培菲康对实验性溃疡性结肠炎大鼠TNF-α、IL-10水平的影响

The effect of live combined bifidobacterium,lactobacillus and enterococcus on TNF-αand IL-10 level in experimental ulcerative colitis of rats

目的阐明培菲康对三硝基苯磺酸钠(TNBS)诱导的大鼠溃疡性结肠炎(UC)TNF-α、IL-10水平的影响,寻求UC治疗的有效新途径。方法成年雌性SD大鼠50只,随机分成5组(n=10):正常对照组(G1)、模型对照组(G2)、培菲康治疗组(G3)、奥沙拉嗪治疗组(G4)、培菲康和奥沙拉嗪联合治疗组(G5),经不同处理和治疗后,通过免疫组化染色观测各组大鼠结肠组织中TNF-α(两步法)、IL-10(S-P法)表达情况,通过实时荧光定量PCR分析结肠组织中TNF-α、IL-10mRNA的表达水平。结果G1组各指标显著低于G2组(P<0.001),肠组织结构正常;与G2组相比,G3、G4、G5组TNF-α、IL-10细胞阳性率和TNF-α、IL-10mRNA的表达水平显著降低(P<0.001);与G3或G4组相比,G5组TNF-α、IL-10细胞阳性率和TNF-α、IL-10mRNA的表达水平也显著降低(P<0.001);G3或G4组之间,TNF-α、IL-10细胞阳性率及TNF-αmRNA水平无差别(P>0.05),而G4组IL-10mRNA水平明显低于G3组(P<0.001)。结论TNF-α、IL-10在溃疡性结肠炎(UC)的发生、发展过程中起重要作用,培菲康能明显降低该过程中肠组织TNF-α、IL-10的表达水平,可能通过调控这两个细胞因子的表达而发挥疗效。

Objective To investigate the effect of live combined bifidobacterium,lactobacillus and enterococcus capsules on ulcerative colitis（UC）in rats induced by trinitrobenzene sulfonic acid（TNBS）,so as to explore a new and effective therapy for UC.Method Fifty female adult Sprague-Dawley rats were randomly divided into 5 groups（n=10）,normal control group（G1）,untreated TNBS-induced colitis group（G2）,TNBS-induced colitis treated with live combined bifidobacterium,lactobacillus and enterococcus group（G3）,TNBS-induced colitis treated with olsalazine group（G4）and TNBS-induced colitis treated with both live combined bifidobacterium,lactobacillus and enterococcus and olsalazine in the same dose and duration group（G5）.Each group received different treatment as described above,and tumor necrosis factor-alpha（TNF-α）,interleukin-10（IL-10）in colonic mucosa were observed through immunohistochemical stain（Two-step method for TNF-a,Streptavidin-Peroxidase method for IL-10）and analysized by multimedia pathological analysis system.Colonic mRNA levels of TNF-α and IL-10 were determined using real-time quantitative reverse transcriptase polymerase chain reaction（RT-PCR）.Results Parameters of G1 were significantly lower than those of G2（P〈0.001）;G3,G4 and G5 groups showed greater cellular positive rates of TNF-αand IL-10 and mRNA levels of TNF-αand IL-10（P〈0.001）when compared to G2;G5 showed more notable colonic TNF-αand IL-10 cellular positive rates and mRNA levels in comparison woth either G3 or G4（P〈0.001）;Colonic cellular positive rates of TNF-αand IL-10 and TNF-αmRNA level were not significantly different between G3 and G4 group（P〉0.05）,however,mRNA level of IL-10 in G4 was much lower than that in G3（P〈0.001）.Conclusion TNF-αand IL-10 are both playing very important roles in the pathogenesis and development of ulcerative colitis.live combined bifidobacterium,lactobacillus and enterococcus can effectively downregulate the expressions of TNF-αand IL-10 in rats induced by TNBS,and its potential therapeutic effect may be modulating the expressions of these two cytokines.