CAJ | 학술논문

目的：验证功能蛋白烯醇化酶（enolase）和Tuf蛋白是否为长双歧杆菌（Bifidobacterium longum）NCC2705的黏附因子。方法将长双歧杆菌NCC2705与Caco-2细胞共培养，显微镜观察加入纯化蛋白烯醇化酶、Tuf蛋白前后细胞黏附细菌数的变化并进行统计学分析。将Caco-2细胞、长双歧杆菌NCC2705、致病菌（弗氏志贺菌或伤寒沙门菌）共培养，检测加入纯化蛋白烯醇化酶、Tuf蛋白前后上清液中乳酸脱氢酶（ LDH）和肿瘤坏死因子-α（ TNF-α）的水平。结果加入纯化蛋白烯醇化酶、Tuf蛋白之后，Caco-2细胞黏附细菌数显著降低（P＜0．05）。 Caco-2细胞与致病菌共培养时，LDH和TNF-α的水平显著增高（P＜0．05）；Caco-2细胞与长双歧杆菌NCC2705和致病菌共培养时，LDH和TNF-α的水平显著降低（P＜0．05），然而加入纯化蛋白烯醇化酶和Tuf蛋白后，LDH和TNF-α的水平又出现显著增高（P＜0．05）。结论长双歧杆菌NCC2705可竞争性抑制致病菌对肠上皮细胞的黏附，而烯醇化酶和Tuf蛋白可竞争性抑制长双歧杆菌对肠上皮细胞黏附作用，是双歧杆菌的黏附因子。
목적：험증공능단백희순화매（enolase）화Tuf단백시부위장쌍기간균（Bifidobacterium longum）NCC2705적점부인자。방법장장쌍기간균NCC2705여Caco-2세포공배양，현미경관찰가입순화단백희순화매、Tuf단백전후세포점부세균수적변화병진행통계학분석。장Caco-2세포、장쌍기간균NCC2705、치병균（불씨지하균혹상한사문균）공배양，검측가입순화단백희순화매、Tuf단백전후상청액중유산탈경매（ LDH）화종류배사인자-α（ TNF-α）적수평。결과가입순화단백희순화매、Tuf단백지후，Caco-2세포점부세균수현저강저（P＜0．05）。 Caco-2세포여치병균공배양시，LDH화TNF-α적수평현저증고（P＜0．05）；Caco-2세포여장쌍기간균NCC2705화치병균공배양시，LDH화TNF-α적수평현저강저（P＜0．05），연이가입순화단백희순화매화Tuf단백후，LDH화TNF-α적수평우출현현저증고（P＜0．05）。결론장쌍기간균NCC2705가경쟁성억제치병균대장상피세포적점부，이희순화매화Tuf단백가경쟁성억제장쌍기간균대장상피세포점부작용，시쌍기간균적점부인자。
Objective To demonstrate that enolase and Tuf are direct adhesions of Bifidobacterium longum( B.longum) NCC2705.Methods B.lougum NCC2705 was co-cultured with Caco-2.The change of adhesion was tested by microscopy after the purified proteins(GST-Eno and GST-Tuf) were added into the co-culture, and then statistical analysis was conduc-ted.Caco-2 cells were incubated with B.lougum NCC2705 and pathogenic bacteria (Shigella flexneri 2a or Salmonella). The level ofcytokine LDH and TNF-αfor supernatant was tested after the purified proteins ( GST-Eno and GST-Tuf) were added.Results The number of bacteria adherent to Caco-2 was reduced significantly ( P <0.05 ) after the purified proteins ( GST-Eno/GST-Tuf) were added .While Caco-2 was co-cultured with pathogens , the level of LDH and TNF-αwas significantly increased (P<0.05).When Caco-2 was incubated with B.lougum NCC2705 and pathogenic bacteria , the level of LDH and TNF-αwas significantly reduced (P<0.05), but the level of LDH and TNF-αincreased once more (P<0.05) after the purified protein (GST-Eno/GST-Tuf) was added.Conclusion B.longum NCC2705 can competitive-ly inhibit production of pathogenic bacteria in intestinal epithelial cell adhesion , while the enolization enzyme and Tuf proteins can competitively inhibit long bifidobacteria in intestinal epithelial cell adhesion effect .They are adhesion factors of bifidobacteria .