Daphnetin combined with 5-aza-Dc and a Bcl-2-miRNA interference plasmid induces apoptosis in collagen-induced arthritic rat synovial cells
详细信息 查看官网全文
- 英文论文题名：Daphnetin combined with 5-aza-Dc and a Bcl-2-miRNA interference plasmid induces apoptosis in collagen-induced arthritic rat synovial cells
- 论文作者：Li yanyan ; Zeng xiaoping ; Chen xiaoying ; Zheng mao ; Kuang nanzhen ; Fu yingyuan
- 英文论文作者：Li yanyan ; Zeng xiaoping ; Chen xiaoying ; Zheng mao ; Kuang nanzhen ; Fu yingyuan ; Nanchang University
- 年：2016
- 作者机构：Nanchang University;
- 英文论文关键词：Daphnetin ; Acute toxicity test ; Ames test ; Micronucleus assay ; Acute skin allergy test ; Local mucosa
- 会议召开时间：2016-11-04
- 会议录名称：第十一届全国免疫学学术大会摘要汇编
- 英文会议录名称：Abstracts of 2016 CSI Congress on Immunology
- 语种：英文
- 分类号：R593.22
- 学会代码：IGMD
- 会议名称：第十一届全国免疫学学术大会
- 会议地点：中国安徽合肥
- 主办单位：中国免疫学会
- 学会名称：中国免疫学会
- 页数：1
- 文件大小：973k
- 原文格式：D
- 会议级别：全国
摘要
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 cytokines.Objective: 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 CIA-FLSs.Results: 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 apoptosis.Conclusion: 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.
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 cytokines.Objective: 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 CIA-FLSs.Results: 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 apoptosis.Conclusion: 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.
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 cytokines.Objective: 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 CIA-FLSs.Results: 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 apoptosis.Conclusion: 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.
引文