壳寡糖对奶牛外周血单个核细胞抗氧化功能的促进作用

Promoting Effect of Chitooligosaccharides on Antioxidant Function of Peripheral Blood Mononuclear Cells of Dairy Cows

本试验以奶牛外周血单个核细胞(PBMC)为模型,旨在研究不同剂量壳寡糖(COS)对细胞活力、抗氧化功能和炎症因子含量的影响。采用单因素随机试验设计,将体外分离得到的PBMC随机分为5组,每组6个重复,对照组(CON组)细胞培养液中不添加COS;COS40、COS80、COS160和COS320组在细胞培养液中分别添加40、80、160和320μg/mL的COS,在培养箱中培养48 h。结果表明:体外添加COS对PBMC抗氧化功能的促进效果呈剂量效应,可增强抗氧化酶如硫氧还蛋白还原酶(TrxR)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)的活性,其中以160μg/mL的COS具有较好的效果。COS可抑制炎症因子白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)的产生,降低诱导型一氧化氮合酶(iNOS)的活性与一氧化氮(NO)的含量,并呈现剂量依赖性,以160μg/mL COS具有较强的抑制作用,而320μg/mL COS的抑制作用减弱。体外添加COS抑制了核因子-κB(NF-κB)p 50和NF-κB p 65的基因表达,并且呈现剂量依赖性。综上可知,COS可通过抑制NF-κB信号通路活性降低iNOS与炎症因子的基因表达以及NO的生成,进而提高PBMC的抗氧化功能。

In this experiment,dairy cows’peripheral blood mononuclear cells(PBMC)were used as a model to study the effects of different doses of chitooligosaccharides(COS)on cell viability,antioxidant function and inflammatory factor contents.The test adopted a single-factor randomized test design,and the isolated PBMC in vitro were randomly divided into 5 groups with 6 replicates per group,one was the control group(CON group),and COS was not added to the cell culture medium;the remaining 4 groups were COS40,COS80,COS160 and COS320 groups,in which,the 40,80,160 and 320μg/mL COS were added to the cell culture medium,respectively.The PBMC were incubated for 48 h in incubator.The results showed that the in vitro addition of COS had a dose-responsive effect on the antioxidant function of PBMC,which enhanced the activities of antioxidant enzymes thioredoxin reductase(TrxR),catalase(CAT)and superoxide dismutase(SOD).Among them,COS with 160μg/mL had a better effect.COS inhibited the production of inflammatory factors interleukin-1β(IL-1β),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α),and reduced the inducible nitric oxide synthase(iNOS)activity and nitric oxide(NO)content in a dose-dependent model.It had a good reduction effect in the COS dose of 160μg/mL,and the inhibitory effect of 320μg/mL COS was weakened.The in vitro addition of COS inhibited the gene expression of nuclear factor-κB(NF-κB)p 50 and NF-κB p 65,and in a dose-dependent model.It is suggested that COS may reduce the gene expression of iNOS and inflammatory factors and the production of NO by inhibiting the activity of NF-κB signaling pathway,and then promotes the antioxidant function of PBMC.