摘要
【目的】观察肾炎消白颗粒对肾小球肾炎蛋白尿模型大鼠肾脏组织中Nephrin mRNA动态表达的影响,探讨其作用机理。【方法】将100只Wistar雄性大鼠随机分为空白组、模型组、洛汀新组、中药低剂量组、中药高剂量组,每组20只。采用一次性尾静脉注射阿霉素复制肾小球肾炎蛋白尿大鼠模型。洛汀新组大鼠给予洛汀新(剂量为0.90 mg·kg-1·d-1)灌胃,中药高、低剂量组给予肾炎消白颗粒(剂量分别为3.60、1.80 g·kg-1·d-1)灌胃,每天1次,1周连续给药6次,共给药7周。每周测量1次大鼠体质量,于第3、5、7周分别采用苏木素—伊红(HE)染色法观察各组大鼠肾组织病理学改变,采用实时荧光定量聚合酶链反应(RT-q PCR)法检测各组大鼠肾组织Nephrin mRNA表达。【结果】(1)体质量:与空白组比较,模型组大鼠体质量出现明显的异常增加,尤以造模3周后更明显(P<0.05);与模型组比较,洛汀新组大鼠各时间点体质量的异常增加均得到有效控制(P<0.05),中药高剂量组在造模3周后也得到有效控制(P<0.05),且造模3周后中药高剂量组的控制效果优于洛汀新组(P<0.05)。(2)尿白蛋白/尿肌酐比值:与空白组比较,模型组大鼠各时间点尿白蛋白/尿肌酐比值显著升高(P<0.01);与模型组比较,洛汀新组大鼠各时间点尿白蛋白/尿肌酐比值显著降低,中药高剂量组大鼠在第3、4、5、6、7周尿白蛋白/尿肌酐比值显著降低(P<0.05);与洛汀新组比较,中药高剂量组大鼠在第5、6、7周尿白蛋白/尿肌酐比值降低(P<0.05)。(3)Nephrin mRNA:与空白组比较,模型组大鼠第3周肾组织Nephrin mRNA表达显著升高,于第5、7周显著降低(P<0.05);与模型组比较,中药高剂量组大鼠各时间点肾组织Nephrin mRNA表达显著升高(P<0.05);与同组第3周比较,中药高、低剂量组大鼠第5、7周肾组织Nephrin mRNA表达显著降低(P<0.05)。【结论】肾炎消白颗粒可通过上调肾小球肾炎蛋白尿模型大鼠肾组织中Nephrin mRNA表达,修复损伤的足细胞,高剂量治疗效果更明显。
Objective To observe the effect of Shenyan Xiaobai Granules on the expression of Nephrin mRNA in renal tissues of glomerulonephritis rats with proteinuria, and to explore its mechanism. Methods One hundred Wistar male rats were randomly divided into blank group,model group,Lotensin group,high-and low-dose Chinese medicine groups, 20 rats in each group. The rat model of glomerulonephritis with proteinuria was established by one-time injection of doxorubicin through tail vein. The rats in Lotensin group were given introgastric administration of 0.90 mg·kg~(-1)·d·~(-1) of Lotensin. The rats in high-and low-dose Chinese medicine groups were given introgastric administration of 3.60,1.80 g·kg~(-1)·d·~(-1) of Shenyan Xiaobai Granules,respectively.The medication lasted 6 continuous days in one week, and the treatment covered 7 weeks. The body mass in various groups was measured once every week. At week 3,5,7,the pathological features of renal tissues in various groups were observed by haematoxylin-eosin(HE)staining method,and Nephrin mRNA expression levels of rat renal tissues were detected by real-time fluorescence quantitative reverse transcription-polymerase chain reaction(RT-q PCR). Results(1)Body mass:Compared with the blank group, the rat body mass had abnormal increase in the model group,particularly after modeling for 3 weeks(P < 0.05). Compared with the model group,the abnormal increase of rat body mass in the Lotensin group at different time points and that in the high-dose Chinese medicine group after modeling for 3 weeks were controlled effectively(P < 0.05),the control effect in the high-dose Chinese medicine group after modeling for 3 weeks being superior to that of the Lotensin group(P <0.05).(2)Ratio of urinary albumin(UA)to urine creatinine(UCr):Compared with the blank group,the ratio of UA to UCr in the model group was increased at different time points(P<0.01). Compared with the model group,the ratio of UA to UCr in Lotensin group at week 1-7,and in high-dose Chinese group at week 3,4,5,6,7 was decreased(P<0.05). Compared with Lotensin group,the ratio of UA to UCr in high-dose Chinese group at week 5,6,7 was decreased(P<0.05).(3)Nephrin mRNA:Compared with the blank group,Nephrin mRNA expression level in the model group was increased at week 3 and was decreased at week 5,7(P<0.05). Compared with the model group,Nephrin mRNA expression level in high-dose Chinese medicine group at different time points was increased(P<0.05). Compared with the same group at week 3,Nephrin mRNA expression level in high-and low-dose Chinese medicine groups at week 5, 7 was decreased(P<0.05). Conclusion Shenyan Xiaobai Granules may repair the damaged podocytes through upregulating the expression of Nephrin mRNA in therenal tissues of glomerulonephritis rats with proteinuria,and the therapeutic effect of high dose of Shenyan Xiaobai Granules is stronger.
引文
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