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红景天苷通过调控PI3K/AKT/Nrf2通路对高糖诱导的<br />心肌细胞的保护作用及机制研究

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摘要:

目的 探究红景天苷(SAL)对高糖诱导的心肌细胞的保护作用并进一步研究其可能的
机制。方法 体外培养H9c2细胞,分为空白组、高糖组、高糖+SAL组、高糖+SAL+LY294002(特异性
PI3K抑制剂)组。空白组细胞用含有5.5 mmoL/L葡萄糖的培养液常规培养;其余三组细胞用含有33.3
mmoL/L葡萄糖的培养液培养;另外高糖+SAL组在更换为高糖培养液前2 h加入29.6 mmol/L SAL预处理,
而高糖+SAL+LY294002组则在加入SAL前加入10 μmol/L LY294002预处理2 h。CCK-8实验和TUNEL实
验检测细胞增殖和凋亡情况。细胞内丙二醛(MDA)、超氧化物歧化酶(SOD)、谷胱甘肽过氧化物
酶(GSH-Px)的含量用相应的酶联免疫试剂盒检测,活性氧簇(ROS)水平通过荧光法检测。蛋白免
疫印迹法(western blot)检测细胞中蛋白激酶b(AKT)、糖原合成酶激酶3β(GSK-3β)、核因子促
红细胞生成素2相关因子2(Nrf2)蛋白表达和磷酸化水平。结果 与空白组相比,高糖组H9c2细胞的存
活率降低,而细胞凋亡率、细胞内ROS合成量和MDA含量增加,细胞内SOD和GSH-Px含量减少,同时
p-AKT/AKT、p-GSK-3β/GSK-3β和Nrf2蛋白表达上调(P均<0.05)。与高糖组相比,高糖+SAL组
H9c2细胞的存活率提高,而细胞凋亡率、细胞内ROS合成量和MDA含量减少,细胞内SOD和GSH-Px含量
增多,且p-AKT/AKT、p-GSK-3β/GSK-3β和Nrf2蛋白表达上调(P<0.05)。与高糖+SAL组相比,高
糖+SAL+LY294002组上述指标均被部分或完全逆转(P<0.05)。结论 SAL对高糖诱导的H9c2细胞凋亡
具有一定的保护作用,可降低细胞内ROS的生成,抑制细胞凋亡,提高细胞存活能力,其作用机制之一
可能与激活PI3K/AKT/Nrf2信号通路有关。

Abstract:

Objective To explore the protective effect of Salidroside (SAL) on cardiomyocytes induced by high
glucose and further study its possible mechanism. Methods H9c2 cells were cultured in vitro and divided into blank
group, high glucose (HG) group, HG+SAL group, HG+SAL+LY294002 group. The cells in the blank group were
cultured in the medium containing 5.5 mmol/L glucose, and the cells in the other three groups were cultured in the
medium containing 33.3 mmol/L glucose. In addition, the HG+SAL group was pretreated with 29.6 mmol/L SAL for
2 hours before changing to HG Medium, while H9c2 cells in the HG+SAL+LY294002 group was pretreated with 10
μmol/L of PI3K inhibitor (LY294002) for 2 hours before adding SAL. CCK-8 method and TUNEL method were used
to detect cell proliferation and apoptosis. The contents of malondialdehyde (MDA), superoxide dismutase (SOD), and
glutathione peroxidase (GSH-Px) were detected by related ELISA kits. Western blot was used to detect the expressions
and phosphorylation levels of protein kinase B(AKT), glycogen synthase Kinase 3-beta (GSK-3β) and nuclear
factor 2 (Nrf2) proteins. Results Compared with the blank group, the survival rate of H9c2 cells in the HG group
were decreased, but the apoptosis rate and the contents of ROS and MDA in H9c2 cells were increased, the contents
of SOD and GSH-Pxin H9c2 cells were decreased. Meanwhile, p-AKT/AKT, p-GSK-3β/GSK-3β and protein
expressions of Nrf2 were up-regulated (P<0.05). Compared with the HG group, the survival rate of H9c2 cells in the
HG+SAL group was increased, but the apoptosis rate and the contents of ROS and MDA in H9c2 cells were decreased,
while the contents of SOD and GSH-Px in the cells were increased, p-AKT/AKT, p-GSK-3β/GSK-3β and protein
expressions of Nrf2 were down-regulated (P<0.05). However, compared with the HG+SAL group, all the above indexes
in the HG+SAL group+LY294002 group were partially or completely reversed (P<0.05). Conclusion SAL can protect
H9c2 cells from high glucose-induced apoptosis, reduce ROS production, inhibit cell apoptosis, and improve cellviability. And one of the mechanisms may be related to the activation of the PI3K/AKT/Nrf2 signalling pathway.

基金项目:

河南省医学科技攻关计划联合共建项目(2018020945)

参考文献:

  • 2008

  • 1

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