免疫球蛋白A肾病大鼠模型建立的方法学研究  

Methodological study of establishing of immunoglobulin A nephropathy rat model

作　　者：马思佳 杨斌[2] 赵明明[1] 常美莹 潘知玉 范娇 张紫嫣 薛舜暄 张昱[1] MA Si-jia;YANG Bin;ZHAO Ming-ming;CHANG Mei-ying;PAN Zhi-yu;FAN Jiao;ZHANG Zi-yan;XUE Shun-xuan;ZHANG Yu(Department of Nephrology,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;Department of Pathology,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China)

机构地区：[1]中国中医科学院西苑医院肾病科,北京100091 [2]中国中医科学院西苑医院病理科,北京100091

出　　处：《中国临床药理学杂志》2022年第17期2049-2054,共6页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金面上基金资助项目(81873300);首都卫生发展科研专项基金资助项目(2018-2-4173);首都临床诊疗技术研究及示范应用基金资助项目(Z191100006619063)。

摘　　要：目的 分2个时间点评价不同剂量牛血清白蛋白(bovine serum albumin,BSA)灌胃联合尾静脉注射脂多糖(lipopolysaccharide,LPS),皮下注射四氯化碳(carbon tetrachloride,CCl4)方法建立免疫球蛋白A(immunoglobulin A,IgA)肾病大鼠模型的效果。方法 将SPF级健康雄性SD大鼠50只随机分为低剂量纯化水组(A组)、低剂量酸化水组(B组)、高剂量纯化水组(C组)、高剂量纯化水连续尾静脉组(D组)和空白组(E组),每组10只。A组给予400 mg·kg^(-1)纯化水配制BSA 500 mL,隔天灌胃1次,持续9周,蓖麻油0.3 mL+CCl_(4) 0.1 mL,每周皮下注射1次,共9周,于第6,8周尾静脉注射LPS 0.05 mg。B组给予400 mg·kg^(-1)酸化水配制BSA 500 mL,隔天灌胃1次,其余操作同A组。C、D 2组均给予600 mg·kg^(-1)纯化水配制BSA 500 mL,隔天灌胃1次,皮下操作同前,时间均延长至12周;分别于第8周和第10周对C组大鼠尾静脉注射LPS 0.05 mg。从第8周至第12周,每周对D组大鼠尾静脉注射LPS 0.05 mg。E组给予0.9%NaCl,操作同A组,持续12周。分别于第6,8,10和12周用考马斯亮蓝法测定大鼠24 h尿蛋白定量,用苏木精-伊红染色法观察大鼠病理组织,用免疫荧光法测定大鼠IgA沉积强度。结果 在第12周时,A、B、C、D、E组的24 h尿蛋白定量达到最高值,分别为(12.00±1.79),(12.71±3.40),(15.17±3.31),(13.43±2.23)和(4.05±0.82)mg·d^(-1),E组与其他4组相比,差异均有统计学意义(均P<0.05)。与第9周末相比,在第12周末时各模型组大鼠的肾病理病变更严重,IgA荧光沉积更显著,其中B、C组的IgA免疫荧光计数与E组相比,差异均有统计学意义(均P<0.01),且C组的IgA免疫荧光强度较B组更强(P<0.05)。结论 以纯化水配制高剂量BSA(600 mg·kg^(-1))+LPS+CCl_(4),操作12周可建立肾组织病变更显著、更典型的IgA肾病大鼠模型。Objective To evaluate the effects of different doses of bovine serum albumin(BSA) combined with lipopolysaccharide(LPS) and carbon tetrachloride(CCl_(4)) on the establishment of immunoglobulin A(IgA) nephropathy rat model at two time points.Methods Fifty SPF healthy male SD rats were randomly divided into low-dose purified water group A,low-dose acidified water group B,high-dose purified water group C,high-dose purified water continuous caudal vein group D and normal group E,with 10 rats in each group.The concentration of BSA in group A and group B was 400 mg·kg^(-1),and group B was treated with acidified water.The rats in the two groups were subcutaneously injected with castor oil 0.3 mL + CCl_(4) 0.1 mL per week for 9 weeks,and LPS was injected with 0.05 mg in caudal vein at week 6 and 8.Group C and group D were treated with 600 mg · kg-1purified water configuration with BSA 500 m L,gavaged once the next day,the subcutaneous operation was the same as before and was extended to 12 weeks.Group C was injected with LPS 0.05 mg in caudal vein at week 8 and 10.LPS0.05 mg was administered weekly in the tail vein in group D rats from weeks 8 to 12.The group E was treated with the same amount of normal saline as group A for 12 weeks.At 6,8,10 and 12 weeks,24 h urinary protein was determined by Coomassie bright blue method,pathological tissues were observed by hematoxylin-eosin staining,and Ig A deposition intensity was determined by immunofluorescence method.Results At 12 weeks,24 h urinary protein quantification in groups A,B,C,D and E reached the highest value( 12.00 ± 1.79),( 12.71 ± 3.40),( 15.17 ± 3.31),( 13.43 ±2.23) and( 4.05 ± 0.82) mg·d^(-1),respectively,and the differences were statistically significant compared with group E( all P < 0.05).Compared with 9 weeks,renal pathological changes were more serious and Ig A fluorescence deposition was more significant in each model group at 12 weeks.The immunofluorescence counts of group B and C were significantly different from those of group E( all P < 0.01),a

关 键 词：免疫球蛋白A肾病 大鼠 模型 牛血清白蛋白 脂多糖 四氯化碳 

分 类 号：R97[医药卫生—药品]