Freshly isolated renal cells as a experimental model system for studying the chemically induced renal injury.

新鲜分离的肾细胞作为研究化学诱导的肾损伤的实验模型系统。

• 批准号: 59480124
• 负责人: TAKAHASHI Atsushi
• 金额: $ 4.35万
• 依托单位: National Institute of Hygienic Sciences.
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1984
• 资助国家: 日本
• 起止时间: 1984 至 1986
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-59480124/
• 关键词: Renal cells; Freshly isolated renal cells; 腎毒性; 実験モデル

For the purpous to establish freshly isolated renal cells as a in vitro test method of toxicity evaluation, we improved the method of Jones'(1979) and obtained the cells with high viability ( >95%) and yield (Ca <10^8> cells/rat). This preparation had a high activity of the enzymes which existed predominantly in proximal renal tubular epithelial cells. Most of the cells bound with monoclonal antibody which was specific to the epithelial cells. These results indicated that most of the cells were originated from this region of the kidneys. Parathyroid hormon increased cAMP content of the renal cells.Carriar of the organic anion was shown to exist in the cell membrane using 8-anilino 1-naphthalene sulfornate. Aminopyrine metabolizing activities of the renal cells were about 0.4% of those of hepatocytes. Toxic effects of chemicals were : 1 ) Direct acting agents, such as <HgCl_2> , <CdCl_2> , and disulfiram, caused the loss of viability of the cells. Although the effects of disulfiram were inhibited by GSH, those of <CdCl_2> were not inhibited completely by cysteine and GSH. 2 ) Effects of gentamycin were not observed, but adriamycin decreased the GSH content of the cells. 3 ) Cyclophosphamide and paracetamol, which were known to be activated metabolically by cyt. P-450 linked drug metabolizing system, did not have toxic effects. 4 ) Toxicity of allyl alcohol were marked in female rats, in which activity of alcohol dehydrogenase were higher than in male rats.These results indicated that the renal cell preparation could be used to study the mechanism of acute toxicity of chemicals.

为了建立一种新的肾细胞毒性评价的体外试验方法，我们改进了Jones（1979）的方法，获得了高活性（>95%）和高产量（Ca <10 ^8> cells/rat）的细胞。该制剂具有主要存在于近端肾小管上皮细胞中的酶的高活性。大多数细胞与上皮细胞特异性的单克隆抗体结合。这些结果表明，大多数细胞来源于肾脏的这个区域。甲状旁腺素使肾细胞cAMP含量增加，用8-苯胺基1-萘磺酸盐证明细胞膜上有有机阴离子的载体。肾细胞的氨基比林代谢活性约为肝细胞的0.4%。化学品的毒性作用有：1）直接作用剂，如<HgCl_2>，<CdCl_2>和双硫仑，导致细胞活力丧失。双硫仑的作用可被GSH抑制，但<CdCl_2>半胱氨酸和GSH不能完全抑制双硫仑的作用。（2）庆大霉素对细胞的GSH含量无明显影响，而阿霉素可使细胞GSH含量降低。3）环磷酰胺和扑热息痛，已知它们被cyt代谢活化。P-450连接药物代谢系统，没有毒性作用。（4）烯丙醇对雌性大鼠有明显的毒性作用，其乙醇脱氢酶活性明显高于雄性大鼠，表明该肾细胞制备方法可用于研究化学品急性毒性的机制。

项目成果

Ohno Y.: "Evaluation of the toxicity of chemicals using freshly isolated liver and renal cells." Toxicology Forum.10. (1987)

Ohno Y.：“使用新鲜分离的肝和肾细胞评估化学品的毒性。”

Ohno Y., Takahashi A., Kawanishi T., Takanaka A., Omori Y.: "Origin and characteristics of freshly isolated renal cells obtained by the perfusion of collagenase in vitro." J. Pharmacobio-Dynamics. 9. s-66 (1986)

Ohno Y.、Takahashi A.、Kawanishi T.、Takanaka A.、Omori Y.：“通过体外灌注胶原酶获得的新鲜分离肾细胞的起源和特征。”

大野泰雄,川西徹,高橋惇,高仲正,大森義仁: J.Pharmacobio-Dynamics. 9. -66 (1986)

Yasuo Ohno、Toru Kawanishi、Atsushi Takahashi、Masaru Takanaka、Yoshihito Omori：J.Pharmacobio-Dynamics 9. -66 (1986)。

大野泰雄: トキシコロジーフォーラム. 10. (1987)

大野康夫：毒理学论坛 10。（1987）