论文部分内容阅读
目的研究基因转染上调Smad7表达对大鼠腹膜纤维化模型腹膜炎症细胞浸润及促炎症因子表达的影响。方法 48只SD大鼠随机分为(1)正常对照组(n=12);(2)模型组(n= 12):每日给予腹腔注射4．25％葡萄糖腹膜透析液(100 ml／kg),同时于第8、10、12、22、24、26天腹腔注射脂多糖(LPS,0．6 mg／kg);(3)空白载体对照组(n=12):仅转入不含Smad7的Tet-on／ vector空白载体:(4)Smad7基因转染组(n=12):在造模后第0、14天分别转染Smad7。第28天时杀检,分别应用免疫荧光染色及RT-PCR检测脏层腹膜泛白细胞标志性抗原(OX-1)、单核巨噬细胞抗原(ED-1)、白介素1(IL-1)、肿瘤坏死因子α(TNF-α)蛋白和mRNA的表达水平。Smad7 基因转染采用超声介导的微泡基因转染技术。结果与模型组及空白载体转染组比较,Smad7转基因组脏层腹膜OX-1、ED-1阳性细胞以及IL-1、TNF-α表达水平有明显的减少,但均高于正常对照组。结论高糖腹膜透析液联合LPS可刺激腹膜炎症细胞的浸润及腹膜间皮细胞IL-1、 TNF-α表达上调。基因转染上调Smad7表达可明显抑制大鼠腹膜纤维化模型腹膜炎症细胞浸润及促炎症因子的表达上调。
Objective To investigate the effects of up-regulation of Smad7 gene transfection on peritoneal inflammatory cell infiltration and proinflammatory cytokine expression in rat peritoneal fibrosis model. Methods 48 SD rats were randomly divided into normal control group (n = 12), model group (n = 12): intraperitoneal injection of 4.25% glucose peritoneal dialysis solution (100 ml / kg ), While lipopolysaccharide (LPS, 0.6 mg / kg) was injected intraperitoneally on the 8th, 10th, 12th, 22th, 24th and 26th day; (3) Blank vector control group Tet-on / vector blank vector of Smad7: (4) Smad7 gene transfection group (n = 12): Smad7 was transfected on the 0th and the 14th day after model establishment. On the 28th day, the mice were sacrificed and the visceral peritoneal leukocyte antigen (OX-1), monocyte-macrophage antigen (ED-1), interleukin-1 (IL- Tumor necrosis factor alpha (TNF-alpha) protein and mRNA expression levels. Smad7 gene transfection using ultrasound-mediated gene transfer of microbubbles. Results The expression of OX-1, ED-1 positive cells, IL-1 and TNF-α in the peritoneum of Smad7 transgenic group were significantly decreased compared with the model group and blank vector transfection group, but both were higher than the normal control group. Conclusion High glucose and peritoneal dialysis solution combined with LPS can stimulate peritoneal inflammatory cells infiltration and peritoneal mesothelial cells IL-1 and TNF-α upregulated. Up-regulation of Smad7 gene transfection can significantly inhibit the peritoneal fibrosis rat model of peritoneal inflammatory cell infiltration and up-regulation of pro-inflammatory cytokines.