Cell experiment:

The assay is performed following a standard published procedure using microtiter plate format and sulforhodamine B standing at the end point. The leukemic cells (P388) are exposed to Flaconitine (3-Acetylaconitine) for three days. Cells are grown and maintained in an incubator set at 37℃, 5% CO2 and 95% humidity. RPMI-1640 media supplemented with 10% fetal calf serum (FCS) and 0.5% trypsin are used to culture P388 cells[2].

Flaconitine is isolated from the ammonium hydroxide wetted root of A.szechenyianum Gay. Flaconitine is considered to be a NF-κB inhibitor.

Flaconitine has been demonstrated to suppress TNF-α induced NF-κB activation and is considered as a diester-diterpenoid-type NF-κB inhibitor[1] Flaconitine (3-Acetylaconitine) is evaluated for its cytotoxic activity against human leukemic P388 cells using the SRB (Sulforhodamine B) method, with Etoposide as the positive control. The cell growth inhibitory potency of Flaconitine is expressed as IC50 (μM) Value. Flaconitine exhibits moderately cytotoxic activity with an IC50 of 18.6 μM[2].

[1]. Dong L, et al. Identification of nuclear factor-κB inhibitors and β2 adrenergic receptor agonists in Chinese medicinal preparation Fuzilizhong pills using UPLC with quadrupole time-of-flight MS. Phytochem Anal. 2014 Mar-Apr;25(2):113-21. [2]. He YQ, et al. Diterpenoid alkaloids from a Tibetan medicinal plant Aconitum richardsonianum var. pseudosessiliflorum and their cytotoxic activity. J Pharm Anal. 2011 Feb;1(1):57-59.