Effect of ferrostatin-1 and liproxstatin-1 during TNF-induced HT-29 cells apoptosis

Ulcerative colitis (UC) is an inflammatory disease characterized by an uncontrolled inflammatory response. Previous study showed that the immunological impairment elicted the alteration of inflammatory mediators, and ferroptosis was implicated with the lethal accumulation of reactive oxygen species (ROS).We have already found that lipid ROS level was elevated in both murine DSS colitis model and TNF-a-induced cell apoptosis model in vitro.Then we here use two ferroptosis inhibitors ferrostatin-1 and liproxstatin-1 trying to eliminate lipid ROS in order to relive the symptoms of colitis.But we strikingly notice that liproxstatin-1 could increase TNF-induced cell apoptosis instead of protecting cells from death as we expected before while the other inhibitor ferrostatin-1 could protect cells from death indeed.To evaluate the underlying mechanism, we performed the experiment using high-throughput sequencing and get data to explain the reason why liproxstatin-1 could increase TNF-induced cell apoptosis . To investigate the reason for different effects of the two ferroptosis inhibitors ferrostatin-1 and liproxstatin-1 during TNF-induced HT-29 cells apoptosis，we firstly use human Recombinate TNF-a to induce HT-29 cell apoptosis.At the same time, we treat the TNF-a induced cell line using 20um ferrostatin-1 or 20um liproxstatin-1 for 72h.Then we collect cells to extract RNA and perform sequencing for next step.Notably,we also add 20um ferrostatin-1 or 20um liproxstatin-1 to the normal cell line which is not treated by TNF-a for 72h to exclude the effects of ferrostatin-1 and liproxstatin-1 on steady state cell.We then perform gene expression profiling analysis using data obtained from RNA-seq of 3 control groups and 3 treated groups:control,control+ferrostatin-1,control+liproxstatin-1,TNF-a，TNF-a+ferrostatin-1，TNF-a+liproxstatin-1.Each group has 3 technical replicates.