无菌动物特别是无菌大鼠、小鼠的成功培育技术成为研究微生物与人类生命现象关系的关键技术.严格的无菌动物是指在动物体宿主内外任何部位及生活环境中均检测不出一切生命体.广义的无菌动物还包括清除肠道中大部分菌群的动物,又称为伪无菌动物.目前制作无菌鼠的方法主要有剖腹取胎无菌饲养法制作无菌鼠模型,抗生素鸡尾酒法制作伪无菌鼠模型.本文对无菌鼠模型的制作方法进行总结并对其优缺点进行评价.

Background. Liangxue Tongyu prescription (LTP) is a commonly used formula for acute intracerebral hemorrhage (AICH) in clinical practice that has significant ameliorative effects on neurological deficits and gastrointestinal dysfunction, yet the mechanism remains elusive. The aim of this study was to investigate the pathway by which LTP alleviates brain damage in AICH rats. Methods. The AICH rat models were established by autologous caudal arterial blood injection. The neurological function scores were evaluated before and after treatment. The water content and the volume of Evans blue staining in the brain were measured to reflect the degree of brain damage. RT-PCR was used to detect the inflammatory factors of the brain. Western blotting was used to detect the expression of the tight junction proteins zonula occludens 1 (ZO-1), occludin (OCLN), and claudin (CLDN) in the brain and colon, followed by mucin 2 (MUC2), secretory immunoglobulin A (SIgA), and G protein-coupled receptor 43 (GPR43) in the colon. Flow cytometry was used to detect the ratios of helper T cells 17 (Th17) and regulatory T cells (Treg) in peripheral blood, and the vagus nerve (VN) discharge signals were collected. Results. LTP reduced the brain damage of the AICH rats. Compared with the model group, LTP significantly improved the permeability of the colonic mucosa, promoted the secretion of MUC2, SigA, and GPR43 in the colon, and regulated the immune balance of peripheral T cells. The AICH rats had significantly faster VN discharge rates and lower amplitudes than normal rats, and these abnormalities were corrected in the LTP and probiotics groups. Conclusion. LTP can effectively reduce the degree of brain damage in AICH rats, and the mechanism may be that it can play a neuroprotective role by regulating the function of the intestinal mucosal barrier.

目的:探究凉血通瘀方对脑出血大鼠肠黏膜屏障的组成结构、选择通透性以及神经功能的影响.方法:将30只SD大鼠随机分为正常组、假手术组、模型组、凉血通瘀组和益生菌组.除正常组和假手术组外,其余各组大鼠通过注射泵控制器注血/二次退针法建立脑出血模型.造模成功后,凉血通瘀组给予凉血通瘀方药液灌胃,益生菌组给予双歧杆菌三联活菌制剂灌胃,每日1次,连续3d.治疗结束后,采用ELISA法检测肠黏膜屏障蛋白Occludin、Claudin-1、ZO-1的表达水平及血清中LPS、DAO、DLA的含量.结果:与模型组比较,凉血通瘀组行为学评分显著下降(P<0.01),益生菌组评分改变不明显(P>0.05).与正常组比较,模型组Occludin、Claudin-1、ZO-1蛋白的表达水平显著降低(P<0.01),血清LPS、DAO、DLA的含量显著增加(P<0.01);凉血通瘀组和益生菌组Claudin-1、ZO-1蛋白的表达水平明显降低(P<0.05),血清LPS、DAO、DLA的含量明显增加(P<0.05).与假手术组比较,模型组Claudin-1、ZO-1蛋白的表达水平显著降低(P<0.01),血清LPS、DAO、DLA的含量显著增加(P<0.01),凉血通瘀组ZO-1蛋白的表达水平明显降低(P<0.05),血清LPS、DAO、DLA的含量显著增加(P<0.01);益生菌组血清LPS的含量显著升高(P<0.01).与模型组比较,凉血通瘀组Claudin-1、ZO-1蛋白的表达水平明显上升(P<0.05),益生菌组Occludin、Claudin-1、ZO-1蛋白的表达水平均显著上升(P<0.01).与模型组比较,凉血通瘀组与益生菌组血清LPS、DAO、DLA的含量显著降低(P<0.01).与凉血通瘀组比较,益生菌组血清DLA的含量显著降低(P<0.01).结论:凉血通瘀方与益生菌制剂均可有效提高肠黏膜屏障紧密连接蛋白Occludin、Claudin-1和ZO-1的表达水平,改善肠黏膜通透性,减少LPS、DAO及DLA等有害物质入血,且益生菌制剂稍优于凉血通瘀方.但在改善脑出血后的神经功能缺损症状方面,凉血通瘀方疗效更为确切.

目的:探讨凉血通瘀方对脑出血大鼠脑组织TLR4和NF-κB p65的影响.方法:24只SD健康雄性大鼠随机分成正常组、假手术组、模型组和凉血通瘀组,每组6只.模型组和凉血通瘀组采用注射泵控制器注血/二次退针法制作脑出血模型.造模成功后,凉血通瘀组给予凉血通瘀方水剂(1.0 mL/100 g)灌胃,每日1次;假手术组、模型组每日给予等剂量蒸馏水灌胃.各组灌胃3d后进行大鼠行为学评分;Western Blot法测定Toll样受体4(TLR4)、核因子κB(NF-κB p65)蛋白表达变化;RT-PCR法检测脑组织TLR4 mRNA的表达.结果:与正常组比较,模型组、凉血通瘀组大鼠神经学评分升高(P<0.01);与模型组比较,凉血通瘀组大鼠神经学评分降低(P<0.01).与正常组和假手术组比较,模型组大鼠脑组织TLR4、NF-κB p65蛋白表达皆明显升高(P<0.01),凉血通瘀组NF-κB p65蛋白表达升高(P<0.05);与模型组比较,凉血通瘀组大鼠脑组织TLR4蛋白表达明显下降(P<0.01),NF-κB p65蛋白表达下降(P<0.05).模型组大鼠脑组织TLR4 mRNA的表达明显高于正常组、假手术组及凉血通瘀组(P<0.01);凉血通瘀组与正常组、假手术组未见明显差异.结论:凉血通瘀方可改善脑出血大鼠神经功能损伤,抑制脑组织TLR4和NF-κB p65蛋白表达,凉血通瘀方的作用机制可能与调节TLR4/NF-κB p65信号通路,减轻脑炎性损伤有关.