The Effect Of SelW On Apoptosis In Liver Cells Of Chicken

Selenoprotein (SelW), one of selenprotein families, can express in many tissue especially liver.SelW was served as an antioxidant in chicken at present; however, it is unclear how SelW play arole to protect liver of chicken against oxidation stress. Our research was researched in two respects.In vivo which chicken were fed Se-deficient granulated diet, containing0.032mg/kg Se for15,25,35,45,55days. The liver tissues were examined for inflammation factors (cox-2, PTGEs, TNF-α,iNOS, NF-кB), cytokine (IL-1, IL-4, IL-6, IL-8, IL-10, IL-17, IFN-γ) and apoptosis (P53,caspase-3, Bax, Bak-1, Bcl-2) mRNA and protein by real-time quantitative reverse transcriptionPCR (qPCR) and western blot. In vitro overexpression and interfering of SelW liver cells wereconstructed, then we monitored the extent of the H2O2-induced necrosis and detected the abundanceof inflammation, cytokines and apoptosis mRNA and protein by qPCR and western blot. Theresults were displayed as follows:1The liver cells of SelW overexpression and knocked down were successfully constructed.The pcDNA3.1/GFP was transfected in liver cells, and the ratio of translation reagent andpcDNA3.1/GFP was8:2(μL:μg), the efficiency of translation was the highest, and higher2000-foldthan control group. Moreover, a high transfection was observed in primary cultured chicken liver24h after the Stealth RNA (S-RNA) transfection, and the optimal condition for transfection withS-RNA are:1.2μL S-RNA along with1μL LipofectamineTM RNAiMAX for six-well plate. AndS-RNA sequence has a highest gene knockdown for SelW mRNA in chicken, and the expression ofSelW was lower50%than control group.2In the chicken of Se-deficiency, the expression of P53, caspase-3, Bax and Bak-1mRNAwere higher than control group, and the Bcl-2was lower. In the overexpression SelW group, theexpression of P53, caspase-3,Bax and Bak-1mRNA were higher than control group, and theexpression of Bcl-2mRNA was higher and then lower with increasing of time. When theexpression of SelW was higher, the expression of P53、caspase-3、Bax、Bak-1mRNA were lowerthan control group, however, the expression of Bcl-2was higher, we found the death rate was lowerin overexpression SelW than in knockdown. It declared that the SelW can develop anti-apoptosisand against apoptosis to protect liver from oxidation stress.3In the chicken of Se-deficiency, the expression of cox-2、PTGEs、TNF-α、iNOS and NF-кBmRNA were higher than control group. In the overexpression SelW group, the expression of cox-2、PTGEs、TNF-α、iNOS and NF-кB mRNA were higher than control group. When the expression of SelW was higher, the expression of cox-2, PTGEs, TNF-α, iNOS and NF-кB mRNA were lowerthan control group. It declared that the SelW can develop anti-inflammations and againstinflammations to protect liver.4In the chicken of Se-deficiency, the expression of When the expression of IL-1、IL-6、IL-8and IL-17mRNA were higher than control group, and IL-4, IL-10and IFN-γ were lower. In theoverexpression of SelW group, the expression of IL-1、IL-6、IL-8and IL-17mRNA were higherthan control group, and the expression of IL-4and IL-10mRNA were higher and then lower withincreasing of time, the expression of IFN-γ was lower. When the SelW was knocked down, theexpression of IL-1、IL-6、IL-8and IL-17mRNA were lower than control group, and the expressionof IFN-γ was higher. However, the expression of Bcl-2was higher; we found the death rate waslower in overexpression SelW than in knockdown. It declared that the SelW can modulate thecytokines expression to develop immunity of chicken.