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Daphnetin combined with 5-aza-Dc and a Bcl-2-miRNA interference plasmid induces apoptosis in collagen-induced arthritic rat synovial cells

Daphnetin combined with 5-aza-Dc and a Bcl-2-miRNA interfere...

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第十一届全国免疫学学术大会

作者： Li yanyan Zeng xiaoping Chen xiaoying Zheng mao Kuang nanzhen Fu yingyuan Nanchang University

Background: Daphnetin is an active ingredient that is extracted from Jiangxi flowers(Marginata). We had applied it in to treat fibroblast-like synoviocytes(FLSs) from collagen-induced arthritic(CIA) rats resulted in the modulation of the expression of Th17 and related ***: To investigate whether Daphnetin combined with 5-aza-Dc and a Bcl-2-miRNAinterference plasmid(MRBcl-2) induces apoptosis in CIA-FLSs and to explore the mechanisms potentially underlying the activity of this combination as a treatment for RA. Methods: The relative mRNA expression levels were detected using RT-PCR. The methylation status was measured using methylation-specific PCR. The rate of apoptosis was detected in synovial cells using the Annexin V/PI double-labelling method. The protein expression levels were detected using cell-based ELISA. Transmission electron microscopy was used to observe changes in the ultrastructure of ***: We explored the effects of a combined approach including Daphnetin, 5-aza-Dc, and MR-Bcl-2. We show that this combination down-regulated the relative mRNA and protein expression levels of Bcl-2 and up-regulated the relative mRNA and protein expression levels of Bax, Bid, Caspase-3, Caspase-8, and Caspase-9. Daphnetin cooperated with 5-aza-Dc to demethylate the proapoptotic gene Bid, which increased in expression, to promote synovial cell ***: Daphnetin cooperated with 5-aza-Dc and MR-Bcl-2 to induce apoptosis in CIA-FLSs. This effect potentially induced apoptosis-related molecular mechanisms via the down-regulation of Bcl-2 and the up-regulation of Bax and Bid. It also activated the apoptotic channels involving Caspase-3, Caspase-8, and Caspase-9 to induce apoptosis in CIAFLSs.

关键词： Daphnetin acute toxicity test Ames test Micronucleus assay acute skin allergy test Local mucosa

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Toxicological studies of daphnetin

Toxicological studies of daphnetin

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第十一届全国免疫学学术大会

作者： Wang jieying Kuang nanzhen Zheng mao Zou wenwei Zeng xiaoping Fu yingyuan Nanchang University

Background: Numerous studies have demonstrated that daphnetin(DAP) has anti-inflammatory, anti-oxidant, anti-bacterial, anti-tumour, anti-arthritic and other extensive pharmacological effects. However, its genotoxicity has been rarely ***: The safety of DAP was evaluated by an acute toxicity test using a Salmonella typhimurium / mammalian microsomal enzyme assay(Ames test), bone marrow micronucleus test, acute skin allergy test, and local mucosal irritation ***: Disposable lavage mice received a dose of 100 mg /(kg · body weight), which is more than 175 times the body effective dose;were continuously observed for 14 d;and showed no signs of poisoning. Each dose of DAP resulted in revertant colonies that were less than two times the number of mutation colonies in the negative control group, indicating that the DAP Ames test was negative. The micronucleus rate of each DAP test dose group was not significantly different from that of the negative control group(p > 0.05) but was significantly different from the cyclophosphamide(CTX) positive control group(p skin had no stimulation reaction to DAP or 10% dimethyl sulfoxide(DMSO), including no erythema, edema, or sensitization phenomenon. Rabbits were generally in good condition after dosing the oral mucosa with DAP, and no oral mucosal erythema, erosion, ulcers or other irritation reactions were ***: Within the range of tested doses, DAP did not cause any observed toxic effects, mutagenic effects, sensitization or stimulation.

关键词： Daphnetin acute toxicity test Ames test Micronucleus assay acute skin allergy test Lo

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