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An understanding gene-gene interaction helps users to design the next experiments efficiently and (if applicable) to make a better decision of drugs application based on the different biological conditions of the patients. This study aimed to identify changes in the hidden relationships between pro- and anti-inflammatory cytokine genes in the bovine oviduct epithelial cells (BOECs) under various experimental conditions using a multilayer response surface method. It was noted that under physiological conditions (BOECs with sperm or sex hormones, such as ovarian sex steroids and LH), the mRNA expressions of  IL 1 0 ,  IL1B ,  TNFA ,  TLR4 , and  TNFA  were associated with  IL1B ,  TNFA ,  TLR4 ,  IL4 , and  IL10 , respectively. Under pathophysiological + physiological conditions (BOECs with lipopolysaccharide + hormones, alpha-1-acid glycoprotein + hormones, zearalenone + hormones, or urea + hormones), the relationship among genes was changed. For example, the expression of  IL10  and  TNFA  was associated with ( IL1B ,  TNFA , or  IL4 ) and  TLR4  expression, respectively. Furthermore, under physiological conditions, the co-expression of  IL10  +  TNFA ,  TLR4  +  IL4 ,  TNFA  +  IL4 ,  TNFA  +  IL4 , or  IL10  +  IL1B  and under pathophysiological + physiological conditions, the co-expression of  IL10  +  IL4 ,  IL4  +  IL10 ,  TNFA  +  IL10 ,  TNFA  +  TLR4 , or  IL10  +  IL1B  were associated with  IL1B ,  TNFA ,  TLR4 ,  IL10 , or  IL4  expression, respectively. Collectively, the relationships between pro- and anti-inflammatory cytokine genes can be changed with respect to the presence/absence of toxins, sex hormones, sperm, and co-expression of other gene pairs in BOECs, suggesting that considerable cautions are needed in interpreting the results obtained from such narrowly focused  in vitro  studies.

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Understanding the hidden relations between pro- and anti-inflammatory cytokine genes in bovine oviduct epithelium using a multilayer response surface method