益肾通络方对膜性肾病SD大鼠肾组织纤溶酶原激活物抑制因子-1及生化、病理指标的影响

目的观察益肾通络方对膜性肾病(MN)SD大鼠肾组织纤溶酶原激活物抑制因子-1(PAI-1)及生化、病理指标的影响。方法将40只SD大鼠随机分为正常对照组、模型组、西药组及中药组4组,每组各10只。模型组、西药组及中药组大鼠予阳离子化牛血清白蛋白(C-BSA)2.5 mg(溶于1 mL pH值7.4磷酸盐缓冲液)尾静脉注射,对照组予等容积0.9%氯化钠注射液尾静脉注射,每周二、五、日各1次,连续4周。采用全自动生化分析仪测定各组大鼠血清白蛋白(Alb)、总蛋白(TP)、甘油三酯(TG)及总胆固醇(TC);采用双缩脲法测定24 h尿蛋白定量;采用间接免疫荧光法检测免疫球蛋白G(IgG)和补体3(C3),采用透射电镜观察肾小球基底膜及足突变化;采用链球菌抗生物素蛋白-生物素-过氧化物酶复合物(SABC)法进行PAI-1免疫组化检测。结果模型组、西药组及中药组SD大鼠血清Alb、TP均低于正常对照组(P<0.05),TG、TC及24 h尿蛋白定量均高于正常对照组(P<0.05);西药组、中药组血清Alb、TP均高于模型组(P<0.05),TG、TC及24 h尿蛋白定量均低于模型组(P<0.05);中药组与西药组血清Alb、TP、TG、TC及24 h尿蛋白定量比较差异无统计学意义(P>0.05)。模型组、西药组及中药组大鼠肾组织均有不同程度的IgG及C3呈细颗粒状沿毛细血管袢及部分系膜区沉积,与模型组比较,中药组和西药组沉积显著减少。模型组大鼠肾小球基底膜增厚,西药组及中药组与模型组比较明显减轻。PAI-1主要表达于肾小球和肾小管上皮细胞内,在正常组中有微量表达。模型组、中药组及西药组大鼠肾组织中PAI-1的表达较正常对照组增强(P<0.05),而中药组及西药组较模型组明显减轻(P<0.05),中药组与西药组比较差异无统计学意义(P>0.05)。结论益肾通络方可显著降低MN SD大鼠尿蛋白,提高血浆蛋白,改善血脂代谢,抑制PAI-1在肾组织中的表达,减轻足突融合,促进受损的肾小球基底膜修复,从而减轻肾脏损伤,延缓肾脏慢性病理进展。

Effects of Yishentongluo decoction on the expression of PAl - 1, biochemical and pathological indicators inrat with experimental membranous nephropathy

Objective To observe the Effects of Yishentongluo Decoction on the expression of PAl - 1 on experimental membranous nephropathy. Methods 40 SD rats were randomly divided into four groups; control group, model group, western medicine group and Chinese herb group （ n = 10, in each group）. Model group, west- em medicine group and Chinese herb group received tail vein injection of 2.5 mg of cationic bovine serum albumin （ C - BSA） （ dissolved in 1 mL pH value of 7.4 phosphate buffer） , 3 times a week for 4 weeks. After the modeling, the normal group and model group were fed by saline. Western medicine group received benazepril tablets dissolved in 5ml normal saline at 10 mg/kg daily. Chinese herb group received Yishentongluo decoction 5 mL, once a day for 4 weeks. Mouse serum albumin （Alb） , total protein （TP）, triglycerides （TG） and total cholesterol （TC） were detec-ted using automatic biochemical analyzer; the determination of 24 h urinary protein was completed by biuret method for; immunoglobulin G （ IgG ） and complement 3 （ C3 ） were measured by indirectimmunofluorescence detection, the glo- merular basement membrane and foot process changes were observed using transmission electron microscopy, PAI- Iimmunohistochemistry was completed by the use of streptococcus avidin -biotin -peroxidase complex （SABC） meth- od. Results TP and Alb of the blood serum in model group, western medicine group and Chinese herb group were significantly decreased as compared with those in control group （P 〈 0.05）. The levels of TG, TC and 24 h urinary protein quantitative in model group, western medicine group and Chinese herb group were higher than those in control group （P 〈 0.05）. There was no significant difference between western medicine group and Chinese herb group on Alb, TP, TG, TC and 24 h urinary protein quantitative （ P 〉 0.05 ）. IgG and C3 in model group, western medicine and Chinese herb group showed granular climbing along the capillary and part of deposition. Deposition in westernmedicine group and Chinese herb group was significantly decreased as compared with that in model group （ P 〈 0. 05 ）. The thickness of the glomerular basement membrane in model group was increased. The thickness of the glomer- ular basement membrane in western medicine group and Chinese herb group was decreased as compared with that in model group （ P 〈 0.05 ）. PAI- 1 was mainly expressed in glomerular and tubular epithelial cells, trace expression in the normal group. The expression of PAI- 1 in model group, western medicine group and Chinese herb group was increased as compared with that in control group （P 〈 0.05）. The expression of PAI- 1 in western medicine group and Chinese herb group was decreased as compared with that in model group （P 〈 0.05 ）. There was no marketly difference between the western medicine group and Chinese herb group （ P 〉 0.05 ）. Conclusion Yishentongluo decoction can reduce proteinuria, increase plasma protein, improve blood rheology to protect renal function. Yishent- ongluo Decoction can depress PAl - 1 expression in nephridial tissue.