黄连素抑制TNF-α刺激ARPE-19细胞分泌炎症因子的作用及机制研究

发布时间：2019-04-27 13:50

【摘要】：目的：黄连素（BBR）是在中国传统医药中广泛应用广谱抗菌药物。临床上主要用于治疗肠道细菌感染性疾病。它是否能影响人视网膜色素上皮细胞产生炎症因子的尚不可知。因此，研究黄连素对ARPE-19细胞产生炎症因子的影响作用及机制是本实验的目的。方法：在不同时间点，用黄连素处理与TNF-α共培养的ARPE-19细胞。酶联免疫吸附实验（ELISA）检查细胞养上清中IL-6、IL-8和MCP-1的蛋白浓度。实时定量PCR（real-time PCR）检测三种因子的mRNA水平。Western-Blot分析细胞内p38丝裂原活化蛋白激酶、胞外信号调节激酶（ERK1/2）和c-Jun N-端激酶(JNK)的磷酸化水平。使用p38、ERK1/2和JNK特异性抑制剂探讨三种因子的产生可能涉及的信号转导通路。结果：TNF-α能显著的促进ARPE-19细胞从蛋白水平以及mRNA水平产生IL-6、IL-8和MCP-1。也能提高p38、ERK1/2和JNK的磷酸化水平。而抑制剂实验表明IL-6的产生主要是通过p38路径调节，，而IL-8和MCP-1的产生与p38、ERK1/2以及JNK三条路径都有关系。黄连素则从蛋白和mRNA水平都能显著抑制由TNF-α刺激产生的IL-6、IL-8和MCP-1,并且下调p38、ERK1/2和JNK的磷酸化水平。结论：黄连素能显著抑制ARPE-19细胞表达炎症因子，并且该抑制作用是通过下调p38、ERK1/2和JNK信号通路来完成实现的。
[Abstract]:Objective: berberine (BBR) is widely used in traditional Chinese medicine. It is mainly used in the treatment of intestinal bacterial infectious diseases. It is unclear whether it can affect the production of inflammatory factors by human retinal pigment epithelial cells. Therefore, the aim of this study is to study the effect and mechanism of berberine on the production of inflammatory factors in ARPE-19 cells. Methods: ARPE-19 cells co-cultured with TNF- 伪 were treated with berberine at different time points. The protein concentrations of IL-6,IL-8 and MCP-1 in cell culture supernatant were detected by enzyme-linked immunosorbent assay (ELISA). The phosphorylation levels of p38 mitogen-activated protein kinase (p38 mitogen-activated protein kinase), extracellular signal-regulated kinase (ERK1/2) and c-Jun N-terminal kinase (JNK) were measured by real-time quantitative PCR (real-time PCR). P38, ERK 1 / 2 and JNK specific inhibitors were used to investigate the possible signal transduction pathways involved in the production of the three factors. Results: TNF- 伪 significantly promoted the production of IL-6,IL-8 and MCP-1. from protein level and mRNA level in ARPE-19 cells. The phosphorylation levels of p38, ERK 1, 2 and JNK were also increased. The inhibition experiments showed that the production of IL-6 was mainly mediated by p38 pathway, while the production of IL-8 and MCP-1 was related to p38, ERK1 / 2 and JNK pathways. Berberine could significantly inhibit IL-6,IL-8 and MCP-1, induced by TNF- 伪 and down-regulate the phosphorylation levels of p38, ERK1-2 and JNK from both protein and mRNA levels. Conclusion: berberine can significantly inhibit the expression of inflammatory factors in ARPE-19 cells, and this inhibition is achieved by down-regulating the p38, ERK-1, and JNK signaling pathways.
【学位授予单位】：重庆医科大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R285;R774

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