Protective effect of tetramethylpyrazine on kidney in rats with membranous nephropathy

doi:10.3969/j.issn.2095-4344.1100

Abstract

Abstract:

BACKGROUND: Membranous nephropathy, a common type of nephrotic syndrome in adults, is characterized by podocyte decrease caused by apoptosis. Podocyte apoptosis is regulated by Bcl-2 and Bax. Tetramethylpyrazine exerts good effectiveness in the treatment of renal diseases.
OBJECTIVE: To observe the protective effect of tetramethylpyrazine on cationized bovine serum albumin-induced membranous nephropathy in rats.
METHODS: Seventy-two rats provided by Laboratory Animal Center of Southwest University were randomly divided into normal, model and tetramethylpyrazine groups (n=24 per group). Rabbits in the latter two groups underwent membranous nephropathy modeling by modified Border method. The tetramethylpyrazine group received intraperitoneal injection of 100 mg/kg tetramethylpyrazine for four consecutive weeks.
RESULTS AND CONCLUSION: Compared with the normal group, the model group showed strong IgG-like deposition in the glomerular mesangial area and capillary wall; glomerular volume increased, as shown by light microscopy. Masson staining showed chlorocruorin deposition. Gomori methenamine silver staining showed diffuse thickening of basement membrane and formation of "nail spikes". Electron microscope showed thickening of the glomerular basement membrane, fusion of foot process, and deposition of electron dense deposits on the glomerular epithelium. Compared with the model group, there was no significant difference in the fluorescence intensity and distribution of IgG in the tetramethylpyrazine group, but the degree of glomerular basement membrane thickening and the degree of foot process fusion were slightly improved. Compared with the normal group, the serum creatinine and 24-hour urinary protein levels were increased, creatinine clearance was decreased, expression level of Bcl-2 and the number of WT-1 positive cells were increased, and Bax expression level and the number of apoptotic cells were decreased in the other two groups. To conclude, tetramethylpyrazine can improve the renal function of rats with membranous nephropathy induced by cationized bovine serum albumin and increase serum albumin. It protects the kidney by reducing the cell apoptosis through regulating Bcl-2 and Bax pathways and inhibiting the reduction of podocytes.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: Nephrosis, Apoptosis, Tissue Engineering

CLC Number:

R453
R364

Cheng Runxia1, Liang Jing2, Liu Gang3, Yang Linlin1, Zhang Yao4, Cao Ling1. Protective effect of tetramethylpyrazine on kidney in rats with membranous nephropathy[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(11): 1730-1737.

Figures/Tables 5

2.3 川芎嗪可减轻膜性肾病大鼠的部分肾脏病理改变免疫荧光第4周末时，正常组肾组织无异常。模型组可见肾小球系膜区及毛细血管壁IgG强阳性颗粒样沉积；光镜下可见肾小球体积增大，马松染色见部分上皮下大量嗜复红蛋白沉积，六胺银染色可见基底膜弥漫性增厚、“钉突”形成，部分肾小管上皮细胞肿胀变性；电镜下可见肾小球基底膜明显增厚，足细胞的足突弥漫性融合，肾小球上皮下大量电子致密物沉积。与模型组相比，川芎嗪组肾小球IgG荧光强度及分布无明显差异，但光镜或电镜下肾小球基底膜增厚的程度及足突融合程度都有轻微改善，见图2。 2.4 川芎嗪显著减轻膜性肾病大鼠肾实质细胞的凋亡凋亡细胞主要分布在肾小管上皮细胞，其次分布在肾小球脏层、肾小球系膜区。与正常组大鼠肾组织相比，模型组和川芎嗪组肾组织中凋亡细胞明显增多(P < 0.05)，从第1周末开始在肾小球及肾小管内的凋亡细胞均增加(P < 0.05)；模型组和川芎嗪组的细胞凋亡数随时间的延长逐渐增多，正常组无此变化。与模型组相比，川芎嗪组从第1周末开始在肾小球及肾小管内的凋亡细胞较少(P < 0.05)，见图3。 2.5 川芎嗪可影响膜性肾病大鼠肾组织中Bcl-2，Bax及WT-1的表达水平 2.5.1 肾小球和肾小管中Bcl-2表达水平、Bax表达水平与Bax/Bcl-2比值正常组大鼠肾组织中Bcl-2蛋白表达于肾小球脏层上皮细胞、节段系膜区和肾小管上皮细胞胞浆中，且随时间推移无明显变化，但模型组及川芎嗪组各时间点的Bcl-2表达水平比正常组明显减少(P < 0.05)，表达部位无明显差异，且随时间的延长，2组中Bcl-2表达水平均呈逐渐下降趋势；与模型组相比，川芎嗪组Bcl-2在肾小球及肾小管内的表达水平略有上升(第3，4周末时P < 0.05)，见图4。正常组大鼠肾组织中Bax在肾小球系膜区及肾小管上皮细胞胞浆仅有极少量表达，而模型组及川芎嗪组肾组织中Bax在肾小球脏层上皮细胞、肾小球系膜区、肾小管上皮细胞浆内均有表达，且表达水平明显高于正常组(P < 0.05)，且随时间的延长，Bax表达水平呈上升趋势；与模型组相比，川芎嗪组Bax表达水平在肾小球及肾小管内较低，尤其以第3，4周末差异有显著性意义(P < 0.05)，见图5。此外实验还发现，模型组和川芎嗪组较正常组肾组织中Bax/Bcl-2比值升高(P < 0.05)；且随时间的延长，Bax/Bcl-2比值水平逐渐升高；与模型组相比，川芎嗪组Bax/Bcl-2比值较低(P < 0.05)，见图6。 2.5.2 肾小球中WT-1阳性细胞比例 WT-1分布于肾小球足细胞核内。与正常组相比，模型组和川芎嗪组肾小球内的WT-1阳性细胞比例减少(P < 0.05)；且随时间的延长，2组肾小球内的WT-1阳性细胞比例都进一步下降；与模型组相比，川芎嗪组WT-1阳性细胞比例较多，其中第3，4周末差异有显著性意义(P < 0.05)，见图7。 2.6 模型组大鼠肾组织内细胞凋亡数量与Bcl-2表达水平、Bax表达水平、Bax/Bcl-2、WT-1阳性细胞比例的相关性模型组大鼠肾小球的细胞凋亡数量与Bax表达水平、Bax/Bcl-2比值呈正相关(P < 0.05)，与Bcl-2表达水平、WT-1阳性细胞比例呈负相关(P < 0.05)，而模型组大鼠肾小管中细胞凋亡数与Bax表达水平、Bax/Bcl-2比值呈正相关(P < 0.05)，与Bcl-2表达水平呈负相关(P < 0.05)，见图8。"

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