BackgroundPulmonary 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.PurposeTo 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.MethodsA 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 β1-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.ResultsPCF-EtOH ameliorated bleomycin-induced pulmonary fibrosis through repressing inflammatory response and extracellular matrix deposition. Meanwhile, PCF-EtOH inhibited Wnt/β-catenin pathway through decreasing β-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/β-catenin pathway. Notably, physapubescin demonstrated a more potent antifibrotic effect than pirfenidone, a clinically approved antifibrotic drug, in the tested model.ConclusionWithanolides 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.

Background: Acute lung injury (ALI) is a high-fatality respiratory disease, and the development of new therapeutic agents is ongoing. Chuanxiong Rhizoma has been prescribed for the therapy of ALI in traditional Chinese medicine. Objective: The purpose of this article is to explore the effective ingredients, targets, and mechanisms in the treatment of Chuanxiong Rhizoma on ALI. Methods: The active ingredients of Chuanxiong Rhizoma were obtained in the TCMSP, and their potential targets and ALI targets were predicted in TCMSP, Swiss target prediction, and Genecards database. A PPI network was constructed in the string online platform, and KEGG and GO enrichment analysis were conducted in the metascape online platform. Phytochemical investigation of Chuanxiong Rhizoma was performed with chemical separation methods and structural elucidation techniques, and their pharmacological effects were evaluated using LPS-induced mouse ALI model and cell-based assays. Results: Network pharmacology analyses indicated that the treatment of Chuanxiong Rhizoma on ALI was associated with its anti-inflammatory effect. The key genes mainly include TNF, STAT3, EGFR, AKT1, BCL2, CASP3, and SRC, and the active compounds are its signature component, phthalides. Phytochemical investigation led to the elucidation of 23 phthalides, which demonstrated an anti-inflammatory effect in vitro and in vivo. Of which, chuanxiongdiolide A, levistilide A, butylphthalide, and senkyunolide A were indeed effective in mitigating ALI in an LPS-induced mouse model. Conclusions: The combined network pharmacological, phytochemical, and pharmacological investigations revealed that the therapeutical effect of Chuanxiong Rhizoma against ALI was associated with inhibition of the inflammatory response by phthalides.

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.

ETHNOPHARMACOLOGICAL RELEVANCE:In many famous formulas of traditional Chinese medicine (TCM), the rhizome of Ligusticum chuanxiong (L. chuanxiong) is commonly used as an ingredient for promoting blood circulation and resolving blood stasis to treat diabetic nephropathy. However, its material basis and mechanism of action are still needed to be explored.AIM OF THE STUDY:The aim of this work is to elucidate the potential effective parts (phthalides) of L. chuanxiong responsible for renal protection and to explore the possible mechanism of renal protection.MATERIALS AND METHODS:A method based on column chromatography of macroporous resin was established to enrich an effective part (LCE70), and the composition of LCE70 was identified by HPLC-UV and UPLC-MS/MS methods. Mice model was induced by streptozotocin (STZ) to evaluate the protective effect of LCE70 on diabetic nephropathy (DN). In vitro, the suppressive effect of LCE70 on oxidative damage, inflammation and its mechanism were tested using immunoblot analysis, ELISA, etc. Cellular thermal shift assay (CETSA) was adopted to verify the interaction between the phthalides and the key targets involved in renal injury.RESULTS:LCE70 displayed therapeutic potential against metabolic disorders, renal dysfunction, and fibrosis in a DN model induced by STZ in mice. Furthermore, it markedly reduced oxidative stress of the kidney in DN mice by activating Nrf2 pathway. Z-ligustilide, the main component of LCE70, reacted with Keap1, and thus promoted Nrf2 dissociating from Keap1 to activate Nrf2 pathway.CONCLUSIONS:LCE70 improved hyperglycemia-induced renal function by enhancing the Nrf2 activation, reducing collagen deposition, and alleviating inflammation and oxidative stress, which suggested its potential as a therapeutic agent for DN.

Liverworts provide valuable ecological services to improve the sustainability of agriculture, encompassing soil health maintenance and natural pest management. Some liverworts have potential applications in medicine and as food additives. Twenty-two novel diterpenoids (anajoerins A-V), of which anajoerins B-G are rearranged labdanes featuring an unprecedented 6/5 fused ring system, were isolated from the Chinese liverwort Anastrophyllum joergensenii Schiffn. The absolute configurations of all compounds were identified based on high-resolution electrospray ionization mass spectroscopy data, NMR spectra, and ECD calculations. Plausible biogenetic pathways for unprecedented rearranged labdanes were proposed. Seven diterpenoids exhibited anti-inflammatory activity by reducing nitric oxide production in LPS-stimulated RAW264.7 murine macrophages in a dose-dependent manner with IC50s between 9.71 and 56.56 μM. All tested compounds showed no cytotoxicity at the tested concentrations. Western blot analyses of NF-κB p65 downregulation showed that anajoerin L could inhibit the NF-κB signaling pathway. Furthermore, anajoerin L also suppressed the secretion of the ConA-induced proinflammatory cytokines IFN-γ, TNF-α, and IL-6.

Ethnopharmacological relevance: The flower buds of Tussilago farfara L. (Abbreviated as FTF) were widely used in traditional Chinese medicine (TCM) to treat respiratory diseases, including asthma, dry throat, great thirst, turbid saliva, stinky pus, and coughs caused by various causes. Aim of study: The aim of study is to explore the efficiency of FTF in vitro and in vivo for the treatment of lung inflammation, and to illustrate the possible mechanisms of FTF in treating inflammation-related respiratory diseases targeting NOD-like receptor 3 (NLRP3) inflammasome, nuclear factor erythroid 2-related factor 2 (Nrf2), and nuclear transcription factor-kappa B (NF-kappa B). Methods: Lung inflammation model in vivo was induced by exposure of mice to cigarette smoke (CS) for two weeks. The levels of superoxide dismutase (SOD), malondialdehyde (MDA), inflammatory factors, and histology in lung tissues were investigated in presence or absence of ethanol extract of the flower buds of T. farfara L. (FTF-EtOH). In the cell-based models, nitric oxide (NO) assay, flow cytometry assay, enzyme-linked immunosorbent assay (Elisa), and glutathione (GSH) assay were used to explore the anti-inflammatory and anti-oxidant effects of FTF-EtOH. Possible anti-inflammatory mechanisms of FTF targeting NLRP3 inflammasome, Nrf2, and NF-kappa B have been determined using western blot, quantitative real-time reverse transcriptase-polymerase chain reaction (qRT-PCR), immunofluorescence assay, nuclear and cytoplasmic extraction, and ubiqutination assay. Results: FTF-EtOH suppressed CS-induced overproduction of inflammatory factors [e.g., tumor necrosis factor-alpha (TNF-alpha) and interleukin-1 beta (IL-1 beta)], and upregulation of the content of intracellular MDA in the lung homogenate of mice. In cell-based models, FTF-EtOH reduced the lipopolysaccharide (LPS)-induced overproduction of inflammatory factors, and attenuated the CS extract-induced overgeneration of reactive oxygen species (ROS). Furthermore, FTF-EtOH up-regulated Nrf2 and its downstream genes through enhancing the stability of Nrf2 protein, and inhibited the activation of NF-kappa B and NLRP3 inflammasome, which have been confirmed by detecting the protein levels in the mouse model. Conclusions: FTF-EtOH effectively attenuated lung inflammation in vitro and in vivo. The protection of FTF-EtOH against inflammation was produced by activation of Nrf2 and inhibitions of NF-kappa B and NLRP3 inflammasome. These datas definitely support the ethnopharmacological use of FTF as an anti-inflammatory drug for treating respiratory diseases in TCM.

Two new physalins, 7α-hydroxy-5-deoxy-4-dehydrophysalin IX (1) and 5-deoxy-4-dehydrophysalin IX (2), together with six known compounds, luteolin (3), luteolin-7-O-glucoside (4), neoechinulin A (5), 3-(4-hydroxy-3-methoxyphenyl)-N-(4-methylphenyl)-2-propenamide (6), physalin D (7) and blumenol A (8) were isolated from Physalis alkekengi L. var. franchetii (Mast.) Makino. Their structures were elucidated by NMR spectroscopic analysis, HR-ESI-MS, X-ray crystallographic data analysis and comparison with the known compounds. Among them, compounds 5 and 6 were isolated from the genus Physalis for the first time. Compound 1 exhibited weak NAD(P)H: quinone reductase (QR) inducing activity.

The rhizome of Ligusticum chuanxiong Hort. has been widely used for the therapy of diabetic nephropathy (DN) in traditional Chinese medicine (TCM). The nuclear transcription factor erythroid 2‐related factor (Nrf2) is a potential target for treating DN. The purpose of this research was to study the chemical constituents from the rhizome of L. chuanxiong, evaluate their Nrf2 inducing activity, and find the molecules with potential therapeutic effect against DN. In this study, two new phthalides (1–2) along with twenty‐seven known constituents were obtained from the rhizome of L. chuanxiong. Their structures were elucidated through various spectroscopic methods. Twelve constituents, including eight phthalides (2, 5, 6,10–13, 14) and four other compounds (17, 18, 20,28), stimulated NAD(P)H: quinone reductase (QR) activity, suggesting that these bioactive constituents were potential Nrf2 activators. Among the isolated compounds, phthalide levistolide A (LA, 14) upregulated the protein levels of Nrf2, NQO1, and γ‐GCS in a dose‐dependent manner. Our results implied that the clinical application of the rhizome of L. chuanxiong as an anti‐DN drug in TCM might be attributed to the Nrf2 inducing effect of phthalides. Thus, phthalides is a group of promising leading molecules for discovering anti‐DN agents.

Kinsenoside is the major bioactive component from herbal medicine with a broad range of pharmacological functions. Goodyeroside A, an epimer of kinsenoside, remains less explored. In this report we chemically synthesized kinsenoside, goodyeroside A and their analogues with glycan variation, chirality inversion at chiral center(s), and bioisosteric replacement of lactone with lactam. Among these compounds, goodyeroside A and its mannosyl counterpart demonstrated superior anti-inflammatory efficacy. Furthermore, goodyeroside A was found to suppresses inflammatory through inhibiting NF-κB signal pathway, effectively. Structure-activity relationship is also explored for further development of more promising kinsenoside analogues as drug candidates.

Acidic compounds were enriched from a water decoction of Portulaca oleracea using 717 anion exchange resin column chromatography. A total of 22 compounds including 9 catecholamine derivatives, of which six were rare sulfonic acid derivatives, and 9 nitro derivatives, were further isolated through various column chromatographic methods, and their structures were elucidated by interpreting their spectroscopic data and ECD calculations. Among them, 16 compounds were isolated from P. oleracea for the first time, 8 of which were undescribed compounds and four compounds were natural products. Pharmacological screening indicated that cis-3-(3-nitro-4-hydroxyphenyl)-methyl acrylate exhibited anti-inflammatory activity, measured as inhibition of nitric oxide production in LPS-stimulated RAW264.7 macrophage cells, with an EC50 value of 18.0 μM, The compounds showed only weak anti-microbial activity with (2R)-(+)-2-chloro-3-(3-nitro-4-hydroxyphenyl)-propionic acid methyl ester inhibiting Candida albicans with a MIC of 256 μg/mL, and 3-methoxy-4,5-dinitrophenol inhibiting Shigella sonnei with a MIC of 512 μg/mL.

Environmental toxicant- and oxidant-induced [e.g., cigarette smoke (CS)] respiratory oxidative stress and inflammatory response play a vital role in the onset and progression of COPD. The nuclear factor erythroid 2-related factor 2 (Nrf2) represents an important mechanism for regulating intracellular oxidative stress and inflammatory response and is a promising target for developing agents against COPD. Herein, a bioactivity-guided purification of goldenberry (whole fruits of Physalis peruviana L.) led to the isolation of a novel and potent Nrf2 activator 4β-hydroxywithanolide E (4β-HWE). Our study indicated that (i) 4β-HWE activated the Nrf2-mediated defensive response through interrupting Nrf2-Keap1 protein-protein interaction (PPI) via modification of Cys151 and Cys288 cysteine residues in Keap1 and accordingly suppressing the ubiquitination of Nrf2. (ii) 4β-HWE enhanced intracellular antioxidant capacity and inhibited oxidative stress in normal human lung epithelial Beas-2B cells and wild-type AB zebrafish. (iii) 4β-HWE blocked LPS-stimulated inflammatory response and inhibited LPS-stimulated NF-κB activation in RAW 264.7 murine macrophages. (iv) 4β-HWE effectively suppressed oxidative stress and inflammatory response in a CS-induced mice model of pulmonary injury. Collectively, these results display the feasibility of using 4β-HWE to prevent or alleviate the pathological progression of COPD and suggest that 4β-HWE is a candidate or a leading molecule against COPD.

Physalis Calyx seu Fructus, a traditional Chinese medicine consisting of the calyxes and fruits of Physalis alkekengi var. franchetii , has been used as therapy for inflammation-related respiratory diseases such as excessive phlegm, cough, sore throat, and pharyngitis for a long history in China. The aim of the present study was to investigate the chemical constituents of Physalis Calyx seu Fructus and identify the bioactive constituents responsible for its traditional application as therapy for inflammation-related diseases. In the present study, one new phenylpropanoid ( 1 ), two new steroids ( 17 and 18 ), together with 55 known constituents have been purified from the EtOH extract of Physalis Calyx seu Fructus. Among them, seven and twelve known constituents were isolated for the first time from Physalis Calyx seu Fructus and the genus Physalis , respectively. Fourteen constituents, including steroids [physalins ( 5 - 9, 12 - 14 , and 15 ) and ergostane ( 21 )], a sesquiterpenoid ( 35 ), alkaloids ( 36 and 37 ), and a flavonoid ( 44 ), showed inhibitory effects against oxidative stress. Ten constituents, including steroids ( 5, 6, 8, 13 , and 15 ), sesquiterpenoids ( 34 and 35 ), alkaloids ( 37 and 41 ), and a flavonoid ( 43 ), were found be potential anti-inflammatory constituents of this medicinal plant. The inhibition of oxidative stress and inflammatory response may be related to the regulation of Nrf2 and nuclear factor- kappa B pathways. The ethnomedical use of Physalis Calyx seu Fructus as a treatment for respiratory diseases might be attributed to the combined inhibitory effects of steroids, alkaloids, sesquiterpenoids, and flavonoids against oxidative stress and inflammatory response.

Seven undescribed polyketides javanicols A-E, 5-epi-citreoviridin and 5-epi-isocitreoviridin, together with five known compounds, were isolated from the endolichenic fungus Eupenicillium javanicum. The structures of these polyketides were determined by means of extensive spectroscopic analyses, electronic circular dichroism (ECD) calculations and gauge-independent atomic orbital (GIAO) NMR shift calculations. These compounds were evaluated for potential anti-inflammatory activity against LPS-activated RAW 264.7 cells. Javanicol E and (+)-terrein displayed moderate inhibitory effects on NO production, with IC50 values of 17.00 and 13.46 mu M, respectively.

Dracomolphesin A–E (1–5), five 3,4-seco-phenylpropanoids featuring an aromatic ring opened framework, were isolated from the aerial parts of Dracocephalum moldavica. The structures with absolute configurations were determined by spectroscopic methods coupled with Mosher method. Notably, these compounds represented an example of aromatic ring cleavage products of phenyl-propanoids. The possible biosynthetic pathway of these compounds was proposed. Compounds 1, 2, 4 and 5 were demonstrated to be Nrf2 pathway activators.