苦荞球蛋白酶解液对人脐静脉内皮细胞氧化损伤的保护作用

Protective effect of enzymatic hdrolysate from Tartary buckwheat globulin against the oxidative injury of human umbilical vein endothelial cells

研究苦荞球蛋白酶解液(EHBG)对oxLDL诱导的人脐静脉内皮细胞(HUVEC)氧化损伤的保护作用。苦荞球蛋白(BG)经胃-胰蛋白酶连续酶解后制备EHBG;采用oxLDL诱导HUVEC细胞氧化损伤模型,将EHBG作用于细胞,Vc为阳性对照;MTT法检测细胞存活率,测定各组细胞培养液中乳酸脱氢酶(LDH)活性、一氧化氮(NO)含量、丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性;Westernblot检测各组细胞中凝集素样氧化低密度脂蛋白受体(LOX1)和核转录因子(NF-κB)的表达情况。结果显示,与氧化损伤模型组比较,EHBG、BG和Vc均可提高oxLDL处理后HUVEC生存率,降低细胞培养液中MDA含量、LDH活性,升高SOD活性及NO含量;下调ox-LDL诱导的内皮细胞LOX1和NF-κB的表达,且与BG相比,EHBG的效果均较显著。表明EHBG具有抗HUVEC氧化损伤的作用,其机制可能与抑制oxLDL/LOX1/NF-κB信号通路有关。

Objective:The protective effects of enzymatic hydrolysate (EHBG) against oxidative injury of human umbilical vein endothelial cells (HUVEC) were investigated.Methods:Tartary buckwheat globulin (BG) was subjected to a two-stage in vitro digestion (pepsin followed pancreatin).The groups were designed as follows:normal control group,oxLDL oxidative injury model group,enzymatic hdrolysate of BG (EHBG) group (low,medium and high concentration group),BG control group,Vc positive control group.Cell survival rate was detected by MTT assay.The lactate dehydrogenase (LDH) activity,nitric oxide (NO) content,malondialdehyde (MDA) content as well as superoxide dismutase (SOD) activity in cell culture medium of each group were measured.Western blot was used to detect the expression of lectin-like oxidized LDL receptor1 (LOX1) and nuclear factor-κB (NF-κB).Results:HUVEC oxidative damage model induced by oxLDL was successfully replicated.Compared with model group,the cell vitality (OD value) was significantly increased,the content of NO was increased,the activity of SOD was increased,the content of MDA was decreased,the activity of LDH was decreased,the protein expression of LOX1 and NF-κB was down-regulated in EHBG,BG and Vc groups,and compared with BG group, the effect of EHBG group was significant.Conclusions:EHBG had obvious protective effects against the vascular endothelial cells oxidative injury induced by oxLDL in cellular level,its mechanism may be associated with inhibition of oxLDL/LOX1/NF-κB signaling pathway.