Integrating 16S sequencing and metabolomics study on anti-rheumatic mechanisms against collagen-induced arthritis of Wantong Jingu Tablet / 中国天然药物

Wantong Jingu Tablet (WJT), a mixture of traditional Chinese medicine, was reported to relieve the symptoms of rheumatoid arthritis (RA), but its pharmacological mechanism was not completely understood. The aim of this study was to investigate the therapeutic mechanisms of WJT for RA in vivo. The effects of WJT on joint pathology, as well as the levels of Bax, Bcl-2, caspase-3, cleaved-caspase-3, ERK1/2, pERK1/2, TNF-α, IL-1β, and IL-6 were measured using collagen-induced arthritis (CIA) rats. The intestinal flora composition and the metabolites alteration were analyzed by 16S rDNA sequencing and metabolomics method, respectively. We found that WJT ameliorated the severity of the CIA rats which might be mediated by inducing apoptosis, inactivating the MEK/ERK signals and reducing the production of pro-inflammatory cytokines. WJT, in part, relieved the gut microbiota dysbiosis, especially bacterial phylum Bacteroidetes, Tenericutes and Deferribacteres, as well as bacterial genus Vibrio, Macrococcus and Vagococcus. 3'-N-debenzoyl-2'-deoxytaxol, tubulysin B, and magnoline were significantly associated with the specific genera. We identified serotonin, glutathione disulfide, N-acetylneuraminic acid, naphthalene and thromboxane B2 as targeted molecules via metabolomics. Our findings contributed to the understanding of RA pathogenesis, and WJT played essential roles in gut microbiota health and metabolite modulation in the CIA rats.

Sorafenib and octreotide combination therapy can inhibit proliferation of and induce apoptosis in human hepatoma cells / 中华肝脏病杂志

To investigate the effects of sorafenib and octreotide combination treatment on cellular proliferation and explore the underlying molecular mechanisms by using an in vitro cell culture system with the human hepatoma cell line, HepG2. HepG2 cells were treated with different concentrations of sorafenib and octreotide alone or in combination. Untreated HepG2 cells were used as controls. Treatment-induced cytotoxicity was determined with the cell counting kit-8 by Sigma-Aldrich, and rate of apoptosis was detected by flow cytometry. Fluorescent microscopy was used to observe rates of cell growth under the various treatments. Treatment-induced changes in protein expressions were detected by enzyme-linked immunosorbent assay (for vascular endothelial growth factor (VEGF)) and Western blotting (for the Mcl-1 apoptosis mediator and the ERK1/2 and PERK1/2 kinases). Sorafenib and octreotide, used alone or in combination, inhibited proliferation and induced apoptosis in HepG2 cells. Combination treatment was more effective than either mono-treatment (F = 200.398, P less than 0.05). Fluorescent microscopy showed that combination treatment stimulated phosphatidylserine, the marker of early apoptosis, better than either mono-treatment. VEGF expression in cultures exposed to combination treatment was also significantly lower than in mono-treatment or untreated control cultures (F = 1019.725, P less than 0.05). Western blotting showed that octreotide mono-treatment had no effect on Mcl-1 expression (vs. control group; P more than 0.05) and that combination treatment significantly lowered Mcl-1 expression (vs. mono-treatment and control groups; P less than 0.05). None of the treatments affected ERK1/2 expression (all, P more than 0.05), while all treatments significantly lowered PERK1/2 expression (vs. control group; F = 2.401, P less than 0.05) and the combination treatment lowered PERK1/2 significantly more than either mono-treatment (P less than 0.05). Sorafenib and octreotide can inhibit proliferation and induce apoptosis in the human hepatoma cell line, HepG2. Combination treatment is significantly more efficacious (P less than 0.05) and produced synergistic effects. The mechanism underlying this phenomenon may depend on synergistic inhibition of VEGF, the anti-apoptotic protein Mcl-1, and the proliferation-inducing PERK1/2.

The study of correlation between anti-cyclic citrnllinated peptide antibody and rheumatoid arthritis / 中华风湿病学杂志

Objective To explore the correlation between anti-cyclic citrullinated peptide(A-CCP) antibody and tumor necrosis factor(TNF)-?, rheumatoid factor(RF), ESR, PLT count and clinical features in patients with rheumatoid arthritis(RA), and the outcome of unclassified arthritis(arthralgia)patients after six months follow up. The value of A-CCP antibdy in the diagnosis of early RA and its pathogenetic roles is in- vestigated. Methods A-CCP antibody and TNF-?were detected by ELISA and the RF was tested by the rate scatting immunity method in 91 RA patients, 46 unclassified arthritis(arthralgia)patients and 45 other rheumatic diseases patients. Results A-CCP antibody levels in serum correlated significantly with TNF-?levels, PLT count and the degree of joint swelling in RA and unclassified arthritis(arthralgia)patients(r= 0.854, P=0.O00; r=0.882, P=0.000; r=0.318, P=0.002; r=0.486, P=0.001; r=0.291, P=0.005; r=0.731, P= 0.000 respectively). A-CCP antibody levels in serum was weakly negatively correlated with the gripping power in RA patients(r=0.228, P=0.030). And it was weakly correlated with ESR in unclassified arthritis(arthrai- gia)patients(r=0.365, P=0.013). Compared with other rheumatic diseases patients, A-CCP antibody levlels in serum increased significantly in RA and unclassified arthritis(arthralgia)patients(P=0.000). Compared with normal controls, it increased in other rheumatic diseases patients(P=0.011). Twenty-four patients had positive A-CCP antibody in 46 unclassified arthritis(arthralgia)patients. Thirty-two out of 46 unclassified arthritis(arthralgia)patients were early RA after 6 monthes follow up. 95.8%(23/24)unclassified arthritis (arthralgia)patients with positive A-CCP antibody were early RA. Conclusion A-CCP antibody reflects disease activity in certain extent. It's benefit to the diagnosis of early RA. High A-CCPantibody levels com- bined with high levels of TNF-?, ESR, PLT count and joint swelling can help the diagnosis of early RA.