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目的探讨纳米四氧化三铁双歧杆菌脂磷壁酸(Fe3O4-LTA)复合物缓解溃疡性结肠炎(ulcerative coli-tis,UC)的情况。方法 100只Babl/c小鼠随机分为5组,即阴性对照组(N)、溃疡性结肠炎阳性对照组(UC)、0.1 mg/mL纳米Fe3O4-LTA缓解组(L)、0.5 mg/mL纳米Fe3O4-LTA缓解组(M)、1 mg/mL纳米Fe3O4-LTA缓解组(H)。N组不施加作用,UC组、缓解组(L组、M组、H组)用2%葡聚糖硫酸钠(dextran sodium sulphate,DSS)自由饮用2周,2周后UC组不施加作用,L、M、H组分别采用0.1 mg/mL、0.5 mg/mL、1 mg/mL灌肠1 mL,每天1次,连续3周。观察临床体征和检测WBC、CRP、PCT、IL-18、IL-10。灌肠3周后处死小鼠,分离结肠黏膜上皮细胞,用RT-PCR法检测TLR2、TLR4的表达,用Western blot和免疫组织化学检测NF-κBp65的表达,组织病理切片评估缓解效果。结果纳米Fe3O4-LTA缓解1周后小鼠精神状态明显恢复良好,活动及进食增加,毛色有光泽。缓解组比UC组体重增幅大(P<0.05);血液检测发现缓解组炎性指标不同程度下降,其中H组与其他组相比差异具有统计学意义(P<0.05);RT-PCR检测显示:UC组与缓解组结肠黏膜上皮细胞内TLR2的表达要高于N组(P<0.05),UC组TLR4的表达要高于N组和缓解组(P<0.05)。NF-κBp65的Western blot和免疫组织化学染色显示UC组与缓解组的表达要高于N组(P<0.05);病理组织切片和临床体征显示总缓解率达83.33%。结论纳米Fe3O4-LTA复合物具有较好缓解UC的能力。
Objective To investigate the alleviation of ulcerative colitis (UC) by nano-Fe3O4-LTA complex. Methods 100 Babl / c mice were randomly divided into 5 groups: negative control group (N), ulcerative colitis positive control group (UC), 0.1 mg / mL Fe3O4-LTA remission group mL nano-Fe3O4-LTA remission group (M) and 1 mg / mL nano-Fe3O4-LTA remission group (H). The rats in the UC group and the remission group (L group, M group and H group) were free to drink 2% dextran sodium sulphate (DSS) for 2 weeks. After 2 weeks, the rats in UC group did not exert any action, L, M, H group were 0.1 mg / mL, 0.5 mg / mL, 1 mg / mL enema 1 mL once a day for 3 weeks. Clinical signs and detection of WBC, CRP, PCT, IL-18, IL-10 were observed. After 3 weeks, the mice were killed and colon mucosal epithelial cells were isolated. The expression of TLR2 and TLR4 was detected by RT-PCR. The expression of NF-κBp65 was detected by Western blot and immunohistochemistry. Results After 1-week remission of Fe3O4-LTA, the mental state of mice was recovered well and their activities and food intake increased. The body weight of rats in the remission group was significantly higher than that of the UC group (P <0.05). The blood test showed that the inflammatory index of the remission group decreased to different extents. The difference between the H group and other groups was statistically significant (P <0.05) : The expression of TLR2 in UC and remission group was higher than that in N group (P <0.05). The expression of TLR4 in UC group was higher than that in N group and remission group (P <0.05). Western blot and immunohistochemical staining of NF-κBp65 showed that the expression of NF-κBp65 was higher in group UC and remission than that in group N (P <0.05). Pathological sections and clinical signs showed a total remission rate of 83.33%. Conclusion Fe3O4-LTA nanocomposites have better ability to relieve UC.