Therapeutic effect of Patrinia villosa on TNBS-induced ulcerative colitis via metabolism, vitamin D receptor and NF-κB signaling pathways.

ETHNOPHARMACOLOGICAL RELEVANCE: Ulcerative colitis (UC) is a chronic inflammatory disease affecting the colon. Patrinia villosa Juss. (P.V) is an important traditional Chinese medicine widely used for more than 2000 years from ShenNongBenCaoJing, a famous ancient Chinese medicinal literary. P.V is often used in the treatment of UC, but the pathogenesis is not clear. AIM OF THE STUDY: This study was designed to analysis the metabolic pathways and relevant mechanisms of P.V on UC rats induced by TNBS. MATERIALS AND METHODS: The rat model of UC was established by 2,4,6-trinitrobenzene sulfonic acid (TNBS)/ethanol method. Three doses of P.V (21 g/kg, 43 g/kg, 64 g/kg) were administrated for 14 days. Disease activity index (DAI) scoring system and H&E staining were used to evaluate the efficacy. A method for simultaneous detection of 96 endogenous metabolic components was established by UPLC-MS. The method was used to detect the metabolites in serum and liver of rats with UC induced by TNBS. PLS-DA and Metaboanalyst were used to analyze the main metabolic pathways involved in the treatment of UC. The contents of IL-1ß, TNF-α and IL-6 in the colonic homogenate of rats were detected by enzyme-linked immunosorbent assay (ELISA). The expression levels of VDR, NF-κB, p-NF-κB, NLRP3 and caspase-1 in colon tissues of rats were detected by the method of Western blot. RESULTS: DAI scoring system and H&E staining indicated that P.V have the obvious therapeutic effect on UC induced by TNBS as a dosage-dependent manner. 36 potential biomarkers in serum and 26 potential biomarkers in liver were found in positive and negative ion mode of UPLC-MS, which significantly affected Glycine, serine and threonine metabolism, Glycerophospholipid metabolism, Purine metabolism, Histidine metabolism, Alanine, aspartate, and glutamate metabolism, Arginine and proline metabolism in serum, and significantly affected Purine metabolism, Lycine, serine and threonine metabolism, Glutathione metabolism, Glyoxylate and dicarboxylate metabolism in the liver. The contents of pro-inflammatory cytokines and receptors related to NF-κB signaling axis of model group were significantly higher than those of the control group, compared with the model group, their contents of the P.V group were significantly decreased (p < 0.01). Compared with the model group, the expression of NF-κB, p-NF-κB, NLRP3 and caspase-1 in colon tissues of the rats in P.V group were significantly decreased (p < 0.01). The expression of VDR in model group were significantly reduced compared to that in the control group, compared with the model group, the expression of VDR in P.V group were significantly increased (p < 0.01). CONCLUSION: P.V has an obvious therapeutic effect on UC induced by TNBS by regulating the energy metabolism, amino acid metabolism, purine metabolism, bile acid metabolism and lipid metabolism. P.V exerts anti-inflammatory effect by impacting bile acid levels, activating VDR, and inhibiting the overactivation of NF-κB signaling pathways.

Inhibitory effect of main phenolic acid components of Jacobaea cannabifolia (Less.) on inflammation caused by PM2.5.

PM2.5 is an important environmental problem threatening human health at present, which poses serious harm to human body after inhalation. J. cannabifolia is a traditional Chinese medicine which exhibits anti-inflammatory effect. This study aimed to investigate the inhibitory effect of main phenolic acid components of J. cannabifolia on inflammation caused by PM2.5. Effect of PM2.5 on cell activity and apoptosis were determined by MTT, flow cytometry and calcein AM/PI staining. PHBA, PHPAA, and mixture of PHBA and PHPAA of different concentrations were given to RAW264.7 cells pretreated with PM2.5. The effect of drugs on cellular inflammatory factors was detected by ELISA. The expressions of TLRs related signal pathway at protein and gene levels were detected by western blot and qRT-PCR. The results showed that PM2.5 had no effect on cell activity and apoptosis within the determined concentration range. PHBA and PHPAA could markly inhibit the level of IL-1ß, IL-6, and TNF-α in RAW264.7 cells. Furthermore, the expressions of TLR2, TLR4, MyD88, IRAK1, TRAF6, TAK1, IKKß, and NF-κB induced by PM2.5 were markedly inhibited by PHBA and PHPAA at protein and gene levels. This study demonstrated that PHBA and PHPAA could attenuated inflammation caused by PM2.5 through suppressing TLRs related signal pathway.

Caveolin-1 contributes to realgar nanoparticle therapy in human chronic myelogenous leukemia K562 cells.

Chronic myelogenous leukemia (CML) is characterized by the t(9;22) (q34;q11)-associated Bcr-Abl fusion gene, which is an essential element of clinical diagnosis. As a traditional Chinese medicine, realgar has been widely used for the treatment of various diseases for >1,500 years. Inspired by nano-drug, realgar nanoparticles (NPs) have been prepared with an average particle size of <100 nm in a previous work. Compared with coarse realgar, the realgar NPs have higher bioavailability. As a principal constituent protein of caveolae, caveolin-1 (Cav-1) participates in regulating various cellular physiological and pathological processes including tumorigenesis and tumor development. In previous studies, it was found that realgar NPs can inhibit several types of tumor cell proliferation. However, the therapeutic effect of realgar NPs on CML has not been fully elucidated. In the present paper, it was demonstrated that realgar NPs can inhibit the proliferation of K562 cells and degrade Bcr-Abl fusion protein effectively. Both apoptosis and autophagy were activated in a dose-dependent manner in realgar NPs treated cells, and the induction of autophagy was associated with class I phosphoinositide 3-kinase/protein kinase B/mammalian target of rapamycin pathway. Morphological analysis indicated that realgar NPs induced differentiation effectively in CML cells. Furthermore, it was identified that Cav-1 might play a crucial role in realgar NP therapy. In order to study the effects of Cav-1 on K562 cells during realgar NP treatment, a Cav-1 overexpression cell model was established by using transient transfection. The results indicated that Cav-1 overexpression inhibited K562 cell proliferation, promoted endogenic autophagy, and increased the sensitivity of K562 cells to realgar NPs. Therefore, the results demonstrated that realgar NPs degraded Bcr-Abl oncoprotein, while the underlying mechanism might be related to apoptosis and autophagy, and Cav-1 might be considered as a potential target for clinical comprehensive therapy of CML.