Geniposide promotes splenic Treg differentiation to alleviate colonic inflammation and intestinal barrier injury in ulcerative colitis mice.

Geniposide has been proven to have a therapeutic effect on ulcerative colitis (UC) in animals, but its potential mechanism in UC remains to be clarified. The purpose of this study was to confirm the efficacy of geniposide in UC and to investigate the possible mechanism of geniposide in UC treatment. In vivo, geniposide relieved weight loss and reduced intestinal tissue damage in UC mice. Geniposide decreased the levels of IL-1ß and TNF-α and increased IL-10 levels in the colon and serum of UC mice. Geniposide increased FOXP3 expression in the colon and the number of CD4+ FOXP3+ cells in the spleen of UC mice. BD750 abolished the above regulatory effect of GE on UC mice. In vitro, geniposide increased the number of CD4+ FOXP3+ cells in spleen cells from normal mice, decreased the levels of IL-1ß, CCL2 and TNF-α in the supernatant of LPS-treated Caco-2 cells, and decreased the protein expression of Beclin-1 and Occludin in cacO-2 cells. Epirubicin inhibited the effect of geniposide on increasing the number of CD4+ FOXP3+ cells in spleen cells, attenuated the inhibitory effect of geniposide on proinflammatory factors and attenuated the upregulation of geniposide on tight junction proteins in LPS-treated Caco-2 cells in the coculture system. In conclusion, geniposide has an effective therapeutic effect on UC. Increasing Treg differentiation of spleen cells is the mechanism by which geniposide alleviates intestinal inflammation and barrier injury in UC.

Matrix Metalloproteinase-3 induces proteoglycan degradation in gouty arthritis model.

BACKGROUND: Gout is an inflammatory arthritis resulting from precipitation of monosodium urate (MSU) crystals in joints and surrounding tissues. However, the mechanism underlying high levels of uric acid inducing gouty arthritis has not been clarified. OBJECTIVE: The purpose was to investigate the role of Matrix Metalloproteinase-3 (MMP-3) in the development of gouty arthritis from hyperuricemia. METHOD: MSU crystal-induced gouty arthritis model and chondrocytes were used to evaluate changes of MMP-3 levels. Western blot, qPCR and ELISA were performed to detect MMP-3, Tissue Inhibitors of Metalloproteinase-1 (TIMP-1) and A Disintegrin and Metalloproteinase with Thrombospondin Motifs-4 (ADAMTS-4) expressions in rabbit chondrocytes. Expression of proteoglycan was determined through toluidine blue staining. Concentrations of glycosaminoglycan, Interleukin-6 (IL-6), Interleukin-1ß (IL-1ß) and Tumor Necrosis Factor-α (TNF-α) in chondrocytes were assessed via ELISA kits. Concentration of uric acid in supernate was tested by Automatic Analyzer. RESULTS: MMP-3 was significantly increased in rat serum, synovial fluid, cartilages and chondrocytes treated with high-level uric acid. Increased concentration of glycosaminoglycancould be observed in chondrocytes incubated with MMP-3, as well as the remarkable downregulation of proteoglycan expression. Furthermore, high-level uric acid contributed to the degradation of proteoglycan via the activation of MMP-3. IL-6, IL-1ß and TNF-α concentrations were increased significantly in 35 °C compared to 37 °C with MMP-3 and high-level uric acid. CONCLUSION: Our study showed that MMP-3 was enhanced by high levels of uric acid, which promoted proteoglycan degradation, and induced MSU crystallization in turn. A low temperature environment is an important factor in the development of gout.

Regulatory effect of melatonin on cytokine disturbances in the pristane-induced lupus mice.

Systemic lupus erythematosus (SLE) develops in relation to many environmental factors. In our opinion, it is more important to investigate the effect of melatonin on the environmental- related SLE. In the present study, 0.5 ml pristane were used to induce SLE in female BALB/c mice. Melatonin (0.01, 0.1, 1.0 mg/kg) was orally administered immediately after pristane-injection for 24 weeks. IgM anti ssDNA and histone antibodies were detected after 0, 1, 2, 4, 8 weeks pristane injection. The levels of IL-2, IL-6 and IL-13 were detected after 24 weeks. Renal lesions were also observed. The results showed that melatonin antagonized the increasing levels of IgM anti ssDNA and histone autoantibodies. Melatonin could also decrease the IL-6 and IL-13 production and increase the IL-2 production. Besides, melatonin could lessen the renal lesions caused by pristane. These results suggested that melatonin has a beneficial effect on pristane-induced lupus through regulating the cytokines disturbances.

MicroRNA­143­3p contributes to the regulation of pain responses in collagen­induced arthritis.

Patients with rheumatoid arthritis (RA) suffer from pain, which is associated with inflammation, peripheral and central pain processing, and joint structure damage. The aim of the present study was to investigate a key microRNA (miR) and its target genes that are involved in the pain responses of RA, and to clarify the mechanism of pain regulation. Collagen­induced arthritis (CIA) was induced in DBA/1 and C57BL/6 mice. The paw swelling, mechanical withdrawal threshold (MWT), thermal withdrawal latency (TWL), and expression levels of tumor necrosis factor (TNF)­α and prostaglandin (PG)E2 in the sera were investigated. Decreased MWT and TWL, and increased TNF­α and PGE2, in the CIA model group were observed in DBA/1 and C57BL/6 mice. DBA/1 mice exhibited greater hyperalgesia and higher levels of inflammatory mediators. miR­143­3p expression in the blood and the dorsal root ganglion (DRG) were detected, and low miR­143­3p expression was demonstrated in the blood and DRG tissue of CIA mice. The target genes of miR­143 were predicted and analyzed. A total of 1,305 genes were predicted and 55 pain­associated genes were obtained. Prostaglandin­endoperoxide synthase 2 (Ptgs2), MAS related GPR family member E (Mrgpre), prostaglandin D2 receptor and Tnf were selected as target genes of miR­143. DRG cells were cultured and transfected with miR­143­3p inhibitor or mimic. The expression of Mrgpre, Ptgs2 and Tnf was significantly inhibited following miR­143­3p mimic transfection, while the expression of Mrgpre, Ptgs2 and Tnf was increased following inhibitor transfection. Additionally, the expression of pain­associated genes in the DRG of mice was investigated and the expression of Ptgs2, Mrgpre and Tnf in the DRG of CIA mice was also significantly upregulated. These results revealed that CIA mice exhibited marked hyperalgesia and high levels of inflammatory pain mediators. Low expression of miR­143­3p negatively regulated the pain­associated target genes, including Mrgpre, Ptgs2 and Tnf, thereby affecting chronic inflammatory pain and neuropathic pain in RA.

The forecast of anticancer targets of cryptotanshinone based on reverse pharmacophore-based screening technology.

Anticancer targets of cryptotanshinone were evaluated and rapidly forecasted with PharmMapper, a reverse pharmacophore-based screening platform, as well as drug target databases, including PDTD, DrugBank and TTD. The pathway analyses for the collection of anticancer targets screened were carried out based on the KEGG pathway database, followed by the forecast of potential pharmacological activities and pathways of the effects of cryptotanshinone, and verification of some of the targets screened using whole cell tests. The results showed that a total of eight targets with anticancer potential were screened, including MAP2K1, RARα, RXRα, PDK1, CHK1, AR, Ang-1 R, and Kif11. These targets are mainly related to four aspects of the cancer growth: the cell cycle, angiogenesis, apoptosis, and androgen receptor. The cell tests showed that cryptotanshinone can inhibit the viability of human hepatoma cells SMMC-7721, which is related to the reduction of expression of MAP2K1 mRNA. This method provides a strong clue for the study of the anticancer effects and mechanisms of action of cryptotanshinone in the future.

Shikonin reduces endometriosis by inhibiting RANTES secretion and mononuclear macrophage chemotaxis.

Endometriosis is a common disease in females of reproductive age and has the classic characteristic of mononuclear cell infiltration into lesions. Shikonin is an anti-inflammatory phytocompound obtained from Lithospermum erythrorhizon whose potential therapeutic effects in the treatment of endometriosis remain unclear. The working hypothesis of the present study was that shikonin is capable of inhibiting the development of endometriosis by inhibiting the chemotactic effect. In a murine model of endometriosis, shikonin significantly inhibited the growth of human endometrial tissue implanted into severe combined immunodeficiency (SCID) mice (P<0.05) and no adverse effects were observed. Mouse regulated upon activation normal T-cell expressed and secreted (mRANTES) levels in the peritoneal fluid of the animal endometriosis model were higher than those in normal SCID mice (P<0.05) and decreased significantly following shikonin treatment in a dose-dependent manner (P<0.05). Peritoneal fluid from SCID mice treated with shikonin inhibited the chemotaxis of monocytes; this inhibitory effect was eradicated by mRANTES antibody. In vitro, shikonin significantly inhibited RANTES expression in U937 cells that were cultured alone or co-cultured with human mesothelial and endometrial stromal cells. In addition, shikonin inhibited the RANTES-induced chemotaxis of U937 cells (P<0.05). The results indicate that shikonin inhibits the development of endometriosis by various mechanisms, including the inhibition of RANTES expression and the reduction of mononuclear cell migration to lesions. Therefore, shikonin may be a novel, useful and safe agent for treating endometriosis.