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中华妇幼临床医学杂志(电子版) ›› 2009, Vol. 05 ›› Issue (05) : 458 -465. doi: 10.3877/cma.j.issn.1673-5250.2009.05.101

论著

槐杞黄对IgA肾病大鼠蛋白尿及肾组织nephrin及podocin的影响
陆慧瑜, 张巧玲, 蒋小云, 陈丽植, 莫樱, 陈述枚   
  1. 510080 广东广州,中山大学附属第一医院儿科
    中山大学附属第一医院黄埔院区儿科
  • 出版日期:2009-10-01

Effects of Huaiqihuang on Proteinuria and Nephrin, Podocin in Rats With IgA Nephropathy

Hui–yu LU, Qiao–ling ZHANG, Xiao–yun JIANG, Li–zhi CHEN, Ying MO, Shu–mei CHEN   

  1. Department of Pediatrics, First Affiliated Hospital of Sun Yat–sen University, Guangzhou 510080, China
  • Published:2009-10-01
  • Supported by:
    * Project No. 2008B03031070, supported by the Technical Project of Guangdong Province; project No. B2008027, supported by the Medical Science Foundation Project of Guangdong Province
引用本文:

陆慧瑜, 张巧玲, 蒋小云, 陈丽植, 莫樱, 陈述枚. 槐杞黄对IgA肾病大鼠蛋白尿及肾组织nephrin及podocin的影响[J]. 中华妇幼临床医学杂志(电子版), 2009, 05(05): 458-465.

Hui–yu LU, Qiao–ling ZHANG, Xiao–yun JIANG, Li–zhi CHEN, Ying MO, Shu–mei CHEN. Effects of Huaiqihuang on Proteinuria and Nephrin, Podocin in Rats With IgA Nephropathy[J]. Chinese Journal of Obstetrics & Gynecology and Pediatrics(Electronic Edition), 2009, 05(05): 458-465.

目的

观察槐杞黄对IgA肾病(IgA nephropathy, IgAN)大鼠蛋白尿及肾组织nephrin及podocin的影响,探讨槐杞黄对IgA肾病大鼠蛋白尿的治疗作用及其机制。

方法

将24只雄性Sprague–Dawley(SD)大鼠按随机数字表法分为对照组、模型组、槐杞黄组,每组8只。模型组和槐杞黄组建立IgA肾病模型,槐杞黄组给予槐杞黄颗粒治疗。应用定量尿沉渣计数法检测尿红细胞个数,考马斯亮蓝法检测24 h尿蛋白定量,全自动生化分析仪检测血生化指标。应用直接免疫荧光法检测肾小球IgA沉积强度,间接免疫荧光法检测肾小球nephrin及podocin蛋白的表达及分布,采用实时荧光定量PCR技术检测肾皮质nephrin mRNA和podocin mRNA的表达。

结果

①槐杞黄组大鼠的尿红细胞计数、尿蛋白,较模型组显著减少,差异有显著意义(P<0.05);②槐杞黄组大鼠肾组织病变,较模型组减轻;③槐杞黄组大鼠肾小球nephrin表达量,较模型组显著升高,差异有显著意义(P<0.01),podocin表达量较模型组稍有下降,但差异无显著意义(P>0.05);槐杞黄组大鼠肾皮质nephrin mRNA及podocin mRNA的表达,较模型组显著升高,差异有显著意义(P<0.01);④槐杞黄组大鼠肾小球nephrin及podocin的不连续斑片状、团块状异常分布,较模型组改善。

结论

IgA肾病大鼠肾组织nephrin及podocin出现表达变化及分布异常,槐杞黄能降低IgAN大鼠尿蛋白,其作用可能与调节nephrin及podocin的表达及分布有关。

Objective

To observe effects of huaiqihuang on proteinuria in rats with IgA nephropathy (IgAN) and dynamic changes of expression and distribution of nephrin, podocin, and to further explore the mechanism of proteinuria of IgA nephropathy and try to find the approach to the therapy for IgA nephropathy.

Methods

A total of 24 Sprague–Dawley (SD) rats with 6 weeks (n=24) were randomLy divided into 3 groups: Control group (n=8), model group (n=8) and huaiqihuang group (n=8). IgA nephropathy model was induced by the improved method of bovine serum albumin (BSA)+ carbon tetrachloride (CCl4)+ lipopolysaccharide (LPS). At the end of the experiment, hematuria, proteinuria and renal pathology should be noticed. The IgA intensity was examined by direct immunofluorescence. The protein expression and distribution of nephrin and podocin were examined by indirect immunofluorescence. The mRNA expression of nephrin and podocin were examined by real–time PCR.

Results

① The count of urinary red cells and urinary protein in huaiqihuang group were significant less than those of model group (P<0.05). ②Mesangial proliferation, foot processes fusion and IgA deposition in glomeruli of huaiqihuang group were less serious than those of model group. ③The protein expressions of nephrin in huaiqihuang group were significant higher than those in model group (P<0.01). Podocin protein expression in huaiqihuang group had no significant difference from those of model group (P>0.05), but were still higher than those of control group. In huaiqihuang group, and mRNA of nephrin and podocin expressed markedly higher than those of model group (P<0.01). ④Abnormal discontinuous distribution of nephrin and podocin in huaiqihuang group was slighter than that in model group.

Conclusion

Huaiqihuang is good for decreasing urinary protein by regulating the expression and distribution of nephrin and podocin.

图1 Nephrin、podocin及内参GAPDH的实时荧光定量PCR熔解曲线图
图2 各组大鼠肾小球形态学改变(HE染色、PAS染色、Masson三色染色,光镜×400)
图3 各组大鼠肾组织透射电镜图
图4 各组大鼠肾小球IgA沉积强度(免疫荧光,×200)
图5 各组大鼠肾小球nephrin免疫荧光图(免疫荧光,×200)
图6 各组大鼠肾小球podocin免疫荧光图(免疫荧光,×200)
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