[1]司澳洋,吕风华,尹宏磊,等.抵抗素通过磷脂酰肌醇3激酶/蛋白激酶B信号通路诱导H9c2心肌细胞肥大机制研究[J].新乡医学院学报,2017,34(7):565-569.[doi:10.7683/xxyxyxb.2017.07.003]
　SI Ao-yang,LYU Feng-hua,YIN Hong-lei,et al.Mechanism of resistin induced H9c2 cardiomyocyte hypertrophy by phosphatidyl inositol 3 kinase/protein kinase signaling pathway[J].Journal of Xinxiang Medical University,2017,34(7):565-569.[doi:10.7683/xxyxyxb.2017.07.003]
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抵抗素通过磷脂酰肌醇3激酶/蛋白激酶B信号通路诱导H9c2心肌细胞肥大机制研究
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
34
期数:
2017年7
页码:
565-569
栏目:
基础研究
出版日期:
2017-07-05

文章信息/Info

Title:
Mechanism of resistin induced H9c2 cardiomyocyte hypertrophy by phosphatidyl inositol 3 kinase/protein kinase signaling pathway
作者:
司澳洋; 吕风华; 尹宏磊; 吴　林; 王　卓; 刘亚坤; 张素荣
（新乡医学院第一附属医院心血管内科，河南　卫辉　453100）
Author(s):
SI Ao-yang; LYU Feng-hua; YIN Hong-lei; WU Lin; WANG Zhuo; LIU Ya-kun; ZHANG Su-rong
（1.Department of Vasculocardiology，the First Affiliated Hospital of Xinxiang Medical University，Weihui 453100，Henan Province，China）
关键词:
抵抗素; 磷脂酰肌醇3激酶/蛋白激酶B信号通路; 心肌肥厚
Keywords:
resistin; phosphatidyl inositol 3 kinase/protein kinase B signaling pathway; myocardial hypertrophy
分类号:
R542.2
DOI:
10.7683/xxyxyxb.2017.07.003
文献标志码:
A
摘要:
目的　探讨抵抗素激活磷脂酰肌醇3激酶/蛋白激酶B（PI3K/Akt）信号通路诱导H9c2心肌细胞肥大的分子机制。方法　取生长状态良好的H9c2心肌细胞，使用含1 g·L-1胎牛血清的高糖达尔伯克改良伊格尔培养基（DMEM）处理24 h 使其细胞同步化后进行不同干预。随机将H9c2心肌细胞分为Con组、R50组、LY294002+R50组和LY294002组，Con组为空白对照；R50组细胞应用50 μg·L-1抵抗素处理48 h组；LY294002+R50组细胞预先应用PI3K/Akt通路抑制剂LY294002干预2 h后再用50 μg·L-1抵抗素处理46 h；LY294002组细胞应用PI3K/Akt通路抑制剂LY294002处理2 h组。4组心肌细胞加含药物DMEM培养48 h后进行细胞面积、单细胞蛋白质合成量测定，Western blot检测心肌细胞α-平滑肌肌动蛋白（α-SMA）、原癌基因c-myc蛋白及PI3K/Akt非磷酸化及磷酸化水平表达。观察抵抗素（50 μg·L-1）处理细胞不同时间（0、1、2、3 h）后运用Western blot测定磷酸化（p-Akt）的蛋白表达水平，检测抵抗素处理1 h后4组心肌细胞PI3K/Akt信号通路磷酸化表达水平。 结果　抵抗素处理H9c2心肌细胞0、1、2、3 h组p-Akt蛋白表达水平分别为0.60±0.26、0.71±0.67、0.52±0.32、0.51±0.32；与0 h组比较，抵抗素处理H9c2心肌细胞1 h组p-Akt蛋白表达水平显著增高（P<0.01）；与1 h组比较，抵抗素处理H9c2心肌细胞2、3 h组p-Akt蛋白表达水平显著降低（P<0.01）；抵抗素处理H9c2心肌细胞2 h与处理3 h 后H9c2心肌细胞中p-Akt蛋白表达水平差异无统计学意义（P>0.05）。抵抗素处理H9c2心肌细胞48 h后，与Con组比较，R50组、LY294002+R50组心肌细胞面积、单细胞蛋白含量及α-SMA蛋白表达水平显著升高（P<0.05），而LY294002组其心肌细胞面积、单细胞蛋白含量及α-SMA蛋白表达水平差异无统计学意义（P>0.05）；与R50组比较，LY294002+R50组、LY294002组心肌细胞面积、单细胞蛋白含量及α-SMA蛋白表达水平均显著降低（P<0.05）；与LY294002+R50组比较，LY294002组心肌细胞面积、单细胞蛋白含量及α-SMA蛋白表达水平均显著降低（P<0.05）。Con组、R50组、LY294002+R50组、LY294002组c-myc蛋白表达水平分别为0.45±0.30、0.68±0.32、0.56±0.33、0.16±0.20，4组间比较差异有统计学意义（F=178.91，P<0.05）；与Con组比较，抵抗素处理H9c2心肌细胞48 h后，R50组c-myc蛋白表达水平显著增高（P<0.05）；而LY294002+R50组c-myc蛋白表达水平较Con组、LY294002组升高但低于R50组（P<0.05）。经抵抗素处理H9c2心肌细胞3 h后，R50组PI3K/Akt信号通路磷酸化水平较Con组、LY294002+R50组、LY294002组显著升高（P<0.05），其余各组PI3K/Akt信号通路磷酸化水平表达比较差异无统计学意义（P>0.05）。结论　抵抗素通过激活PI3K/Akt通路，提高Akt磷酸化水平来诱导H9c2心肌细胞肥大。
Abstract:
Objective　To investigate the mechanism of the resistin induced H9c2 myocytes hypertrophy by activating phosphatidyl inositol 3 kinase/ protein kinase B （PI3K/Akt） signal pathway.Methods　The growth in good condition of H9c2 myocytes were obtained and the cells were treated with high glucose Dulkecco modified Eagle medium（DMEM） containing 1 g·L-1 fetal bovine serum for 24 hours to synchronize with different interventions.The H9c2 myocytes were randomly divided into Con group，R50 group，LY294002+R50 group and LY294002 group.The Con group was not given any treatment；the R50 group was treated with resistin （50 μg·L-1） for 48 hours；the LY294002+R50 group was treated with PI3K/Akt pathway inhibitor LY294002 for 2 hours and then treated with resistin （50 μg·L-1） for 46 hours；the LY294002 group was treated with PI3K/Akt pathway inhibitor LY294002 for 2 hours.The cells of the four groups were cultured with DMEM for 48 hours，then the cell area and single cell protein synthesis were measured.Western blot was used to detect the expression of α-smooth muscle actin（α-SMA），oncogene c-myc protein and PI3K/Akt in non-phosphorylation and phosphorylation.Secondly，the level of phosphorylated Akt expression was measured by Western blot at different time （0，1，2，3 h） after treatment with resistin （50 μg·L-1），and then the levels of phosphorylation of the PI3K/Akt signaling pathway in four groups of myocardial cells after the resistance is treated for 1 hour were measured.Results　The phosphorylation levels of Akt protein were 0.60±0.26，0.71±0.67，0.61±0.32 and 0.51±0.32 respectively after H9c2 cardiomyocytes treated for 0，1，2，3 hours with resistin.Compared with the 0 h group，the phosphorylation of Akt protein in H9c2 cells treated with resistin for 1 hour was significantly increased（P<0.01）.Compared with 1 hour group，the phosphorylation level of Akt protein was significantly decreased in 2 hours group and 3 hours group（P<0.01）.There was no significant difference in the phosphorylation of Akt protein between H9c2 cardiomyocytes treated with resistin for 2 hours and 3 hours （P>0.05）.Compared with Con group，the cardiomyocyte area，protein content of single cell and α-SMA protein expression in R50 group，LY294002+R50 group were significantly higher after treated for 48 hours by resistin （P<0.05），and there was no significant difference in myocardial cell area，of single cell protein content and α-SMA protein expression between LY294002 group and the Con group （P>0.05）.Compared with R50 group，the myocardial cell area，of single cell protein content and α-SMA protein expression in LY294002+R50 group and LY294002 group were significantly decreased （P<0.05）.Compared with LY294002+R50 group，the myocardial cell area，of single cell protein content and the expression of α-SMA protein in LY294002 group was significantly decreased（P<0.05）.The expression levels of c-myc protein in Con group，R50 group，LY294002+R50 group and LY294002 group were 0.45±0.30，0.68±0.32，0.56±0.33，0.16±0.20，respectively.There was significant difference in the four groups （F=178.91，P<0.05）.Compared with Con group，the expression of c-myc protein in H9c2 cardiomyocytes in R50 group was significantly higher after treatment by resistin for 48 hours（P<0.05）.The expression of c-myc protein in LY294002+R50 group was significantly higher than that in the Con group and the LY294002 group，but lower than that in the R50 group（P<0.05）.The phosphorylation level of PI3K/Akt signaling pathway in H9c2 cardiomyocytes in the R50 group was significantly higher than that in the Con group，the LY294002+R50 group and the LY294002 group after treated by resistin for 3 hours （P<0.05），and there was no significant difference in phosphorylation level of PI3K/Akt signaling pathway in the Con group，the LY294002+R50 group and the LY294002 group （P>0.05）.Conclusion　The mechanism of resistin induced H9c2 cardiomyocyte hypertrophy involve in activating PI3K/Akt pathway and improving the level of Akt phosphorylation.

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更新日期/Last Update: 2017-07-05