| 谢晓波 潘侨 杨静 刘建伟 唐博恒.辛伐他汀对雷帕霉素作用下心肌微血管内皮细胞的影响[J].,2014,14(21):4033-4037 |
| 辛伐他汀对雷帕霉素作用下心肌微血管内皮细胞的影响 |
| Effect of Simvastatin on Rapamycin-treated Cardiac MicrovascularEndothelial Cells |
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| DOI： |
| 中文关键词: 辛伐他汀 心肌微血管内皮细胞 p70 S6K 一氧化氮合酶 |
| 英文关键词: Simvastatin Cardiac microvascular endothelial cell p70 S6K Reactive oxygen species |
| 基金项目: |
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| 中文摘要: |
| 目的：研究辛伐他汀(SIM)对雷帕霉素(RAPA)引起的体外心肌微血管内皮细胞(CMECs)损害的保护机制。方法：分离、培养
大鼠心肌微血管内皮细胞。经形态学及Dil-ac-LDL吞噬试验进行鉴定后，采用RAPA(100 nM)处理24 小时建立CMECs 损伤模
型，然后加入不同浓度的SIM(0, 10-2，10-1，100，101 uM)培养24小时后，采用MTT,WST-8 及Transwell检测受损后CMECs 的增殖
和迁移；采用Hoechst 33258 及Caspase-3 检查各组CMECs 的凋亡；采用蛋白免疫印迹(Western blotting)检测Akt/p70 S6K 磷酸
化的程度；格里斯反应及实时逆转录PCR 分别检测一氧化氮(NO)含量及一氧化氮合酶(eNOS) mRNA的表达。结果：①经形态学
及Dil-ac-LDL 吞噬试验均表明，成功培养CMECs；②低浓度SIM100 uM 可显著改善100 nMRAPA 对CMECs 增殖、迁移和凋
亡的影响；③SIM 可通过上调PI3K/Akt进而上调p70s6K磷酸化(P<0.05 或P<0.01)；④SIM 通过PI3K/Akt 促进CMECs 分泌NO
及eNOS mRNA 的表达(P<0.05 或P<0.01)。结论：一定浓度下的SIM可改善RAPA 作用下CMECs的增殖，迁移和凋亡，这些保护
作用可能是通过其激活PI3K/Akt/mTOR/p70S6K 信号通路实现的。 |
| 英文摘要: |
| Objective:To investigate the molecular mechanisms underlying the protective effects of simvastatin (SIM) on
rapamycin (RAPA)-treated cardiac microvascular endothelial cells (CMECs).Methods:CMECs isolating from rats left ventricle were
cultured and identified with microscopy and Dil-ac-LDL intake assay. CMECs were then treated with rapamycin (100 nM) for 24 hours.
CMECs treated with 100 nM rapamycin were added with different concentrations of simvastatin (0, 10-2, 10-1, 100, 101 uM) respectively
and then cultured for 24 h. Then cell proliferation and migration were detected by MTT, WST-8 and transwell assays. The apoptosis was
determined by Hoechst 33258 staining and Caspase-3 activity assay. Western blotting was performed to assess the phosphorylation of
Akt/p70S6K. Nitric oxide (NO) secretion was assessed by Griess reaction. The expression of eNOS (endothelial nitric oxide synthase)
mRNA was determined by Real-Time Reverse Transcriptase PCR.Results:① By observing the morphology and Dil-ac-LDL intake
assay indicated that CMECs have cultured successfully. ② Low concentrations of simvastatin (100 uM) significantly improved the
viability, migration and apoptosis on CMECs after pretreated with rapamycin. ③The addition of simvastatin in CMECs resulted in rapid
phosphorylation of Akt, p70 S6 kinase (p70 S6K) (P<0.05 or P<0.01). ④After pretreatment with simvastatin, the expression of NO and
eNOS increased (P<0.05 or P<0.01), which depends the activation of PI3K/Akt/ p70 S6K pathway.Conclusion:Certain concentrations of
simvastatin could improve the viability, migration and apoptosis on CMECs after pretreated with rapamycin. These protective effects may
achieve through the activation PI3K/Akt/mTOR/p70S6K signaling pathway. |
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