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目的探讨和分析实验性结肠炎大鼠结肠Toll样受体2(TLR2)和Toll样受体4(TLR4)的表达及意义,并分析益生菌治疗的作用。方法30只雄性Wistar大鼠均分为正常对照组(NC组)、模型对照组(UC组)和益生菌治疗组(PC组)。UC和PC组建立2,4,6-三硝基苯磺酸(TNBS)实验性结肠炎大鼠模型。PC组大鼠给予双歧三联活菌悬液(2.2×10~9 CFU/只)治疗,1次/d,共4周。光镜下观察肠黏膜炎性反应片评分。Western印迹法和实时荧光定量PCR法检测大鼠结肠TLR2和TLR4蛋白及mRNA表达,分析其变化及益生菌治疗的作用。结果NC、PC和UC组炎性反应评分分别为4.35±0.88、10.25±1.36和7.94±0.85。PC组炎性反应评分较UC组明显改善(P<0.05),但高于NC组水平(P<0.01)。NC、UC、PC组TLR2和TLR4蛋白表达水平分别为10.26±4.24、47.20±4.62、36.64±3.67和8.16±4.50、25.84±4.46、19.71±3.28。二者在UC组的表达明显高于NC组(P<0.01)和PC组(P<0.05),PC组高于NC组(P<0.01)。NC、UC、PC组TLR2和TLR4 mRNA表达水平分别为240±140、10570±2690、6640±1420和210±110、8580±2710、5290±1540。二者在UC组的表达明显高于PC组(P<0.05)和NC组(P<0.01),PC组高于NC组(P<0.01)。结论实验性结肠炎大鼠结肠TLR2和TLR4表达明显增高,提示肠腔内抗原摄取增多。双歧三联活菌可明显降低二者的表达水平,提示该益生菌可能通过降低TLR2和TLR4表达减少肠腔抗原的摄取。
Objective To investigate and analyze the expression and significance of toll - like receptor 2 (TLR2) and toll - like receptor 4 (TLR4) in experimental colitis in rats and to analyze the effect of probiotics treatment. Methods Thirty male Wistar rats were divided into normal control group (NC group), model control group (UC group) and probiotics treatment group (PC group). A rat model of experimental colitis of 2,4,6-trinitrobenzene sulfonic acid (TNBS) was established in UC and PC groups. The rats in PC group were treated with 2.2 × 10 ~ 9 CFU / suspension of Bifidobacterium triplex, once a day for 4 weeks. Light microscopic observation of intestinal mucosal inflammatory response score. The expression of TLR2 and TLR4 protein and mRNA in the colon of rats were detected by Western blotting and real-time fluorescence quantitative PCR, and their changes and the effect of probiotics treatment were analyzed. Results The inflammatory response scores in NC, PC and UC groups were 4.35 ± 0.88, 10.25 ± 1.36 and 7.94 ± 0.85, respectively. PC group inflammatory response score significantly improved compared with UC group (P <0.05), but higher than the NC group (P <0.01). The protein expression levels of TLR2 and TLR4 in NC, UC and PC groups were 10.26 ± 4.24, 47.20 ± 4.62, 36.64 ± 3.67 and 8.16 ± 4.50, 25.84 ± 4.46 and 19.71 ± 3.28, respectively. The expression of both in UC group was significantly higher than NC group (P <0.01) and PC group (P <0.05), PC group was higher than NC group (P <0.01). The mRNA expressions of TLR2 and TLR4 in NC, UC and PC groups were 240 ± 140,10570 ± 2690,6640 ± 1420 and 210 ± 110,8580 ± 2710,5290 ± 1540, respectively. The expression of both in UC group was significantly higher than that in PC group (P <0.05) and NC group (P <0.01), and in PC group was higher than that in NC group (P <0.01). Conclusion The expressions of TLR2 and TLR4 in the colon of experimental colitis rats were significantly increased, suggesting that the intra-intestinal antigen uptake increased. Bifidobacterium triple viable bacteria can significantly reduce the expression of both, suggesting that the probiotics may reduce the intestinal cavity antigen by TLR2 and TLR4 expression.