Cytosolic free calcium and intracellular pH in ischemic and drug-induced acute renal failure.

缺血性和药物引起的急性肾衰竭中的胞浆游离钙和细胞内 pH 值。

• 批准号: 01570366
• 负责人: ASANO Yasushi
• 金额: $ 1.41万
• 依托单位: Jichi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-01570366/
• 关键词: Ishemic acute; Drug-induced nephrotoxicity; Cytorolic-free calcium; Intracellular pH; Superoxide

1. Cytosolic free calcium ([Ca^<2+>]i) and intracellular pH(pHi) during cell ischemiaThe effect of ischemia on [Ca^<2+>]i and pHi was determined in cultured rat inner medullary collecting duct (IMCD) cells exposed to NaCN (5mM) and 2-deoxy-D-glucose (5mM). After the addition of cyanide plus 2-deoxy-D-glucose, [Ca^<2+>]i rose and pHi decreased (DELTApH=-0.35). Verapamil (10^<-4>M) or EGTA (2mM) significantly blunted the rise in [Ca^<2+>]i, and decreased pHi moreover (DELTApH=-0.65). Lowering extracellular pH from 7.8 to 6.4 inhibited the rise in [Ca^<2+>]i induced by metabobic inhibitors. At pH 6.5, or with verapamil IMCD cells were protected against ischemic injury as compared with cells incubated at pH 7.4.2. Effect of cyclosporin on [Ca^<2+>]i.Cyclosporin (10^<-8>M) had no effect on [Ca^<2+>]i in LLC-PK, cells.3. Oxidative stress in cultured rat IMCD cells : role of Ca^<2+>.Tert-Butyl Hydroperoxide (TBHP) (5mM) increased [Ca^<2+>]i and reduced IMCD cells viability to 26i, <plus-minus>8.9% at 60min. The increase in [Ca^<2+>]i induced by TBHP was significantly inhibited by EGTA (2mM) or verapamil (10^<-4>M). Both EGTA and verapamil provided significant protection (cell viability. TBHP+EGTA : 61.7<plus-minus>13.1%, TBHP + verapamil : 64.7<plus-minus>5.5%, TBHP : 26.7 <plus-minus>8.9%. P<0.001>. Lowering extracellular pH from 7.4 to 6.6 blunted the rise in [Ca^<2+>] induced by TBHP and protected cell injury (Cell viability. at pH 6.6 : 66.2 <plus-minus> 15.7%)

1.细胞缺血期间的胞质游离钙([Ca^2+]i)和细胞内pH(pHi)在暴露于NaCN(5mM)和2-脱氧-D-葡萄糖(5mM)的培养的大鼠内髓集合管(IMCD)细胞中测定缺血对[Ca^2+]i和pHi的影响。添加氰化物加2-脱氧-D-葡萄糖后，[Ca 2+ ] i 上升且pHi下降(DELTApH＝-0.35)。维拉帕米(10^-4M)或EGTA(2mM)显着抑制了[Ca^2+]i的上升，并且还降低了pHi(DELTApH＝-0.65)。将细胞外pH从7.8降低至6.4抑制代谢抑制剂诱导的[Ca 2+ ] i 的升高。与在 pH 7.4.2 下孵育的细胞相比，在 pH 6.5 下或使用维拉帕米 IMCD 细胞可免受缺血性损伤。环孢素对[Ca ^ 2+ ] i 的影响。环孢素(10 ^ -8 M)对LLC-PK细胞中的[Ca ^ 2+ ] i 没有影响。3．培养的大鼠IMCD细胞中的氧化应激：Ca^2+.叔丁基过氧化氢(TBHP)(5mM)的作用增加[Ca^2+]i并将IMCD细胞活力降低至26i，60分钟时<正负>8.9％。 TBHP诱导的[Ca 2+ ] i 增加被EGTA(2mM)或维拉帕米(10 ^ -4 M)显着抑制。 EGTA 和维拉帕米均提供显着的保护（细胞活力。TBHP+EGTA：61.7<正负>13.1%，TBHP+维拉帕米：64.7<正负>5.5%，TBHP：26.7<正负>8.9%。P<0.001>。将细胞外 pH 从 7.4 降低至 6.6 减弱了细胞活性。上升 TBHP 诱导的 [Ca^2+] 并保护细胞损伤（细胞活力。在 pH 6.6 时：66.2 <正负> 15.7%）

项目成果

Tomoko Ohara: "Regulatoty mechanism on intracecllular pH in inner medullary collecting duct cells in culture:role of Ca^<2+> cAMP and protein Kinase C."

Tomoko Ohara：“培养的内髓集合管细胞内 pH 值的调节机制：Ca^2 cAMP 和蛋白激酶 C 的作用。”

大原智子,武田和司,浅野泰: "ラット培養乳頭部集合管(PCD)細胞における細胞内pH〔pHi〕の調整機序" 第32回日本腎臓学会総会 予稿集. 295 (1989)

Tomoko Ohara、Kazushi Takeda、Yasushi Asano：“培养的大鼠乳头集合管 (PCD) 细胞中细胞内 pH [pHi] 的调节机制”日本肾病学会第 32 届年会记录 295 (1989)。

Takeda K.,Ohara T.,Tabei K.and Asano Y.: "Dual regulatory mechanisms of proton transport in rat papillary collecting duct cells in culture." Jpn.J.Physiol.39. 397-410 (1989)

Takeda K.、Ohara T.、Tabei K. 和 Asano Y.：“培养的大鼠乳头集合管细胞中质子运输的双重调节机制。”

Takeda, K., Terao N., Ohara T., and Asano Y.: "Oxidative stress in cultured rat inner medullany duct cells."

Takeda, K.、Terao N.、Ohara T. 和 Asano Y.：“培养的大鼠内髓管细胞中的氧化应激。”

Ohara T., Takeda K., Tabei K. and Asano Y.: "Effect of ATP depletion o intracellular pH and Cytosolic free calcium in rat inner medullary collecting duct cells."

Ohara T.、Takeda K.、Tabei K. 和 Asano Y.：“ATP 消耗对大鼠内髓集合管细胞内 pH 值和胞质游离钙的影响。”