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LIU Yun; ZHU Jinquan; SUN Zengxian

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更新时间：2023-03-01

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- China Pharmacy Vol. 27, Issue 34, (2016)
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LIU Yun, ZHU Jinquan, SUN Zengxian. [J]. China Pharmacy 27(34).(2016)
DOI：

LIU Yun, ZHU Jinquan, SUN Zengxian. [J]. China Pharmacy 27(34).(2016) DOI：

摘要

目的：研究转化生长因子β1（TGF-β1）抑制肺动脉平滑肌细胞（PASMCs）凋亡的可能机制。方法：以无血清培养基进行饥饿诱导（SD），MTT法检测0.1、1、5、10 ng/ml的TGF-β1对SD诱导PASMCs细胞存活率的影响。Hoechst 33342染色观察正常对照组、SD组、SD+LY294002[磷脂酰肌醇3-激酶（PI3K）抑制剂]+TGF-β1组、SD+Wortmannin（PI3K抑制剂）+TGF-β1组、SD+TGF-β1组细胞凋亡情况。分别用Western blot法考察正常对照组、SD组、TGF-β1组、SD+TGF-β1组细胞内蛋白激酶B（Akt）、磷酸化（p）-Akt蛋白的表达，正常对照组、SD组、SD+TGF-β1组、SD+LY294002组、SD+LY294002+TGF-β1组细胞环磷腺苷效应元件结合蛋白（CREB）、p-CREB蛋白表达，正常对照组、SD组、SD+TGF-β1组、SD+Wortmannin组、SD+Wortmannin+TGF-β1组细胞CREB和p-CREB蛋白表达，LY294002、Wortmannin、TGF-β1的加入浓度分别为10 μmol/L、50 nmol/L、10 ng/ml。结果：与SD比较，TGF-β1作用后细胞存活率呈剂量依赖性增加，其中5、10 ng/ml质量浓度下细胞存活率差异有统计学意义（P＜0.05）。与正常对照组比较，SD组、SD+LY294002+TGF-β1组、SD+Wortmannin+TGF-β1组细胞凋亡率均增加（P＜0.05）；与SD组比较，SD+TGF-β1组细胞凋亡率下降（P＜0.05）。与正常对照组比较，TGF-β1组细胞内p-Akt蛋白表达增强，p-Akt/Akt增加；与SD组比较，SD+TGF-β1组细胞内p-Akt蛋白表达增强，p-Akt/Akt增加。与正常对照组比较，SD组细胞内CREB蛋白表达减弱；与SD组比较，SD+TGF-β1组细胞内CREB蛋白表达增强；与SD+ TGF-β1组比较，SD+LY294002+TGF-β1组和SD+Wortmannin+TGF-β1组细胞内CREB蛋白表达均减弱，以上差异均有统计学意义（P＜0.05）。结论：TGF-β1可能通过激活PI3K/Akt信号通路抑制PASMCs凋亡。

Abstract

OBJECTIVE： To study the potential mechanism of TGF-β1 inhibiting the apoptosis of pulmonary artery smooth muscle cells （PASMCs）. METHODS： The serum free medium was used for serum deprivation （SD）； MTT assay was used to detect the effects of 0.1， 1， 5， 10 ng/ml TGF-β1 on the survival rate of PASMCs. The apoptosis of PASMCs were observed by Hoechst 33342 staining in normal control group， SD group， SD+LY294002 [PI3K inhibitor]+TGF-β1 group， SD+Wortmannin （PI3K inhibitor）+TGF-β1 group and SD+TGF-β1 group. Western blot technique was applied to investigate the protein expression of Akt and p-Akt in normal control group， SD group， TGF-β1 group and SD+TGF-β1 group； the protein expression of CREB and p-CREB were investigated in normal control group， SD group， SD+TGF-β1 group， SD+LY294002 group and SD+LY294002+TGF-β1 group； the protein expression of CREB and p-CREB were investigated in normal control group， SD group， SD+TGF-β1 group， SD+Wortmannin group and SD+Wortmannin+TGF-β1 group. The concentrations of LY294002， Wortmannin and TGF-β1 were 10 μmol/L， 50 nmol/L， 10 ng/ml， respectively. RESULTS： Compared with SD， survival rate of PASMCs increased as dosage after treated by TGF-β1， with statistical significance at 5， 10 ng/ml （P＜0.05）. Compared with normal control group， apoptotic rate of PASMCs increased in SD group， SD+LY294002+TGF-β1 group and SD+Wortmannin+TGF-β1 group （P＜0.05）； compared with SD group， the apoptotic rate of PASMCs decreased in SD+TGF-β1 group （P＜0.05）. Compared with normal control group， the protein expression of p-Akt and p-Akt/Akt ratio increased in TGF-β1 group； compared with SD group， the protein expression of p-Akt and p-Akt/Akt ratio increased in SD+TGF-β1 group. Compared with normal control group， the protein expression of CREB decreased in SD group； compared with SD group， the protein expression of CREB increased in SD+TGF-β1 group； compared with SD+TGF-β1 group， the protein expression of CREB decreased in SD+LY294002+TGF-β1 group and SD+Wortmannin+TGF-β1 group， with statistical significance （P＜0.05）. CONCLUSIONS： TGF-β1 can inhibit the apoptosis of PASMCs by activating PI3K/Akt signaling pathway.

关键词

肺动脉高压转化生长因子β1磷脂酰肌醇3-激酶/蛋白激酶B凋亡

Keywords

Pulmonary arterial hypertensionTGF-β1PI3K/AktApoptosis

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