左西孟旦对缺氧复氧诱导的H9c2细胞凋亡、PTEN和PI3K/Akt信号通路的影响
作者:
作者单位:

(1.河南科技大学第一附属医院 药剂科,,河南省洛阳市 471000;2.河南科技大学第一附属医院 心血管内科,河南省洛阳市 471000)

作者简介:

李晓玲,副主任药师,研究方向为药学,E-mail:3223731422@qq.com。通信作者汪砚雨,博士,主任医师,研究方向为心血管病,E-mail:13783197120@163.com。

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基金项目:

河南省医学科技攻关计划联合共建项目(LHGJ201905552019)


Effects of levosimendan on hypoxia/reoxygenation-induced apoptosis, PTEN and PI3K/Akt signaling pathway in H9c2 cells
Author:
Affiliation:

1.Department of Pharmacy, Luoyang, Henan 471000, China ;2.Department of Cardiovascular Medicine, the First Affiliated Hospital of Henan University of Science and Technology, Luoyang, Henan 471000, China)

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    摘要:

    目的]研究左西孟旦对缺氧复氧(H/R)条件下H9c2细胞凋亡的影响及相关机制。 [方法]体外培养H9c2细胞,将细胞分为空白对照组、H/R组、左西孟旦低剂量组、左西孟旦中剂量组、左西孟旦高剂量组。采用四甲基偶氮噻唑蓝法检测H9c2细胞增殖;流式细胞仪检测H9c2细胞凋亡;超氧化物歧化酶(SOD)试剂盒、丙二醛(MDA)试剂盒分别检测SOD活性及MDA含量;荧光探针法检测活性氧(ROS)水平;Western blot检测增殖细胞核抗原(PCNA)、B淋巴细胞瘤2蛋白(Bcl-2)、Bcl-2相关X蛋白(Bax)、磷酸酶及张力蛋白同源物(PTEN)及磷脂酰肌醇3激酶(PI3K)/丝氨酸苏氨酸蛋白激酶(Akt)信号通路蛋白表达。 [结果]与空白对照组比较,H/R组H9c2细胞增殖抑制率、凋亡率、MDA含量、ROS水平、PTEN蛋白表达均显著升高(P<0.05),PCNA、Bcl-2、p-PI3K/PI3K、p-Akt/Akt均显著降低,Bax显著升高(P<0.05)。与H/R组比较,左西孟旦低剂量组、左西孟旦中剂量组、左西孟旦高剂量组H9c2细胞增殖抑制率、凋亡率、MDA含量、ROS水平、PTEN蛋白表达均显著降低(P<0.05),PCNA、Bcl-2、p-PI3K/PI3K、p-Akt/Akt均显著升高,Bax显著降低(P<0.05);且左西孟旦剂量越高,上述指标的相应变化越明显。 [结论]左西孟旦可能通过抑制PTEN并激活PI3K/Akt信号通路,促进H/R条件下H9c2细胞增殖,抑制细胞氧化应激和凋亡。

    Abstract:

    Aim To study the effect of levosimendan on apoptosis of H9c2 cells under hypoxia/reoxygenation (H/R) condition and related mechanisms. Methods H9c2 cells were cultured in vitro, and the cells were divided into blank control group, H/R group, low-dose levosimendan group, medium-dose levosimendan group, and high-dose levosimendan group. Proliferation of H9c2 cells was detected by methyl thiazolyl tetrazolium assay; Apoptosis of H9c2 cells was detected by flow cytometry; Superoxide dismutase (SOD) kit and malondialdehyde (MDA) kit were used to detect SOD activity and MDA content respectively; Reactive oxygen species (ROS) level was detected by fluorescent probe method; Proliferating cell nuclear antigen (PCNA), B-cell lymphoma-2 (Bcl-2), Bcl-2 associated X protein (Bax), phosphate and tension homology deleted on chromsome ten (PTEN) and phosphoinositide 3 kinase (PI3K)/serine threonine kinase (Akt) signaling pathway protein expression were detected by Western blot. Results Compared with the blank control group, the proliferation inhibition rate, apoptosis rate, MDA content, ROS level and PTEN protein expression of H9c2 cells in H/R group were significantly increased (P<0.05), PCNA, Bcl-2, p-PI3K/PI3K, p-Akt/Akt were significantly decreased, and Bax was significantly increased (P<0.05). Compared with the H/R group, the proliferation inhibition rate, apoptosis rate, MDA content, ROS level and PTEN protein expression of H9c2 cells in the low-dose levosimendan group, the middle-dose group and the high-dose levosimendan group were significantly decreased (P<0.05), PCNA, Bcl-2, p-PI3K/PI3K, p-Akt/Akt were significantly increased, and Bax was significantly decreased (P<0.05); And the higher the dose of levosimendan, the more obvious the corresponding changes of the above indicators. Conclusion Levosimendan can promote cell proliferation, and inhibit cell oxidative stress and apoptosis in H9c2 cells under H/R conditions possibly by inhibiting PTEN and activating the PI3K/Akt signaling pathway.

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李晓玲,王丹,肖扬,李春燕,汪砚雨.左西孟旦对缺氧复氧诱导的H9c2细胞凋亡、PTEN和PI3K/Akt信号通路的影响[J].中国动脉硬化杂志,2022,30(11):942~948.

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  • 收稿日期:2021-03-04
  • 最后修改日期:2021-07-30
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  • 在线发布日期: 2022-11-07