Anti-inflammatory effect of flavonoids from chestnut flowers in lipopolysaccharide-stimulated RAW 264.7 macrophages and acute lung injury in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Chestnut flowers were one of the by-products during chestnut industrial processing. Chestnut (Castanea mollissima Blume) flower is rich in flavonoids and has been used as a traditional medicine to treat a variety of diseases including respiratory disorders for a long history. AIM OF THE STUDY: The present study aims to investigate the potential anti-inflammatory effect of flavonoids from chestnut flower (FCF) in lipopolysaccharide (LPS)-treated RAW 264.7 cells and stimulated acute lung injury (ALI) in mice. MATERIALS AND METHODS: HPLC-ESI-MS/MS was applied to identify flavonoids from Chestnut flower. The ROS content in cells and lung tissue was measured by flow cytometry. The malondialdehyde (MDA) content, superoxide dismutase (SOD) activity and glutathione (GSH) content in cells and bronchoalveolar lavage fluid (BALF) was analyzed by photometry. Furthermore, the level of pro-inflammatory factors was analyzed by ELISA, and the expression of inflammatory gene mRNA by fluorescence quantitative PCR. H&E staining was used to evaluate the degree of lung tissue injury in mice. MPO activity was used to measure the degree of neutrophil infiltration. Total protein content was detected by BCA method. RESULTS: A total of forty-nine flavonoids compounds were tentatively identified in FCF by mass spectrometry analysis. The results of cell experiment suggested that FCF could alleviate oxidative injury via increasing SOD activity and GSH content, as well as inhibiting the production of intracellular ROS and MDA. FCF exerted its protective effect by suppressing the expression of both inducible nitric oxide synthase (iNOS) and cycooxygenase 2 (COX-2) to inhibit the synthesis of pro-inflammatory factors and cytokines, including NO, PGE2, TNF-α, IL-6 and IL-1ß. Besides, FCF treatment could alleviate the thickening of alveolar wall and pulmonary congestion in LPS-treated ALI mice, and significantly inhibit the activity of myeloperoxidas (MPO) and the expression of cytokines in BALF. CONCLUSIONS: FCF could ameliorate inflammation and oxidative stress in LPS-treated inflammation, resulting in an overall improvement in both macroscopic and histological parameters.

Compositional characterization of Pyrus ussuriensis Maxim and their antioxidant activities and induction of apoptosis in Bel-7402 cell.

In this work, a comparison study on active sites (the total phenolic, total flavonoids, and total triterpenes contents) and antioxidant activities (DPPH, ABTS, and FRAP assays) of different fractions from Pyrus ussuriensis Maxim was evaluated. Moreover, the inhibition capability on human hepatocarcinoma cells Bel-7402 cells and the mechanism was also discussed. Results showed that the ethyl acetate fraction significantly scavenged DPPH and ABTS+ radicals, exhibited ferric ion reducing antioxidant, and inhibited Bel-7402 cells proliferation. In addition, oleanolic acid was the dominant compound act on the Bel-7402 cells in the extract and it induced apoptosis by the caspase pathway and induced cell cycle arrest at the G0/G1 phase by inhibiting the cyclin D1/CDK4 pathway. The extracts of P. ussuriensis Maxim were confirmed to have anti-oxidative and antiproliferative effects against Bel-7402 cell in vitro. PRACTICAL APPLICATIONS: Fruits and vegetables which contain high levels of antioxidants can help to reduce the risk of chronic diseases such as cancer. Pyrus ussuriensis Maxim is a kind of edible and medical fruit with multiple bioactivities, whereas the capability to anti-lung cancer activity has not been investigated. The extracts of P. ussuriensis Maxim were revealed to have anti-oxidative and antiproliferative effects against Bel-7402 cell in vitro. Accordingly, it is the first time to verify that oleanolic acid was the main chemical components of P. ussuriensis with antitumor potential.