The effect of puromycin on subepithelial deposits induced by antibodies directed against tubular antigens: a quantitative study.

The mechanism of the GN induced by the intravenous injection of antibodies directed against renal tubular epithelial antigens (RTE) was studied by the use of antibody specifically purified by affinity chromatography. With this reagent RTE antigens could be demonstrated on frozen sections of glomeruli as well as on glomerular cells in culture. The antibodies were radiolabelled, and using paired label techniques glomerular binding could be measured in vivo. In normal rats, approximately 4% of the antibody injected was specifically bound in the glomeruli. Binding was not detectable on proximal tubular brush borders. In rats treated with 100 mg/kg of aminonucleoside of puromycin, a substance known to induce alterations of glomerular epithelial cells, it could be shown that glomerular binding was selectively decreased. These observations indicate that immune deposits in passive Heyman nephitis are formed mainly in situ and that local alteration of the glomerulus can markedly influence deposit formation.     

柴黄益肾方对嘌呤霉素氨基核苷诱导大鼠慢性肾病的治疗作用

目的:研究柴黄益肾方对嘌呤霉素氨基核苷(puromycin aminonucleoside,PAN)诱导大鼠慢性肾病的治疗作用。方法:60只Wistar大鼠随机分为假手术组、模型组、福辛普利钠阳性对照组(0．833 mg·kg-1)、柴黄益肾方小、中、大剂量组(0．55,1．1和2．2 g·kg-1),每组10只,除假手术组外其余各组动物均经颈静脉插管一次性注射PAN 50 mg·kg-1,制作大鼠肾病综合征伴局灶节段性肾小球硬化模型,假手术组给予生理氯化钠溶液。各给药组于手术次日起连续灌胃给药23周。观察药物对大鼠24 h尿蛋白定量及血清总蛋白、白蛋白、血清胆固醇、三酰甘油、血清尿素氮、血清肌酐等生化学指标的影响,对肾组织进行病理学观察并对损害程度进行评分,采用免疫组化的方法,观察柴黄益肾方对α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)和转化生长因子(transforming growth factor-β1,TGF-β1)蛋白在肾脏组织表达的影响。结果:柴黄益肾方能明显抑制大量尿蛋白排泄,提高血清总蛋白和白蛋白含量,改善脂代谢紊乱,降低血清尿素氮,改善肾脏病理损伤,降低α-SMA和TGF-β1蛋白在肾脏组织表达。结论:柴黄益肾方能减轻由PAN诱导的肾组织损伤,改善肾功能障碍和脂蛋白代谢紊乱,降低α-SMA和TGF-β1蛋白在肾脏组织表达。     

Tissue distribution of alpha-tocopherol in nephrotic rats

1. Reactive oxygen species are involved in the pathogenesis of puromycin aminonucleoside (PAN) nephrosis and alpha-tocopherol is one of the major anti-oxidants in the body. 2. In the present study, we measured the levels of alpha-tocopherol by high-performance liquid chromatography in the plasma and in nine tissues of control and nephrotic rats obtained 10 days after either 0.9% saline solution or PAN injection, respectively. 3. In nephrotic rats, alpha-tocopherol levels increased four-fold in plasma; however, the molar ratio of alpha-tocopherol/ cholesterol remained unchanged, suggesting that the increase in alpha-tocopherol content was attributable to an increase in plasma lipid concentration. 4. In nephrotic rats, the alpha-tocopherol/cholesterol ratio increased 1.33-fold in adrenal glands and 1.34-fold in the testis, but remained unchanged in heart, spleen, liver, kidney lung, brain and muscle. 5. These data suggest that, in PAN nephrotic rats, there are alterations in the distribution of alpha-tocopherol and there is no deficiency of alpha-tocopherol in plasma or tissues.     

Experimentally induced parthenogenetic activation of human oocytes

A total of 486 unfertilized, aged human oocytes were exposedto ethanol, calcium ionophore A23187, phorbol ester or puromycinand examined for evidence of activation. Five per cent of controloocytes (3/58) were spontaneously activated. Of the two agentswhich cause the release of intracellular Ca²⁺ ions, Ca²⁺ ionophoreinduced activation of only 16% of unfertilized oocytes, whileethanol did not have any effect. Phorbol ester, a stimulatorof protein kinase C, also resulted in limited activation (14%of oocytes). In contrast, puromycin, an inhibitor of proteinsynthesis, resulted in activation of 91% of the exposed oocytes.It is speculated that puromycin probably inhibits a specificcytostatic factor or factors which are responsible for maintenanceof the metaphase II block. Morphologically activated oocytesusually retained the second polar body and formed subnuclei.The developmental potential of activated oocytes appeared tobe reduced, with only some oocytes capable of a single division.     

Pregnancy following chemical activation of oocytes in a couple with repeated failure of fertilization using ICSI: case report

We report our attempts to achieve a successful pregnancy outcome with calcium ionophore A23187 and puromycin oocyte activation using sperm from a normozoospermic husband of a patient with previous repeated failed fertilization following ICSI. Oocytes from the female partner of a couple with a 4 year history of unexplained primary infertility with repeated failed fertilization following ICSI were used. In the latest ICSI attempt, oocytes were activated by treatment with calcium ionophore (5 min) and puromycin (5 h), then cultured. In this cycle, assisted oocyte activation with calcium ionophore and puromycin after ICSI resulted in a satisfactory fertilization rate (8/12; 66.7%); in prior cycles only one of 71 oocytes (1.4%) was fertilized. The outcome was a Caesarean section delivery of a healthy male infant without congenital abnormalities at 41 weeks, 2 days of gestation. In conclusion, the use of calcium ionophore and puromycin for oocyte activation was found to be a useful method in a case of repeated failed fertilization after ICSI.     

Late changes in the course of mitosis in a synchronized Chinese hamster cell culture after inhibition of RNA and protein synthesis

The effect of inhibition of synthesis of various types of RNA and proteins at different periods of interphase on the course of late mitosis was studied in a synchronized culture of Chinese hamster cells. Analysis of the mitotic index and forms of pathology of division showed that the action of different doses of actinomycin D and puromycin in the first half of interphase produces an identical effect: C-mitoses in the immediate and late waves of cell division. Suppression of synthesis of total cell RNA and proteins in the second half of interphase was accompanied by delay of the cells in metaphase, with scattering of the chromosomes, evidence of a disturbance of the synthesis of the components of the division spindle. It is suggested that proteins (tubulins) and RNA participate as a reserve pool in the organization of the division spindle of late mitosis.Laboratory of Cytology, Institute of Human Morphology, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. P. Avtsyn.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 86, No. 7, pp. 78–81, July, 1978.     

嘌呤霉素肾病鼠尿蛋白量与足突形态改变的关系

目的 :研究嘌呤霉素肾病大鼠尿蛋白排出量与肾小球上皮细胞足突的形态变化之间的相互关系 ,初步探讨蛋白尿的发生机制。　 方法 :30只SD大鼠随机分为实验组和对照组 ,在 2天、5天、10天、15天、2 0天 5个不同时间点应用双缩脲法测定大鼠 2 4h尿蛋白排出量 ,并用透射电镜摄片 ,图像分析及形态计量学的方法 ,研究大鼠在不同时间点的足突形态变化 ,进而分析尿蛋白排出量与足突形态改变之间的相互关系。　 结果 :① 2 4h尿蛋白定量结果示 ,注药 5天后尿蛋白排出量逐渐增多 ,10天时尿蛋白排出量与对照组差异非常显著 (P <0 0 1) ,15天时尿蛋白排出量恢复 ,与 10天比差异亦非常显著 (P <0 0 1) ,且与对照组相比差异仍显著 (P <0 0 5 ) ,至 2 0天尿蛋白排出量与对照组无差别 (P >0 15 )。②注射嘌呤霉素后 ,足突形态呈现逐渐肿胀到融合以后又逐渐呈恢复的变化 ,其中 2～ 2 0天实验组足突宽度 ,体积密度 ,比表面与对照组相比差异有高度显著性 (P <0 0 1) ,表面积密度在2～ 15天与对照组差异显著 (P <0 0 5 ) ,各参数均于 10天达极值。③尿蛋白排出量与足突宽度正相关 (r =0 92 94,P =0 0 2 ) ,与体积密度成正相关 (r =0 95 5 9,P =0 0 11) ,而与比表面呈负相关 (r=- 0 9388,P =0 0 18)。　 结论 :尿     

PI3K/Akt信号通路在肾小球足细胞损伤凋亡中的作用机制

[摘要]目的：初步探讨磷脂酰肌醇3-激酶/蛋白激酶B（PI3K/Akt）信号通路在肾小球足细胞损伤凋亡中的作用机制。方法：采用荧光定量PCR技术、免疫印迹（Western blot）技术及激光共聚焦技术检测肾小球足细胞凋亡相关蛋白的表达和分布情况,流氏细胞仪检测肾小球足细胞凋亡率。结果：嘌呤霉素核苷酸（PAN）损伤足细胞后CD2相关蛋白（CD2AP）分子的表达明显降低,与p85的共定位异常;给予地塞米松（DEX）干预后,CD2AP的表达及分布趋于正常,足细胞磷酸化Akt（p-Akt）及磷酸化糖原合成酶激酶-3β（p-GSK3β）的表达与PAN浓度依赖性的下降,给予DEX干预后,p-Akt及p-GSK3β的表达逐渐恢复;PAN损伤足细胞后,其凋亡率明显升高,DEX干预后凋亡率较PAN组降低（P<0.05）。结论：PI3K/Akt信号通路在肾小球足细胞凋亡中发挥了重要作用,而糖皮质激素可能通过稳定PI3K/Akt信号通路发挥保护肾小球足细胞作用。