Shikonin (Anchusa acid) is a natural product, a TMEM16A chloride channel inhibitor (IC50=6.5 μM) and selective PKM2 inhibitor. Shikonin exhibits antitumor, anti-inflammatory and wound healing activities.

PKM2 (FBP absence):0.3 μM, PKM2 (FBP presence):0.8 μM, TMEM16A (Cl- channel):6.5 μM

方法：人胶质瘤细胞 U87 和 U251 用 Shikonin (2.5-7.5 μmol/L) 处理 12-72 h，使用 CCK-8 方法检测细胞活力。
结果：Shikonin 以时间和剂量依赖的方式抑制 U87 和 U251 细胞的增殖。[1]
方法：巨噬细胞 RAW264.7 用 Shikonin (0.5-2 μM) 预处理 1 h，然后用 LPS (250 ng/mL) 和 IFN-γ (100 ng/mL) 处理 24 h，使用 RT-qPCR 和 ELISA 检测炎症因子水平。
结果：在 LPS+IFN-γ 模拟的 RAW264.7 细胞中，IL-1β、IL-6 和 TNF-α 的 mRNA 和蛋白表达均降低。[2]

方法：为研究对小鼠结肠炎的疗效，将 Shikonin (6.125-25 mg/kg) 灌胃给药给 DSS 诱导的 Balb/c 小鼠结肠炎模型，每天一次，持续七天。
结果：Shikonin 减轻了 DSS 诱导的小鼠结肠炎的总体症状，并减轻了结肠损伤。[2]
方法：为检测体内抗肿瘤活性，将 Shikonin (0.1-10 mg/kg) 腹腔注射给携带人黑色素瘤 B16 的 SCID 小鼠，每天一次，持续九天。
结果：Shikonin 处理以剂量依赖的方式抑制 SCID 小鼠中的 B16 细胞生长。[3]

U87 and U251 cells are seeded into 96-well plates at a density of 1×104 cells per well in standard DMEM and incubated for 24 h under standard conditions (37°C and 5% CO2). Then the medium is replaced with either blank, serum-free DMEM or DMEM containing Shikonin at concentrations of 2.5, 5, and 7.5 μM. The total volume in each well is 200 μL. Finally, the plates are shaken softly and the optical density is recorded at 570 nm (OD570) using a plate reader. At least three independent experiments are performed[4].

Healthy male Sprague-Dawley rats (n=30; 8 to 10-weeks old, 250 to 300 g) are used in this study. Rats were randomly assigned to three groups: Sham-operated group (n=10), osteoarthritis model group (n=10) and Shikonin-treated group (n=10). In the sham-operated group, the right knee joint of the anesthetized rat is only exposed under sterile conditions, and the rats are treated with 0.1 ml/100 g physiological saline (i.p.). In the osteoarthritis model group, osteoarthritis model rats were treated with 0.1 ml/100 g physiological saline (i.p.). In the Shikonin-treated group, osteoarthritis model rats are treated with 10 mg/kg Shikonin (i.p.) once daily for 4 days after osteoarthritis modeling[5].