalis Calyx seu Fructus inhibited pulmonary fibrosis through regulating Wnt/ß-catenin signaling pathway.

BACKGROUND: Pulmonary fibrosis is a chronic and advancing interstitial lung disease, and there is an urgent need for novel agents for its therapy. Physalis Calyx seu Fructus (PCF) has been utilized in traditional Chinese medicine to treat respiratory disorders with a long history, however, the therapeutic effect and mechanism of PCF against pulmonary fibrosis are still unclear. PURPOSE: To assess therapeutic efficacy and underlying mechanism of 75 % ethanol extract of PCF (PCF-EtOH) against pulmonary fibrosis, as well as to discover active constituents in PCF. METHODS: A bleomycin-stimulated mice model was established to assess potential therapy of PCF-EtOH against pulmonary fibrosis in vivo. A lipopolysaccharide-induced inflammatory model in RAW 264.7 cells and a transforming growth factor ß1-induced fibrosis model in MRC-5 cells were established to assess potential therapy and mechanisms of purified constituents in PCF-EtOH. UPLC-MS/MS analysis was adopted to ascertain the constituents of PCF-EtOH. Network pharmacology was employed to forecast targets of PCF against pulmonary fibrosis. RESULTS: PCF-EtOH ameliorated bleomycin-induced pulmonary fibrosis through repressing inflammatory response and extracellular matrix deposition. Meanwhile, PCF-EtOH inhibited Wnt/ß-catenin pathway through decreasing ß-catenin nuclear accumulation and promoting phosphorylation. Furthermore, withanolides and flavonoids were presumed to be main active compounds of PCF against pulmonary fibrosis based on the network pharmacology. Importantly, we found an extensive presence of withanolides in PCF-EtOH. Physapubescin, a typical withanolide in PCF-EtOH, inhibited the inflammatory response, extracellular matrix deposition, and Wnt/ß-catenin pathway. Notably, physapubescin demonstrated a more potent antifibrotic effect than pirfenidone, a clinically approved antifibrotic drug, in the tested model. CONCLUSION: Withanolides and flavonoids are responsible for the inhibitory effect of PCF-EtOH against pulmonary fibrosis. Withanolides may represent a class of promising therapeutic agents against pulmonary fibrosis, and an in-depth exploration is warranted to validate this proposition.

Integrated network pharmacology and pharmacological investigations to explore the potential mechanism of Ding-Chuan-Tang against chronic obstructive pulmonary disease.

ETHNOPHARMACOLOGICAL RELEVANCE: Ding-Chuan-Tang (Abbreviated as DCT) is frequently prescribed for treatment of respiratory diseases, including chronic obstructive pulmonary disease (COPD), which is characterized by coughing, wheezing, and chest tightness in traditional Chinese medicine (TCM). However, the potential mechanism of DCT has not been investigated. AIM OF STUDY: The aim of the study is to explore the efficiency of DCT in the treatment of COPD in vivo and in vitro, and to illustrate the possible mechanism against COPD. METHODS: COPD model was induced by exposure of mice to cigarette smoke (CS) for 16 weeks. Enzyme-linked immunosorbent assay (ELISA), immunofluorescence assay, Western blot, etc., were used to explore the efficiency and mechanisms of DCT. Network pharmacology analysis, including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis, etc., was performed to explore the potential targets in the treatment of DCT on COPD. RESULTS: DCT significantly alleviated pulmonary pathological changes in mouse COPD model, and inhibited inflammatory response induced by CS and LPS in vivo and in vitro. Network pharmacology analysis suggested that DCT alleviated COPD via inhibiting inflammation by regulating PI3K-AKT pathway. In cell-based models, DCT suppressed the phosphorylation of PI3K and AKT, which further regulated its downstream targets Nrf2 and NF-κB, and inhibited inflammatory response. CONCLUSIONS: DCT effectively attenuated COPD in the mouse model induced by CS. The therapeutic mechanism of DCT against COPD was closely associated with the regulation of PI3K-AKT pathway and its downstream transcription factors, Nrf2 and NF-κB.

The toxic natural product tutin causes epileptic seizures in mice by activating calcineurin.

Tutin, an established toxic natural product that causes epilepsy in rodents, is often used as a tool to develop animal model of acute epileptic seizures. However, the molecular target and toxic mechanism of tutin were unclear. In this study, for the first time, we conducted experiments to clarify the targets in tutin-induced epilepsy using thermal proteome profiling. Our studies showed that calcineurin (CN) was a target of tutin, and that tutin activated CN, leading to seizures. Binding site studies further established that tutin bound within the active site of CN catalytic subunit. CN inhibitor and calcineurin A (CNA) knockdown experiments in vivo proved that tutin induced epilepsy by activating CN, and produced obvious nerve damage. Together, these findings revealed that tutin caused epileptic seizures by activating CN. Moreover, further mechanism studies found that N-methyl-D-aspartate (NMDA) receptors, gamma-aminobutyric acid (GABA) receptors and voltage- and Ca2+- activated K+ (BK) channels might be involved in related signaling pathways. Our study fully explains the convulsive mechanism of tutin, which provides new ideas for epilepsy treatment and drug development.

Flavonoid alkaloids from Scutellaria moniliorrhiza with anti-inflammatory activities and inhibitory activities against aldose reductase.

Four undescribed flavonoid alkaloids, as two pairs of enantiomers, were initially isolated as a racemate from the whole plant of Scutellaria moniliorrhiza. By means of chiral HPLC, four isomers, named scumonilines A-D, were successfully separated, and their chemical structures including absolute configurations were established by mass as well as NMR spectroscopy and CD technique. In vitro, four flavonoid alkaloids showed anti-inflammatory activities, with IC50 values against the release of ß-glucuronidase from polymorphonuclear leukocytes of rats being in the range 5.16-5.85 µΜ. Moreover, four compounds were evaluated for their inhibitory activities against aldose reductase, and gave IC50 values in the range 2.29-3.03 µΜ.

New Flavonoid Glucuronate Esters with Anti-inflammatory Activities from Scutellaria regeliana.

Two new flavonoid glucuronate esters, named scuregeliosides A and B (1 and 2), as well as three known ones, chrysin-7-O-ß-d-glucuronic acid methyl ester (3), 5,7,4'-trihydroxyflavone-8-O-ß-d-glucuronic acid methyl ester (4) and apigenin-7-O-ß-d-glucuronic acid ethyl ester (5), were isolated from the ethanolic extract of the whole plant of Scutellaria regeliana. Their chemical structures were elucidated on the basis of comprehensive spectroscopic analyses. Five compounds were screened for anti-inflammatory activity in vitro. As the results, the inhibition rates of release of ß-glucuronidase from rat polymorphonuclear leukocytes were in the range of 42.2 - 47.1% at a concentration of 10 µm.

[Studies on flavonoids from Scutellaria moniliorrhiza].

By means of preparative HPTLC and column chromatography over silica gel and Sephadex LH-20, nineteen flavonoids were isolated and purified from the whole plants of Scutellaria moniliorrhiza. Based on the physico-chemical properties and spectral data, their structures were identified as： apigenin (1), luteolin (2), wogonin (3), oroxylin A (4), 6-methoxynaringein (5), 5,7,4'-trihydroxy-6,8-dimethoxyflavone (6), 5,7,8-trimethoxyflavone (7), 3,5,6,7-tetramethoxyflavone (8), 7-hydroxy-4',5,6,8-tetramethoxyflavone (9), 5,7,2'-trihydroxy-6-methoxyflavanone (10), 5,7,4'-trihydroxy-6-methoxyflavone (11), 5,7-dihydroxy-6,8-dimethoxy -flavone (12), 5,2',6'-trihydroxy-7,8-dimethoxyflavone (13), 5,7,2'-trihydroxy-8-methoxyflavone (14), 5,2'-dihydroxy-7,8-dimethoxyflavanone (15), 2'-hydroxy-5,7,8-trimethoxyflavone (16), 5-hydroxy-7,8-dimethoxyflavone (17), 5,2'-dihydroxy-7,8-dimethoxyflavone (18), and 5-hydroxy-6,7,8-trimethoxyflavone (19). For the first time, compounds 1-19 were isolated from S. moniliorrhiza, and compounds 6, 8, 9, 12, 19 were isolated from the Scutellaria genus.

New cytotoxic neo-clerodane diterpenoids from Scutellaria strigillosa.

Three new neo-clerodane diterpenoids, named scutestrigillosins A-C (1-3), were isolated from the whole plant of Scutellaria strigillosa. Their chemical structures including absolute configurations were established on the basis of detailed physical data analyses. In vitro, the isolated three new compounds exhibited significant cytotoxic activities against four tumor cell lines (HONE-1, P-388, MCF7 and HT29), and gave IC50 values in the range 3.5-7.7µM.

New neo-clerodane diterpenoids from Scutellaria strigillosa with cytotoxic activities.

Two new neo-clerodane diterpenoids, named scutestrigillosins D and E (1 and 2), were isolated from the whole plant Scutellaria strigillosa. Their structures were established on the basis of detailed physical data analyses. In vitro, two new compounds exhibited cytotoxic activities against four tumor cell lines (HONE-1, P-388, MCF7, and HT29), and gave IC50 values in the range of 3.4-8.9 µΜ.

[neo-Clerodane diterpenoids from Scutellaria galericulata].

By means of preparative HPTLC and column chromatography over silica gel and Sephadex LH-20, fourteen diterpenoids were isolated and purified from the whole plants of Scutellaria galericulata. Based on the physico-chemical properties and spectral data, their structures were elucidated and identified as：scutebarbatine D(1), scutolide A(2), scutolide K(3), scutebata J(4), scutebata I(5), 6-O-acetylscutehenanine A(6), barbatin C(7), scutolide E(8), barbatine C(9), scutebarbatine Y(10), scutebarbatine B(11), scutestrigillosin A(12), scutebata O(13), scutolide B(14). Compounds 1-14 were isolated from S. galericulata for the first time.