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血管加压素刺激A10血管平滑肌细胞中[3H] - 肌醇磷酸和[3H] - 磷脂丁醇的积累。

Vasopressin-stimulated [3H]-inositol phosphate and [3H]-phosphatidylbutanol accumulation in A10 vascular smooth muscle cells.

作者信息

Plevin R, Stewart A, Paul A, Wakelam M J

机构信息

Department of Biochemistry, University of Glasgow.

出版信息

Br J Pharmacol. 1992 Sep;107(1):109-15. doi: 10.1111/j.1476-5381.1992.tb14471.x.

DOI:10.1111/j.1476-5381.1992.tb14471.x
PMID:1330154
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1907616/
Abstract
  1. The characteristics of vasopressin-stimulated phosphatidylinositol 4,5 bisphosphate (PtdIns(4,5)P2) and phosphatidylcholine (PtdCh) hydrolysis were examined in A10 vascular smooth muscle cells (VSMC), by assessing the formation of [3H]-inositol phosphates ([3H]-IP) and the accumulation of the phospholipase D (PLD) specific product, [3H]-phosphatidylbutanol ([3H]-PtdBuOH). 2. Vasopressin ([Arg8]-VP) and a number of related analogues stimulated the accumulation of [3H]-IP and [3H]-PtdBuOH with similar EC50 values, generating the same rank order of potency for each response (Arg8-VP = vasotocin = Lys8-VP much greater than oxytocin). 3. Inhibition of vasopressin-stimulated [3H]-IP and [3H]-PtdBuOH accumulation by the V1a receptor antagonists, Des-Gly9[beta-mercapto-beta,beta,-cyclopentamethylene propionyl, O-Et-Tyr2,Val4,Arg8]-vasopressin generated similar IC50 values suggesting that both these responses are mediated through the activation of a single V1a receptor subtype. 4. The onset of vasopressin-stimulated inositol-1,4,5-trisphosphate (Ins(1,4,5)P3) mass formation preceded [3H]-PtdBuOH accumulation indicating that PtdCh hydrolysis was activated subsequent to PtdIns(4,5)P2 breakdown. 5. The protein kinase C (PKC) activator, tetradecanoylphorbol acetate (TPA) also stimulated [3H]-PtdBuOH accumulation. Preincubation with the PKC inhibitor Ro-31-8220 abolished both TPA- and vasopressin-stimulated [3H]-PtdBuOH, suggesting that the intermediate activation of protein kinase C is involved in the regulation of PLD by vasopressin. 6. Pretreatment of the A10 VSMC with Ro-31-8220 (100 microM) also potentiated vasopressin-stimulated Ins(1,4,5)P3 mass formation.Therefore stimulation of PKC may have opposing roles in the regulation of agonist activation of PLC and PLD.7. Preincubation of the cells with EGTA, verapamil, or the receptor-operated calcium channel antagonist, SK&F 96365, reduced vasopressin-stimulated [3H]-PtdBuOH accumulation by approximately 30%, suggesting that influx of calcium has a significant role to play in the regulation of vasopressinstimulated PLD activity.
摘要
  1. 通过评估[3H]-肌醇磷酸酯([3H]-IP)的形成以及磷脂酶D(PLD)特异性产物[3H]-磷脂丁醇([3H]-PtdBuOH)的积累，研究了血管加压素刺激的磷脂酰肌醇4,5-二磷酸(PtdIns(4,5)P2)和磷脂酰胆碱(PtdCh)水解在A10血管平滑肌细胞(VSMC)中的特征。2. 血管加压素([Arg8]-VP)及一些相关类似物刺激[3H]-IP和[3H]-PtdBuOH的积累，其半数有效浓度(EC50)值相似，对每种反应产生相同的效价顺序(Arg8-VP = 血管紧张素 = Lys8-VP远大于催产素)。3. V1a受体拮抗剂去甘氨酸9[β-巯基-β,β-环亚戊基丙酰基，O-乙基-Tyr2,Val4,Arg8]-血管加压素对血管加压素刺激的[3H]-IP和[3H]-PtdBuOH积累的抑制产生相似的半数抑制浓度(IC50)值，表明这两种反应均通过单一V1a受体亚型的激活介导。4. 血管加压素刺激的肌醇-1,4,5-三磷酸(Ins(1,4,5)P3)质量形成先于[3H]-PtdBuOH积累，表明PtdCh水解在PtdIns(4,5)P2分解后被激活。5. 蛋白激酶C(PKC)激活剂十四酰佛波醇乙酸酯(TPA)也刺激[3H]-PtdBuOH积累。用PKC抑制剂Ro-31-8220预孵育可消除TPA和血管加压素刺激的[3H]-PtdBuOH，提示蛋白激酶C的中间激活参与血管加压素对PLD的调节。6. 用Ro-31-8220(100μM)预处理A10 VSMC也增强了血管加压素刺激的Ins(1,4,5)P3质量形成。因此，PKC的刺激在激动剂对PLC和PLD的激活调节中可能具有相反的作用。7. 用乙二醇双四乙酸(EGTA)、维拉帕米或受体操纵性钙通道拮抗剂SK&F 96365预孵育细胞，使血管加压素刺激的[3H]-PtdBuOH积累减少约30%，提示钙内流在血管加压素刺激的PLD活性调节中起重要作用。

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Vasopressin and angiotensin II receptors in rat aortic smooth muscle cells in culture.培养的大鼠主动脉平滑肌细胞中的血管加压素和血管紧张素II受体
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