Gas6通過PI3K|Akt|FoxO3a通路在預(yù)適應(yīng)中起心肌細胞保護作用
發(fā)布時間：2019-05-12 11:41
【摘要】：目的:初步探討Gas6在預(yù)適應(yīng)中對大鼠心肌細胞缺氧|復(fù)氧損傷的保護作用及機制。 方法:選擇心肌細胞株H9C2建立實驗?zāi)Ｐ?實驗分為5組:①正常對照組:正常培養(yǎng)H9C2細胞;②缺氧復(fù)氧組:缺氧2h,再復(fù)氧24h;③預(yù)適應(yīng)組:先缺氧20min,再復(fù)氧45min,隨后再缺氧2h,再復(fù)氧24h;④加外源性重組人生長停滯特異性蛋白6(rhGas6)組:加入終濃度為100ng/ml的rhGas6,余同缺氧復(fù)氧組;⑤Gas6+LY294002組:加入終濃度為100ng/ml的rhGas6和終濃度為10umol/L的PI3K通路阻斷劑LY294002,余同缺氧復(fù)氧組。應(yīng)用流式細胞術(shù)測細胞凋亡率,測LDH活性了解心肌細胞受損程度;real time -PCR法測Gas6mRNA表達;免疫印跡法測(Western blot)測定磷酸化Akt、FoxO3a蛋白的表達。 結(jié)果:預(yù)適應(yīng)組Gas6 mRNA表達高于缺氧復(fù)氧組(P〈0.05);與缺氧復(fù)氧組對比,預(yù)適應(yīng)組和Gas6組LDH的活性及細胞凋亡率明顯降低(P〈0.05), p-Akt和p-FoxO3a蛋白表達明顯增高(P〈0.05);與Gas6組對比,Gas6+LY294002組LDH的活性及細胞凋亡率明顯增加,p-Akt和p-FoxO3a蛋白表達明顯減少(P〈0.05),而預(yù)適應(yīng)組LDH的活性及細胞凋亡率和p-Akt、p-FoxO3蛋白表達無明顯差異。 結(jié)論:Gas6通過PI3K/Akt/FoxO3a信號通路在預(yù)適應(yīng)中起心肌細胞保護作用。
[Abstract]:Aim: to investigate the protective effect and mechanism of Gas6 on hypoxia / reoxygenation injury in rat cardiomyocytes. Methods: the experimental model of myocardial cell line H9C2 was established and divided into 5 groups: (1) normal control group: normal cultured H9C2 cells, (2) anoxic reoxygenation group: hypoxia for 2 hours, rereoxygenation for 24 hours, and normal control group for 24 hours. (3) preconditioning group: hypoxia for 20 min, then reoxygenation for 45 min, then hypoxia for 2 h, reoxygenation for 24 h and exogenous recombinant human growth stagnation-specific protein 6 (rhGas6) group: the rhGas6, with final concentration of 100ng/ml was added to the hypoxia-reoxygenation group; 5Gas6 LY294002 group: the addition of rhGas6 with final concentration of 100ng/ml and LY294002, a PI3K pathway blocker with final concentration of 10umol/L, was the same as anoxic reoxygenation group. The apoptosis rate was measured by flow cytometry, the activity of LDH was measured to investigate the degree of cardiomyocyte damage, the expression of Gas6mRNA was measured by; real time-PCR method, and the expression of phosphorylated Akt,FoxO3a protein was detected by (Western blot). Results: the expression of Gas6 mRNA in the preconditioning group was higher than that in the hypoxia-reoxygenation group. Compared with hypoxia and reoxygenation group, the activity of LDH and apoptosis rate in pretreatment group and Gas6 group were significantly decreased, and the expression of p-Akt and p-FoxO3a protein was significantly increased (P < 0.05). Compared with Gas6 group, the activity and apoptosis rate of LDH and the expression of p-Akt and p-FoxO3a protein in Gas6 LY294002 group were significantly increased, while the activity, apoptosis rate and apoptosis rate of LDH in preconditioned group were significantly decreased (P < 0.05). There was no significant difference in the expression of p-FoxO3 protein. Conclusion: Gas6 plays a protective role in cardiac myocytes through PI3K/Akt/FoxO3a signaling pathway.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R363
本文編號：2475362
[Abstract]:Aim: to investigate the protective effect and mechanism of Gas6 on hypoxia / reoxygenation injury in rat cardiomyocytes. Methods: the experimental model of myocardial cell line H9C2 was established and divided into 5 groups: (1) normal control group: normal cultured H9C2 cells, (2) anoxic reoxygenation group: hypoxia for 2 hours, rereoxygenation for 24 hours, and normal control group for 24 hours. (3) preconditioning group: hypoxia for 20 min, then reoxygenation for 45 min, then hypoxia for 2 h, reoxygenation for 24 h and exogenous recombinant human growth stagnation-specific protein 6 (rhGas6) group: the rhGas6, with final concentration of 100ng/ml was added to the hypoxia-reoxygenation group; 5Gas6 LY294002 group: the addition of rhGas6 with final concentration of 100ng/ml and LY294002, a PI3K pathway blocker with final concentration of 10umol/L, was the same as anoxic reoxygenation group. The apoptosis rate was measured by flow cytometry, the activity of LDH was measured to investigate the degree of cardiomyocyte damage, the expression of Gas6mRNA was measured by; real time-PCR method, and the expression of phosphorylated Akt,FoxO3a protein was detected by (Western blot). Results: the expression of Gas6 mRNA in the preconditioning group was higher than that in the hypoxia-reoxygenation group. Compared with hypoxia and reoxygenation group, the activity of LDH and apoptosis rate in pretreatment group and Gas6 group were significantly decreased, and the expression of p-Akt and p-FoxO3a protein was significantly increased (P < 0.05). Compared with Gas6 group, the activity and apoptosis rate of LDH and the expression of p-Akt and p-FoxO3a protein in Gas6 LY294002 group were significantly increased, while the activity, apoptosis rate and apoptosis rate of LDH in preconditioned group were significantly decreased (P < 0.05). There was no significant difference in the expression of p-FoxO3 protein. Conclusion: Gas6 plays a protective role in cardiac myocytes through PI3K/Akt/FoxO3a signaling pathway.
【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2011
【分類號】：R363
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相關(guān)期刊論文 前2條
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2 楊彥玲,師養(yǎng)榮,李建龍,陳雅慧;心肌缺血再灌注損傷研究進展[J];心血管病學(xué)進展;2003年02期
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