Preventive Effect of Daiokanzoto (TJ-84) on 5-Fluorouracil-Induced Human Gingival Cell Death through the Inhibition of Reactive Oxygen Species Production

Abstract

Daiokanzoto (TJ-84) is a traditional Japanese herbal medicine (Kampo formulation). While many Kampo formulations have been reported to regulate inflammation and immune responses in oral mucosa, there is no evidence to show that TJ-84 has beneficial effects on oral mucositis, a disease resulting from increased cell death induced by chemotherapeutic agents such as 5-fluorouracil (5-FU). In order to develop effective new therapeutic strategies for treating oral mucositis, we investigated (i) the mechanisms by which 5-FU induces the death of human gingival cells and (ii) the effects of TJ-84 on biological events induced by 5-FU. 5-FU-induced lactate dehydrogenase (LDH) release and pore formation in gingival cells (Sa3 cell line) resulted in cell death. Incubating the cells with 5-FU increased the expression of nucleotide-binding domain and leucine-rich repeat containing PYD-3 (NLRP3) and caspase-1. The cleavage of caspase-1 was observed in 5-FU-treated cells, which was followed by an increased secretion of interleukin (IL)-1β. The inhibition of the NLRP3 pathway slightly decreased the effects of 5-FU on cell viability and LDH release, suggesting that NLRP3 may be in part involved in 5-FU-induced cell death. TJ-84 decreased 5-FU-induced LDH release and cell death and also significantly inhibited the depolarization of mitochondria and the up-regulation of 5-FU-induced reactive oxygen species (ROS) and nitric oxide (NO) production. The transcriptional factor, nuclear factor-κB (NF-κB) was not involved in the 5-FU-induced cell death in Sa3 cells. In conclusion, we provide evidence suggesting that the increase of ROS production in mitochondria, rather than NLRP3 activation, was considered to be associated with the cell death induced by 5-FU. The results also suggested that TJ-84 may attenuate 5-FU-induced cell death through the inhibition of mitochondrial ROS production.

Impact of the use of Kampo medicine in patients with esophageal cancer during chemotherapy:a clinical trial for oral hygiene and oral condition

“…In a previous in vitro study, we showed that Daiokanzoto attenuates 5 -FU-induced cell death through the inhibition of mitochondrial ROS production (12). However, in this study we could not show any preventive effect of Daiokanzoto on the onset of oral mucositis.…”

“…Hangeshashinto was effective for the treatment of oral ulcer (4) and it is covered by the Japanese insurance. In addition, it demonstrated in our previous study that Daiokanzoto possessed the preventive effect on 5 -FU induced human gingival cell death (12).…”

The Daiokanzoto (TJ-84) Kampo Formulation Reduces Virulence Factor Gene Expression in Porphyromonas gingivalis and Possesses Anti-Inflammatory and Anti-Protease Activities

“…Yoshida et al [ 37 ] recently showed that Daiokanzoto can attenuate 5-fluorouracil-induced cell death by inhibiting the production of mitochondrial reactive oxygen species. This led to the suggestion that Daiokanzoto could be used to treat oral mucositis in patients receiving multicycle chemotherapy.…”

Synergism of co-delivered nanosized antioxidants displayed enhanced anticancer efficacy in human colon cancer cell lines

“…After treatment, cells were incubated with 5 μM JC-1 (ENZO life sciences) stain for 30 min at 37 °C, washed and images were visualized by fluorescence microscope [59] (Nikon Eclipse 80i equipped with Nikon DS-Ri1 12.7 megapixel camera, Japan). Fluorescent intensity per cell in four image frames for each group was quantified using NIH ImageJ analysis software (USA) by following reports [60] , [61] , [62] , [63] . Mitochondrial depolarization in each group was quantified by calculating the ratio of red to green fluorescent intensity and was normalized to the control (red-to-green fluorescent ratio of control considered as 1) [62] , [63] , [64] .…”