中国血液净化

›› 2008, Vol. 7 ›› Issue (4): 198-202.

• 基础研究 • 上一篇    下一篇

去甲斑蝥素对蛋白超负荷肾病大鼠CTGF及NF-κBp65表达的影响

叶琨 刘伏友 彭佑铭   

  1. 中南大学湘雅二医院肾内科
  • 收稿日期:2008-01-22 修回日期:1900-01-01 出版日期:2008-04-12 发布日期:2008-04-12
  • 通讯作者: 叶琨

Effect of norcantharidin on expression of connective tissue growth factor and nuclear factor-闎p65 in kidney of protein-overloaded nephropathy rats

YE Kun, LIU Fu-you, LI Ying   

  1. PENG You-ming, DUAN Shao-bin, XU Xiang-qing, ZHOU Le-tian, LI Zhi-lan. Department of Nephrology, The Second Xiangya Hospital, Central South University, Changsha 410011, China
  • Received:2008-01-22 Revised:1900-01-01 Online:2008-04-12 Published:2008-04-12

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摘要： 目的 观察去甲斑蝥素对蛋白超负荷肾病大鼠结缔组织生长因子（CTGF）及核因子-κB（NF-κB）p65 mRNA及蛋白表达的影响，初步探讨其延缓肾间质纤维化的机制。 方法 所有SD大鼠均行单侧肾切除术。实验组大鼠予以腹腔注射牛血清白蛋白（BSA），建立蛋白超负荷肾病模型后随机分为模型组（BSA组，腹腔注射BSA 5g·kg-1·d-1）和去甲斑蝥素组（NCTD组，腹腔注射BSA 5g·kg-1·d-1和NCTD 0.1mg·kg-1·d-1）；对照组（Saline组，腹腔注射等量生理盐水）；每组8只大鼠。第9周末处死大鼠，留取肾脏标本。行光镜和电镜观察肾组织病理改变；RT－PCR法检测CTGF和NF-κBp65 mRNA表达；Western blot及免疫组化法分别检测CTGF和NF-κBp65 蛋白表达。 结果 ⑴肾组织病理改变：BSA组可见肾小球球性硬化，肾小管灶性萎缩，肾间质大量炎症细胞浸润，伴灶状纤维化，病理半定量积分显示小管间质病理损伤（TIL）积分高于Saline组（P＜0.01）；电镜示肾小球系膜区有电子致密物沉积，足突融合。NCTD干预后，肾组织上述病理改变有所改善，且该组TIL积分较BSA组降低（P＜0.05）。⑵RT－PCR结果：NCTD组CTGF及NF-κBp65 mRNA的表达较BSA组均减弱（P＜0.01）。⑶Western blot检测结果提示NCTD组CTGF蛋白表达较BSA组下调（P＜0.05）；免疫组化显示，与BSA组比较，NCTD组NF-κBp65表达分布的阳性面积和阳性细胞数均明显降低（P＜0.05）。 结论 去甲斑蝥素可通过下调蛋白超负荷肾病大鼠CTGF和NF-κBp65的表达水平，延缓肾间质纤维化。

关键词: 斑蝥素, 肾间质纤维化, 结缔组织生长因子, 核因子-κB

Abstract: Objective To detect expression of connective tissue growth factor (CTGF) and nuclear factor-κB (NF-κB)p65 in renal tissues，and investigate the mechanism of Norcantharidin (NCTD) on renal interstitial fibrosis prevention. Methods uninephrectomized SD rats were received intraperitoneally injections of bovine serum albumin (BSA). The nephropathy rats were randomly divided into model group (BSA group, intraperitoneally injections of BSA 5g·kg-1·d-1) and NCTD group (intraperitoneally injections of BSA 5g·kg-1·d-1and NCTD 0.1mg·kg-1·d-1). An additional set of uninephrectomized rats were only given saline as control group. At week 9, the rats were sacrificed and the remnant kidney was collected. Renal pathological change was observed by light microscopy and electron microscopy. Expression of CTGF mRNA and NF-κBp65 mRNA in renal tissue was semi-quantitative analyzed by RT-PCR. CTGF protein expression was detected by western blot. NF-κBp65 protein expression was detected by immunohistochemistry. Result ⑴By light microscope, in BSA group it was showed that glomerular sclerosis existed and lots of inflammatory cell infiltrated in renal interstitium with segmental interstitial fibrosis. By electron microscope, obvious electron dense deposits existed and foot process fusion was seen extensively in BSA group. After intervention of NCTD, tubulointerstitial lesion（TIL）score significantly decreaded (P＜0.05). ⑵Compared with BSA group, expression of CTGF mRNA and NF-κBp65 mRNA in NCTD group were down-regulated (P＜0.01). ⑶Compared with BSA group, protein expressions of CTGF in NCTD was down-regulated (P＜0.05) by western blot. ⑷Compared with BSA group, deposited area and positive cell population of NF-κBp65 decreased (P＜0.05). Conclusion NCTD could effectively prevent renal tubulointerstitial fibrosis by significantly down-regulating expressions of CTGF and NF-κBp65.

Key words: Renal tubulointerstitial fibrosis, Connective tissue growth factor, Nuclear factor-κB p65

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