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目的研究胰岛素样生长因子-1(Insulin like growth factor-1,IGF-1)对血管紧张素Ⅱ(AngⅡ)诱导的人脐静脉内皮细胞(HUVEC)细胞活力及组织因子(TF)的影响和其作用机制。方法分别用不同浓度 IGF-1预处理 HUVEC 细胞株 HUVEC-12细胞30 min 后加入AngⅡ10~(-6)mol/L 孵育24 h,观察细胞活力及 AngⅡ1型受体(AT1-R)mRNA 变化的量效关系。选择一个作用最明显的 IGF-1浓度预处理 HUVEC-12细胞30 min 后加入AngⅡ10~(-6)mol/L 孵育12～48 h,观察细胞活力变化的时效关系。同时测定孵育24 h 的 HUVEC 裂解液 TF 含量、一氧化氮合酶(NOS)活性及一氧化氮(NO)浓度,并于 NOS 抑制剂(L-NAME)预处理细胞30 min 后,再加入 IGF-1和 Ang Ⅱ,观察孵育24 h 细胞活力、TF、AT1-R mRNA、NOS 及 NO 水平变化。结果①Ang Ⅱ能降低 HUVEC-12细胞活力,上调 AT1-RmRNA 表达及增加 TF 含量,抑制 NOS 活性、减少 NO 生成;②IGF-1能显著抑制 Ang Ⅱ诱导的细胞活力降低和 AT1-R mRNA 表达增加,以0.5μg/ml 浓度作用最明显,此浓度处理细胞12～48 h 内细胞活力恢复具有时间依赖性;③IGF-1能抑制 Ang Ⅱ诱导的 TF 增加及改善 Ang Ⅱ诱导的 NOS 活性降低,增加NO 生成;④L-NAME 可明显拮抗 IGF-1对血管内皮的保护作用。结论 IGF-1可明显提高 Ang Ⅱ诱导的内皮细胞活力及抗血栓能力,其作用可能是通过激活 NOS-NO 通路从而抑制 AT1-R 实现的。
Objective To investigate the effect of Insulin-like growth factor-1 (IGF-1) on the viability and tissue factor (TF) of human umbilical vein endothelial cells (HUVECs) induced by angiotensin Ⅱ Mechanism. Methods HUVEC-12 cells were pretreated with different concentrations of IGF-1 for 30 min and then incubated with Ang Ⅱ at 10 -6 mol / L for 24 h. The changes of cell viability and mRNA expression of AngⅡ type 1 receptor (AT1-R) Quantitative effect relationship. One of the most obvious role of IGF-1 concentration in HUVEC-12 cells pretreated with Ang Ⅱ10 ~ (-6) mol / L incubated for 12 ~ 48 h after 30 min, observed the changes of cell viability of the aging relationship. At the same time, the content of TF, nitric oxide synthase (NOS) and nitric oxide (NO) in HUVEC lysis buffer incubated for 24 h were measured. After pretreatment of cells with NOS inhibitor (L-NAME) for 30 min, -1 and Ang Ⅱ. The changes of cell viability, TF, AT1-R mRNA, NOS and NO levels were observed 24 h after incubation. Results ① Ang Ⅱ could decrease the viability of HUVEC-12 cells, up-regulate the expression of AT1-R mRNA, increase the content of TF, inhibit the activity of NOS and decrease the production of nitric oxide. ②IGF-1 can significantly decrease the cell viability and the expression of AT1- At the concentration of 0.5μg / ml, the effect was most obvious. The cell viability recovery in this concentration-treated cells was time-dependent within 12-48 h. ③IGF-1 could inhibit the increase of TF induced by Ang Ⅱ and the decrease of NOS activity induced by Ang Ⅱ, Generated; ④ L-NAME can significantly antagonize the protective effect of IGF-1 on vascular endothelial cells. Conclusion IGF-1 can significantly increase the activity and antithrombotic ability of endothelial cells induced by Ang Ⅱ, which may be attributed to the inhibition of AT1-R by activating NOS-NO pathway.