Effects Of IL-6 And TNF-α On The Proliferation Of Skin Fibroblasts In Liaoning Cashmere Goats

Liaoning cashmere goat is a unique breed with extremely high cashmere production in China,and it is famously known as the "soft gold".The growth of Liaoning cashmere goats is affected by diverse factors,such as environment,metabolism,and genes.In recent years,more scholars have begun to study how to improve the yield and the quality of the Liaoning cashmere goats;however,currently,there are few studies on the mechanism of various factors regulating the growth of the Liaoning cashmere goat hair.Previous laboratory research indicated that After the treatment of Liaoning cashmere goat skin fibroblast 72 H with 0.2g/L melatonin,the most differentially expressed genes related to villi growth in the cells included the expression levels of IL-6 and TNF-α,which was a significant increase.IL-6 and TNF-α genes can participate in wound healing and villi growth.In addition,they can also participate in the regulation of NF-κB and TNF signaling pathways related to villi growth.Therefore,the purpose of this experiment is to associate the two genes,IL-6 and TNF-α,with the Liaoning cashmere goat skin fibroblasts,and explore the proliferation of the Liaoning cashmere goat skin fibroblasts using IL-6 and TNF-α genes.To influence the situation,we explored the mechanism of melatonin’s regulation of villi growth at the molecular and cellular levels,and provided data support for further improving the production and quality of cashmere.In the experimental design process,first of all,bioinformatics analysis was used to analyze the physical and chemical properties,glycosylation and phosphorylation site analysis,transmembrane domain,and signal peptide prediction of the proteins encoded by the two genes,IL-6 and TNF-α.Structural prediction,etc.thereafter explored the effect of 0.2 g/L melatonin on the proliferation of the Liaoning cashmere goat skin fibroblasts through the CCK-8 experiment.Subsequently,we used gene overexpression,CCK-8 and other experiments to explore the effects of overexpression of IL-6 and TNF-α on the proliferation of Liaoning cashmere goat skin fibroblasts.Finally,the experiments of the interfering IL-6 and TNF-α on the skin fibroblasts of the Liaoning cashmere goat were investigated through gene interference,QPCR,and CCK-8 experiments effects of proliferation.The experimental results showed that 0.2 g/L melatonin treatment of the Liaoning cashmere goat skin fibroblasts was beneficial to the proliferation of the cells within 24 to 48 hours,and the treatment time inhibited the proliferation of the cells within 48 to 72 hours.Either with overexpressing the two genes,IL-6 and TNF-α,or interfering with their expression,there was no significant effect on the proliferation of the Liaoning cashmere goatskin fibroblasts;however,after interfering with the expression of IL-6 and TNF-α genes,the cell viability in the four treatment groups plus melatonin showed an overall upward trend.Therefore,we believe that under natural circumstances,IL-6 and TNF-α genes have no effect on the proliferation of the Liaoning cashmere goat skin fibroblasts.They,however,have a protective effect under the conditions of melatonin treatment.