肠道微生态对三硝基苯磺酸诱导大鼠结肠炎基质金属蛋白酶-3调控作用的研究

目的探讨肠道微生态对三硝基苯磺酸（TNBS）诱导大鼠结肠炎基质金属蛋白酶-3（MMP3）基因表达的调控作用。方法将动物随机分为正常组、模型组、金双歧组3组。采用TNBS法制备大鼠结肠炎模型。造模后金双歧组予以金双歧2g/d经口灌服,1次/d,于第8天评价各组大鼠疾病活动指数（DA1）和组织学损伤评分;ELISA法检测结肠组织TNF-α、IL-10含量;免疫组织化学法检测NF-κB p65的表达;实时荧光定量PCR（RT-PCR）法测定MMP3-mRNA的表达。结果相比模型组,金双歧组大鼠结肠粘膜DAI（5.90±1.67）比（9.20±1.75）]和组织学损伤评分（4.80±1.25）比（7.10±0.99）],TNF-α含量（503.98±126.63）pg/m1比（657.54±149.60）pg/ml],NF-κB p65的表达（3.90±2.02）比（6.10±1.79）],MMP3-mPNA的表达（3.56±2.13）比（16.53±13.80）]均降低（P均〈0.01或P〈0.05）,而IL-10含量（95.57±27.71）pg/ml比（42.92±23.74）pg/ml]增高（P〈0.01）;NF-κBp65的表达与MMP3-mRNA表达呈显著正相关（相关系数γ=0.669,P〈0.05）。结论金双歧对TNBS诱导大鼠实验性结肠炎有明显的抗炎效果,其作用机制可能是通过下调MMP3-mRNA的表达、抑制NF-κB活性、平衡细胞因子网络等实现的。

Study on the intestinal microecology in contols matrix metalloproteinase-3 expression in colonic mucosa of rat with trinitrobenzenesulfonic acid induced colitis

Objective To explore the regulation of the intestinal microecology on the gene expression of Matrix Metalloproteinase-3（MMP3） in colonic mucosa of rat with 2,4,6-trinitrobenzene sulfonic acid（TNBS）induced colitis.Methods The animals were randomized into 3 groups;golden bifid group,model group and normal group.Rat colitis models were established by TNBS enema.The golden bifid group rats were drenched with golden bifid at the dosage of 2g/d,once a day.On the eighth day,the disease acticity index（DAI）and the score of the tissue damage of colonic mucosa was calculated;the level of tumor necrosis factor-α（TNF-α）and interleukin-10（IL-10）was defected by enzyme linked immune sorbent assay（ELISA）;the expression of NF-κB p65 in colonic tissue was observed by immunohistochemistry：the expression of MMP3-mRNA was detected by realtime fluorescent quantitation polymerase chain reaction（RT-PCR）.Results Compared with model grope,the golden bifid group rats DAI（5.90±1.67）vs（9.20±1.75）]and the score of the tissue damages（4.80±1.25）vs（7.10±0.99）],the levels of TNF-α（503.98±126.63）pg/ml vs（657.54±149.60）pg/ml],the expression of NF-κBp65（3.90±2.02）vs（6.10±1.79）]and the expression of MMP3-mRNA（3.56±2.13）vs（16.53±13.80）]were all significantly decreased（P0.01 or P0.05）,but that of IL-10（95.57±27.71）pg/ml vs（42.92±23.74）pg/ml]were significantly increased（P0.01）;the expression of MMP3-mRNA has a significant positive correlation with the expression of NF-KB p65（γ=0.669,P0.05）.Conclusion Golden bifid has an exact anti-inflammatory effects on TNBS induced rat colitis.The mechanism is possibly related to down-regulated the expression of MMP3-mRNA,inhibit the activation of NF-κB and regulate network balance of cytokines and et al.