Ethanol Extract of Limonium bicolor Improves Dextran Sulfate Sodium-Induced Ulcerative Colitis by Alleviating Inflammation and Restoring Gut Microbiota Dysbiosis in Mice.

Ulcerative colitis (UC) is a kind of inflammatory bowel condition characterized by inflammation within the mucous membrane, rectal bleeding, diarrhea, and pain experienced in the abdominal region. Existing medications for UC have limited treatment efficacy and primarily focus on symptom relief. Limonium bicolor (LB), an aquatic traditional Chinese medicine (TCM), exerts multi-targeted therapeutic effects with few side effects and is used to treat anemia and hemostasis. Nevertheless, the impact of LB on UC and its mechanism of action remain unclear. Therefore, the objective of this study was to investigate the anti-inflammatory effects and mechanism of action of ethanol extract of LB (LBE) in lipopolysaccharide-induced RAW 264.7 macrophages and dextran sulfate sodium (DSS)-induced UC. The results showed that LBE suppressed the secretion of cytokines in LPS-stimulated RAW 264.7 cells in a dose-dependent manner. LBE had protective effects against DSS-induced colitis in mice, decreased the disease activity index (DAI) score, alleviated symptoms, increased colon length, and improved histological characteristics, thus having protective effects against DSS-induced colitis in mice. In addition, it reversed disturbances in the abundance of proteobacteria and probiotics such as Lactobacillus and Blautia in mice with DSS-induced UC. Based on the results of network pharmacology analysis, we identified four main compounds in LBE that are associated with five inflammatory genes (Ptgs2, Plg, Ppar-γ, F2, and Gpr35). These results improve comprehension of the biological activity and functionality of LB and may facilitate the development of LB-based compounds for the treatment of UC.

A polysaccharide from Allium tenuissimum L. flowers relieves ulcerative colitis by regulating the inflammatory signaling pathway and gut microbiota.

A polysaccharide fraction named ATFP was isolated and purified from Allium tenuissimum L. flowers. Its primary structure and therapeutic effects on mice with acute ulcerative colitis were investigated in the present study. The results showed that the molecular weight of ATFP was determined to be 1.56 × 106 Da without nucleic acids and protein. Moreover, ATFP was a pyranose type acidic polysaccharide containing α and ß type glycosidic bonds and consisted of Ara, Gal, Glc, Xyl, GlcA and Glca with molar percentages of 14.55 : 49.46 : 7.28 : 23.23 : 2.49 : 3.01. Microscopic observation revealed that ATFP had a smooth lamellar structure with pores and multiple molecular chains were intertwined. In animal experiments of dextran sodium sulfate-induced acute ulcerative colitis, the results indicated that ATFP significantly improved the symptom of weight loss, decreased the disease activity index and alleviated pathological damage. The anti-inflammatory mechanism of ATFP might be related to the inhibition of the TLR4/MyD88/NF-κB signaling pathway to regulate the level of inflammatory cytokines. In particular, ATFP also played an important role in regulating the structure of gut microbiota, which was specifically reflected in promoting the abundance of short-chain fatty acid producing bacteria. Overall, ATFP showed a significant mitigating effect against ulcerative colitis in mice, and it is expected to prove its value efficiently in the field of functional food.

Ramulus Cinnamomi essential oil exerts an anti-inflammatory effect on RAW264.7 cells through N-acylethanolamine acid amidase inhibition.

ETHNOPHARMACOLOGICAL RELEVANCE: Ramulus Cinnamomi, the dried twig of Cinnamomum cassia (L.) J.Presl., is a traditional Chinese medicine (TCM) with anti-inflammatory effects. The medicinal functions of Ramulus Cinnamomi essential oil (RCEO) have been confirmed, although the potential mechanisms by which RCEO exerts its anti-inflammatory effects have not been fully elucidated. AIM OF THE STUDY: To investigate whether N-acylethanolamine acid amidase (NAAA) mediates the anti-inflammatory effects of RCEO. MATERIALS AND METHODS: RCEO was extracted by steam distillation of Ramulus Cinnamomi, and NAAA activity was detected using HEK293 cells overexpressing NAAA. N-Palmitoylethanolamide (PEA) and N-oleoylethanolamide (OEA), both of which are NAAA endogenous substrates, were detected by liquid chromatography with tandem mass spectrometry (HPLC-MS/MS). The anti-inflammatory effects of RCEO were analyzed in lipopolysaccharide (LPS)-stimulated RAW264.7 cells, and the cell viability was measured with a Cell Counting Kit-8 (CCK-8) kit. The nitric oxide (NO) in the cell supernatant was measured using the Griess method. The level of tumor necrosis factor-α (TNF-α) in the RAW264.7 cell supernatant was determined using an enzyme-linked immunosorbent assay (ELISA) kit. The chemical composition of RCEO was assessed by gas chromatography-mass spectroscopy (GC-MS). The molecular docking study for (E)-cinnamaldehyde and NAAA was performed by using Discovery Studio 2019 software (DS2019). RESULTS: We established a cell model for evaluating NAAA activity, and we found that RCEO inhibited the NAAA activity with an IC50 of 5.64 ± 0.62 µg/mL. RCEO significantly elevated PEA and OEA levels in NAAA-overexpressing HEK293 cells, suggesting that RCEO might prevent the degradation of cellular PEA and OEA by inhibiting the NAAA activity in NAAA-overexpressing HEK293 cells. In addition, RCEO also decreased NO and TNF-α cytokines in lipopolysaccharide (LPS)-stimulated macrophages. Interestingly, the GC-MS assay revealed that more than 93 components were identified in RCEO, of which (E)-cinnamaldehyde accounted for 64.88%. Further experiments showed that (E)-cinnamaldehyde and O-methoxycinnamaldehyde inhibited NAAA activity with an IC50 of 3.21 ± 0.03 and 9.62 ± 0.30 µg/mL, respectively, which may represent key components of RCEO that inhibit NAAA activity. Meanwhile, docking assays revealed that (E)-cinnamaldehyde occupies the catalytic cavity of NAAA and engages in a hydrogen bond interaction with the TRP181 and hydrophobic-related interactions with LEU152 of human NAAA. CONCLUSIONS: RCEO showed anti-inflammatory effects by inhibiting NAAA activity and elevating cellular PEA and OEA levels in NAAA-overexpressing HEK293 cells. (E)-cinnamaldehyde and O-methoxycinnamaldehyde, two components in RCEO, were identified as the main contributors of the anti-inflammatory effects of RCEO by modulating cellular PEA levels through NAAA inhibition.

Medicinal Formula Huazhi-Rougan Attenuates Non-Alcoholic Steatohepatitis Through Enhancing Fecal Bile Acid Excretion in Mice.

Huazhi-Rougan (HZRG) formula is a Traditional Chinese medicine prescription, and has been widely used to treat non-alcoholic fatty liver disease (NAFLD) and its progressive form non-alcoholic steatohepatitis (NASH). However, the anti-NASH effects and the underlying mechanisms of HZRG have not yet been characterized. Here we showed that 4-week HZRG treatment alleviated methionine-choline-deficiency (MCD) diet-induced NASH in C57BL/6J mice, as evidenced by the improvement of hepatic steatosis and inflammation, as well as the decrease of serum levels of alanine and aspartate transaminases. Fecal 16S rDNA sequencing indicated that HZRG reduced the enrichment of pathogenic bacteria and increased the abundance of bacteria gena that are involved in bile acid (BA) conversation. The alteration of fecal and serum BA profile suggested that HZRG enhanced fecal BA excretion, and reduced the reabsorption of toxic secondary BA species (LCA, DCA, HCA). We further analyzed the BA receptors and transporters, and found that HZRG inhibited the expression of ileal bile acid transporter, and organic solute transporter subunit ß, and increased the expression of intestinal tight junction proteins (ZO-1, Occludin, Claudin-2). The modulation of gut dysbiosis and BA profile, as well as the improvement of the intestinal environment, may contribute to the decrease of the p-65 subunit of NF-κB phosphorylation, liver F4/80 positive macrophages, inflammatory cytokine IL-1ß and TNF-α expression. In conclusion, HZRG treatment enhances fecal BA excretion via inhibiting BA transporters, modulates BA profiles, gut dysbiosis as well as the intestinal environment, thus contributing to the beneficial effect of HZRG on NASH mice.

Jiangzhi granule attenuates non-alcoholic steatohepatitis through modulating bile acid in mice fed high-fat vitamin D deficiency diet.

Vitamin D deficiency is a common phenomenon in non-alcoholic fatty liver disease (NAFLD) and the progressive non-alcoholic steatohepatitis (NASH). Jiangzhi Granule (JZG) formula is a Traditional Chinese medicine prescription, and has been found effective against NAFLD/NASH. Here we showed that vitamin D deficiency could accelerate NASH development, and reduce vitamin D receptor (VDR) expression. JZG treatment alleviated high-fat vitamin D deficient (HF-VDD) diet-induced NASH in C57BL/6 J mice, and up-regulated both the liver and intestinal VDR expression independent of 1,25-dihydroxy-vitamin D3 level. We analyzed the fecal BA profile using liquid chromatography coupled with triple quadrupole mass spectrometry (UPLC-TQMS) -based metabolomics, and found that JZG modulated fecal BA profile, predominantly increased the ratio of secondary BA species, as well as the expression of tight junction proteins Zona occludens 1(ZO-1) and occludin in the colon. In vitro experiment further confirmed the representative secondary BA species lithocholic acid (LCA) and keto-LCA upregulated the expression of and ZO-1 through VDR in LPS-stressed Caco-2 cells. Our results identified the endogenous VDR activation by JZG through modulating BA species in vitamin D deficiency-related NASH mice, thus providing evidence for the clinical application of JZG in treating NASH.

Diet supplementation with iron augments brain oxidative stress status in a rat model of psychological stress.

OBJECTIVE: We investigated the influence of iron supplementation on brain oxidative stress and antioxidase activity in psychologically stressed rats. METHODS: Rats were maintained on diets with different iron doses for 1 wk, and all other constituents of the diet were equated exactly according to the AIN-93-G diet. At the end of the experimental period, rats were sacrificed and brains were collected. To evaluate the effect of iron consumption, serum iron, apparent iron absorption, levels of iron concentration, lipid peroxidation, reduced glutathione, and superoxide dismutase activities of brains were measured. RESULTS: Iron overload significantly elevated the level of iron content and malonaldehyde in rat brain, especially in the psychologically stressed group. Apparent iron absorption was decreased by increased iron supplementation in rats treated with psychological stress more than in control rats. Similarly, iron overload decreased superoxide dismutase activity and apparent iron absorption more significantly in psychologically stressed rats than in controls. Reduced glutathione level varied with diet, increasing in rats on a moderately high-iron diet but decreasing in rats on a extremely high-level iron diet. CONCLUSION: These results demonstrated that iron overload augments brain oxidative stress status and aggravates the decrease of apparent iron absorption in a rat model of psychological stress.