分泌型卷曲相关蛋白4在常染色体显性遗传性多囊肾病中的表达及诱发凋亡的作用被引量：1

Expression of secreted frizzled related protein 4 and effect of induced-apoptosis in autosomal dominant polycystic kidney disease

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摘要目的观察分泌型卷曲相关蛋白4（sFRP4）在常染色体显性遗传性多囊肾病（ADPKD）中的表达情况及其在ADPKD中诱发凋亡的作用。方法（1）血清：收集12例健康人和20例ADPKD患者的血清标本，ELISA法检测血清中sFRP4的水平。（2）组织：收集肾癌根治性切除术后癌旁正常肾组织以及ADPKD患者肾组织，免疫组化法观察sFRP4在肾组织中的表达情况；实时定量PCR法检测肾组织中sFRP4mRNA及Caspase-3 mRNA水平；TUNEL法检测肾组织细胞凋亡。（3）细胞：HEK-293T细胞被分为3组：空白对照组、对照质粒PcDNA6转染组和Flag-sFRP4．PcDNA6质粒转染组，Western印迹法检测凋亡相关蛋白Caspase．3表达水平，流式细胞术检测各组细胞凋亡率。结果（1）ELISA检测结果显示，ADPKD患者血清sFRP4水平显著高于健康人（P〈0．05）。（2）免疫组化结果显示，sFRP4在ADPKD患者的肾小管上皮细胞胞质中表达明显增加。（3）实时定量PCR结果显示，ADPKD患者肾组织sFRP4mRNA和Caspase-3 mRNA表达水平显著高于正常肾组织（均P〈0．05）。（4）TUNEL检测结果显示，ADPKD患者肾小管上皮细胞凋亡较正常肾组织明显增加。（5）与对照组相比，过表达sFRP4的HEK-293T细胞中Caspase-3蛋白水平显著上调（P〈0．05），细胞凋亡率显著升高（P〈0．05）。结论sFRP4在ADPKD患者血液及肾组织中均明显升高，ADPKD中存在肾小管上皮细胞的异常凋亡，过表达sFRP4可以诱导HEK-293T细胞凋亡，推测sFRP4升高与ADPKD肾组织的异常凋亡增加有关。 Objective To evaluate the expression of secreted frizzled related protein 4 （sFRP4） in autosomal dominant polyeystic kidney disease（ADPKD） and the effect of sFRP4 induced apoptosis in ADPKD. Methods （1）Serological method： serum samples of 12 healthy people and 20 ADPKD patients were collected and the levels of sFRPd in serum were detected by ELSIA assay. （2） Tissue experiments： normal renal tissue was collected from radical nephrectomy for renal carcinoma; polycystic renal tissues were taken from ADPKD patients. The expression of sFRP4 in renal tissues was observed by immunohistochemistry; Real-time PCR was used to explore the mRNA level of sFRP4 and caspase-3; TUNEL method was applied to observe the apoptosis cells existing in ADPKD. （3）studies in vitro： HEK- 293T cells were transfected with PcDNA6 and Flag. sFRP4.PcDNA6 respectively, after which Western blotting was performed to detect the expression of caspase-3 protein and flow cytometry was performed to estimate cell apoptosis rate. Results （1）ELISA results showed serum concentrations of sFRP4 in ADPKD were markedly higher than normal control （P 〈 0.05）. （2）Compared with normal renal tissues, the sFRP4 expression was dramatically increased in ADPKD and mainly distributed in the cytoplasm of renal tubular epithelial cells. （3）Real-time PCR showed the expression of sFRP4 and Caspase-3 mRNA in ADPKD were up-regulated comparing with those in normal control （P 〈 0.05）. （4）TUNEL assays revealed that elevated apoptosis appeared in tubular epithelial cells of ADPKD. （5）The level of caspase-3 protein and apoptosis rate were significantly increased after over-expressed sFRP4 in HEK- 293T cells （all P 〈 0.05）. Conclusions The expression of sFRP4 is strikingly up-regulated in ADPKD. In addition, abnormal apoptosis of tubular epithelial ceils exists in ADPKD and over-expressed sFRP4 can induce apoptosis of HEK-293T cells. This phenomenon may be attributed to the elevated sFRP4.

作者杜宇裴广畅覃海澳吕永曼李月强代维李俊华何凡刘颜颜徐钢高红宇

机构地区华中科技大学同济医学院附属同济医院肾病内科华中科技大学同济医学院附属同济医院综合医疗科

出处《中华肾脏病杂志》 CAS CSCD 北大核心 2016年第6期430-435,共6页 Chinese Journal of Nephrology

基金国家自然科学基金（81100481）

关键词多囊肾常染色体显性细胞凋亡半胱氨酸天冬氨酸蛋白酶3 分泌型卷曲相关蛋白4 Polycystic kidney, autosomal dominant Apoptosis Caspase 3 Secreted frizzled related protein 4

分类号 R692 [医药卫生—泌尿科学]

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