【作者】 黄怡； 黄琴； 李卫芬； 余东游； 周绪霞；

【Author】 HUANG Yi~(1,2),HUANG Qin~1,LI Wei-fen~(1*),YU Dong-you~1,ZHOU Xu-xia~1 (1.Institute of Feed Science,Zhejiang University,The Key Laboratory of Molecular Animal Nutrition,Ministry of Education, Hangzhou,310029,China;2.College of Animal Science and Technology,Guangxi University,Nanning 530005,China)

【机构】 浙江大学动物科学学院饲料科学研究所； 浙江大学动物分子营养学教育部重点实验室； 广西大学动物科学技术学院；

【摘要】 通过体外细胞培养的方法研究了鼠李糖乳杆菌(L.rhamnosus)对Caco-2细胞抗氧化和非特异免疫功能的影响。Caco-2细胞分别用PBS(A组)、E.coli K88(B组)和L.rhamnosus(C组)处理,以及先用L.rhamnosus预处理,再用E.coli K88处理(D组),用比浊法检测细胞培养上清的总抗氧化能力(Total antioxidative capability,T-AOC),以及细胞培养上清和细胞裂解液中的超氧化物歧化酶(Superoxide dismutase,SOD)活力;用ELISA法检测细胞培养上清的增殖诱导配体(a proliferation-inducing ligand,APRIL),IL-8和IL-10的含量。结果表明:与A组相比,B组细胞上清中T-AOC显著降低(P<0.01),SOD活力显著提高(P<0.01),但在细胞裂解液中SOD酶活力无显著变化;C组细胞上清中T-AOC显著高于A组和B组(P<0.01),但SOD则显著低于A组和B组(P<0.01),细胞裂解液中SOD活性高于A组(P<0.05)。大肠杆菌处理(处理B)能促进Caco-2细胞分泌APRIL(P<0.01),但不刺激白介素IL-8和IL-10的分泌;C组APRIL显著高于A组(P<0.01),但远低于B组(P<0.01),IL-8显著低于A和B组(P<0.01),而IL-10则显著高于A和B组(P<0.01)。与B组相比,D组APRIL显著减少(P<0.01),但IL-8和IL-10都显著增加(P<0.05和P<0.01)。以上结果说明,鼠李糖乳杆菌可提高Caco-2细胞的抗氧化功能,促进促炎细胞因子增殖诱导配体APRIL和抗炎细胞因子IL-10的分泌,抑制促炎细胞因子IL-8的产生;显著减轻肠上皮细胞对致病菌的炎症反应(APRIL显著降低),增强其免疫应答(显著提高IL-10和IL-8水平),表现出免疫保护和免疫佐剂的效果。更多还原

【Abstract】 Caco-2 cells were used to investigate the anti-oxidative and non-specific immunologic functions effects of Lactobacillus rhamnosus.Caco-2 cells were treated with PBS(group A),Escherichia coli K88(group B) and L. rhamnosus(group C),respectively,for group D,Caco-2 cells were pre-incubated with L.rhamnosus and then infected with E.coli K88.Cell culture supernatant and cell lysate were collected and analyzed.T-AOC level(in cell culture supernatant) and SOD activity(both in cell culture supernatant and cell lysate) were detected by Turbidimetry to evaluate anti-oxidative capacity of Caco-2 cells.Furthermore,the immune activity of Caco-2 cells were assessed by measuring levels of APRIL,IL-8 and IL-10 in cell culture supernatant with ELISA method.Results showed that compared with group A,T-AOC was decreased(P<0.01) and,SOD activity was enhanced in cell culture supernatant(P<0.01) in group B, whereas no significant changes was observed for SOD activity in cell lysate.However,T-AOC of group C was increased significantly(P<0.01) and,SOD activity in cell culture supernatant was much lower than that in group A and group B (P<0.01),but it was higher in cell lysate than that in group A(P<0.05).E.coli treatment(group B) had strong potential to induce APRIL production in Caco-2 cells(P<0.01),but neither IL-8 nor IL-10 production was stimulated.The group C was found to induce production of APRIL(P<0.01),but it was much less than that in group B(P<0.01).In addition,group C showed a significant inhibition to DL-8 production(P<0.01),but in contrast,a powerfull stimultation of IL-10 were determined(P<0.01).Compared with group B,APRIL production was significantly decreased(P<0.01),whereas levels of IL-8(P<0.05) and BL-10(P<0.01) were increased in group D.These results indicated that Lactobacillus rhamnosus could significantly alleviate the inflammatory response of Caco-2 cells against pathogenic bacteria in enterocyte and reinforce cells immune response,which was evidenced by increased antioxidant capacity,increased secretion of pro-inflammatory cytokine APRIL and anti-inflammatory cytokine IL-10,and inhibition of pro-inflammatory cytokine IL-8 production production,and thus offer immuno-protective and immuno-adjuvant effects on Caco-2 cells.更多还原