JSH-23 is an NF-κB inhibitor that inhibits NF-κB transcriptional activity (IC50=7.1 μM) but does not affect IκBα degradation. JSH-23 is an antioxidant with anti-inflammatory activity.

NF-κB:7.1 μM

方法: 骨髓巨噬细胞 BMMs 用 JSH-23 (0.78-200 µM) 处理 48-96 h，通过 CCK-8 assay 检测细胞活力。
结果: JSH-23 在 50 µM 以下的浓度下没有可检测到的毒性作用。[1]
方法: 携带报告基因 pNF-κB-SEAP-NPT 的巨噬细胞 RAW 264.7 用 LPS (1 µg/mL) 和 JSH-23 (1-30 µM) 处理 16 天，检测 SEAP 表达以反映 NF-κB 转录活性。
结果: JSH-23 以剂量依赖的方式抑制 LPS 诱导的 SEAP 表达，3 µM 时抑制 23±3%，10 µM 时 68±3%，30 µM 时 103±4%。JSH-23 抑制 NF-κB 转录活性。[2]

方法: 为研究在糖尿病神经病变中的作用，将 JSH-23 (1-3 mg/kg) 口服给药给 streptozotocin 诱导的糖尿病大鼠，每天一次，持续两周。
结果: JSH-23 治疗显著逆转了糖尿病动物的神经传导和神经血流缺陷。治疗也部分纠正了机械痛阈的降低。JSH-23 处理可抑制坐骨神经中 p65/p50 亚基的核转位。[3]

Measurement of NF-κB transcriptional activity: Macrophages RAW 264.7 transfected stably with reporter plasmid of pNF-κB-SEAP-NPT are treated with 1 μg/ml LPS and/or sample for 16 hours. As the reporter, SEAP activity in the cell-free culture media is measured as followed. Single cell-derived stable transfectants are plated in 5 ml of T-25 flask, and the media is decanted 24 h later. At this time, cells are washed twice with phosphate-buffered saline, and incubations are initiated by addition of new media. Chemicals are added to the culture medium after 24 h of incubations. Aliquots (25 ml) of medium from a control or chemical-treated cultures are taken at 0, 3, 20, 24, 48, and 72 h, heated at 65°C for 5 min to eliminate the alkaline phosphatase activity, and used immediately or stored at -20°C. Mixtures consisting of dilution buffer (25 ml), assay buffer (97 ml), culture media (25 ml), and 4-methylumbelliferyl phosphate (MUP, 1 mM, 3 ml) in each well of the 96-well plates are incubated for 60 min in the dark at room temperature. Fluorescence emits the product of the SEAP/MUP is measured at 449 nm using a 96-well plate fluorometer after excitation at 360 nm.

Macrophages RAW 264.7 are incubated with various concentrations of JSH-23 compound for 24 h. The cells are treated with WST-1 solution and absorbance is measured at 450 nm.(Only for Reference)