A robust in vitro screening assay to identify NF-kappaB inhibitors for inflammatory muscle diseases.
A robust in vitro screening assay to identify NF-kappaB inhibitors for inflammatory muscle diseases.
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DOI：
10.1016/j.intimp.2009.07.001
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发表时间：
2009-09
影响因子：
5.6
通讯作者：
Nagaraju K
中科院分区：
文献类型：
--
作者：
Baudy AR;Saxena N;Gordish H;Hoffman EP;Nagaraju K
具体的治疗方法是不适用于炎症性肌肉疾病。我们和其他人已经表明，在这些条件下，促炎性NF-κB通路高度活化。由于NF-κB是一个重要的治疗靶点，因此我们决定利用体外筛选试验来鉴定在稳定表达NF-κB荧光素酶报告基因的C2 C12肌系中阻断TNF-α诱导的NF-κB活化的潜在抑制剂。我们已经在未分化的成肌细胞和分化的肌管中测试了多种潜在的抗炎剂，并发现每种细胞类型的抑制强度不同。有趣的是，我们发现一些已知抑制免疫细胞中NF-κB的药物在肌肉细胞中无效。采用MTT细胞活力测定法评价药物毒性，并通过成肌细胞中NF-κB核转位的免疫染色验证荧光素酶测定法的有效性。总之，我们已经确定了最佳的检测条件，用于检测潜在的有价值的NF-κB抑制剂的第一次在肌肉细胞系，可能有显着的治疗潜力的炎症性肌肉疾病。
Specific therapies are not available for inflammatory muscle diseases. We and others have shown that the pro-inflammatory NF-κB pathway is highly activated in these conditions. Since NF-κB is an important therapeutic target, we decided to utilize an in vitro screening assay to identify potential inhibitors that block TNF-α induced NF-κB activation in a C2C12 muscle line that stably expresses an NF-κB luciferase reporter gene. We have tested multiple potential anti-inflammatory agents in undifferentiated myoblasts as well as differentiated myotubes and found different strengths of inhibitions in each cell type. Interestingly, we found that some drugs that are known to inhibit NF-κB in immune cells were not effective in muscle cells. Drug toxicity was assessed for using an MTT cell viability assay, and the validity of the luciferase assay was verified by immunostaining for NF-κB nuclear translocation in myoblasts. In conclusion, we have determined the optimal assay conditions for detecting potentially valuable NF-κB inhibitors for the first time in a muscle cell line that may have significant therapeutic potential for inflammatory muscle diseases.
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影响因子：
2.7
作者：
Waelchli, R;Bollbuck, B;Schlapbach, A
通讯作者：
Schlapbach, A
DOI：
10.1046/j.1365-201x.2001.00823.x
发表时间：
2001-03-01
期刊：
ACTA PHYSIOLOGICA SCANDINAVICA
影响因子：
--
作者：
Nagaraju, K
通讯作者：
Nagaraju, K
影响因子：
2.9
作者：
SARABIA, V;RAMLAL, T;KLIP, A
通讯作者：
KLIP, A
影响因子：
9.9
作者：
Chen, YW;Nagaraju, K;Hoffman, EP
通讯作者：
Hoffman, EP
影响因子：
56.9
作者：
AUPHAN, N;DIDONATO, JA;KARIN, M
通讯作者：
KARIN, M