多因素所致IgA肾病模型大鼠血清炎症因子水平和肾组织免疫相关蛋白表达的变化

Changes in expressions of serum inflammation factor levels and renal tissue immune-associated proteins in multiple factors-caused IgA nephropathymodel rats

目的通过多因素所致IgA肾病(IgA nephropathy,IgAN)大鼠模型,观察大鼠血清炎症因子以及肾组织T细胞免疫球蛋白黏蛋白-1(T cell immune globulin sticky protein 1,TIM-1)和转化生长因子-β1(transforming growth factor-β1,TGF-β1)的表达变化,探讨IgAN的炎症免疫机制。方法取20只SPF级雄性SD大鼠,采用随机数字表分为模型组和空白对照组。模型组予以牛血清白蛋白溶液(400 mg/kg,1 mL/100g)隔日灌胃,持续6周;皮下注射蓖麻油0.5 mL+四氯化碳溶液0.1 mL,每周1次,持续9周;并给予脂多糖(LPS)溶液(0.05 mg/只),分别于第6、8周尾静脉注射。空白对照组予以等体积纯化水隔日灌胃,相同时间点皮下注射0.9%氯化钠溶液0.4 mL/只和尾静脉注射0.9%氯化钠溶液0.2 mL/只,造模时间为10周。造模后,检测两组大鼠尿微量白蛋白,24 h尿蛋白定量及肝、肾功能指标;采用ELSIA法检测大鼠血清中IgA、干扰素-γ(IFN-γ)、白细胞介素-4(IL-4)、白细胞介素-6(IL-6)的含量及肾脏组织中TGF-β1和TIM-1的表达。结果造模后第8周,大鼠出现不同程度的毛色灰暗、精神萎靡、便溏等症状。与空白对照组比较,模型组大鼠尿微量白蛋白和24 h尿蛋白定量显著升高(P<0.05);血清IgA含量和肾组织的IgA荧光表达均显著升高(P<0.05)。与空白对照组比较,模型组大鼠的血清IL-4、IL-6水平显著增高(P<0.05),肾组织的TGF-β1和TIM-1表达显著增强(P<0.05)。结论多因素所致IgAN模型和临床IgA患者体征相似,其发病机制和炎症因子释放与TGF-β1及TIM-1的表达密切相关。

Objective To observe the changes in expressions of serum inflammation factors,kidney tissues T cell immune globulin sticky protein 1(TIM-1)and transforming growth factor-beta 1(TGF-beta 1)through multiple factors-caused IgA nephropathy model rats,and to investigate the inflammatory immune mechanism of IgA nephropathy.Methods Twenty healthy male Sprague-Dauley rats were randomly divided(Random number table)into 2 groups with a random digit table:control group(n=10),and model group(n=10).The rats in the model group were administered gastrically with bovine serum albumin solution(400 mg/kg,1 mL/100g)every other day for 6 weeks,and injected subcutaneously with 0.5 mL castor oil and 0.1mL carbon tetrachloride solution once a week for 9 weeks,and injected intravenously through the tail vein with lipopolysaccharide(LPS)solution(0.05 mg/rat)in week 6 and week 8.The rats in the control group were administered gastrically with the same amount of distilled water,and injected subcutaneously with 0.9%sodium chloride solution at a dose of 0.4 mL/rat at the same time points,and injected intravenously through the tail vein with 0.9%sodium chloride solution at a dose of 0.2 mL/rat.The modeling time was 10 weeks.After modeling,urinary microalbumin,24h urine protein and liver and kidney function indices were determined for each group.ELSIA method was used to detect contents of IgA,IFN-γ,IL-4,and IL-6 in the serum,and expressions of TGF-β1 and TIM-1 in rat kidney tissues.Results In the model group,in week 8 after modeling the rats developed less shiny fur color with different severities,listlessness,loose stools.Compared to the rats in the control group,the rats in the model group had significantly increased urinary microalbumin and 24h urine protein(P<0.05),and obviously increased IgA content in the serum and fluorescent expressions of IgA in renal tissues(P<0.05).Compared to the rats in the control group,the rats in the model group showed notably increased serum IL-4 and IL-6 levels(P<0.05),significantly enhanced expressions of GF-β1 and TIM-1 in renal tissues(P<0.05).Conclusion:The multiple factors-caused IgAN model rats have the signs similar to clinical patients with IgAN.Its pathogenesis is closely associated with release of inflammatory factors and expression of TGF-beta 1 and TIM-1.