Effect of 4-HDHA 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 performed lipidomics sequencing of TNF-a treated cell line mimicking the colitis in vivo.Surprisingly, we found that 4-HDHA,an autoxidation product of docosahexaenoic acid (DHA),had anti-apoptosis effect on colon epithelial cell line.To evaluate the underlying mechanism of the specific lipid perioxidin's anti-apoptosis effect, we performed the experiment using high-throughput sequencing and get data to explain the reason why 4-HDHA could decrease TNF-induced cell apoptosis. Overall design: To investigate the reason why 4-HDHA could rescue TNF-a induced cell death, we use human recombinate TNF-a to induce HT-29 cell apoptosis.At the same time, we treat the TNF-a induced cell line with 4-HDHA (the concentration is 600nm)for 72h.To exclude the effect of 4-HDHA on normal steady state cell, we treat the normal cell line with 4-HDHA(the same concentration as TNF-a treated group)Then we collect cells to extract RNA and perform sequencing for next step.We perform gene expression profiling analysis using data obtained from RNA-seq of 2 control groups and 2 treated groups:control,control+4-HDHA,TNF-a,TNF-a +4-HDHA.Each group has 3 technical replicates.