Ling-Gui-Zhu-Gan Decoction Protects H9c2 Cells against H(2)O(2)-Induced Oxidative Injury via Regulation of the Nrf2/Keap1/HO-1 Signaling Pathway.
Ling-Gui-Zhu-Gan Decoction Protects H9c2 Cells against H(2)O(2)-Induced Oxidative Injury via Regulation of the Nrf2/Keap1/HO-1 Signaling Pathway.
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DOI：
10.1155/2020/8860603
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发表时间：
2020
期刊：
影响因子：
--
通讯作者：
Zhou P
中科院分区：
文献类型：
--
作者：
Wang X;Tang T;Zhai M;Ge R;Wang L;Huang J;Zhou P
灵归术甘汤是治疗心力衰竭的有效药物，在我们前期的研究中显示出显著的抗炎作用。然而，其通过调节氧化应激反应来改善心力衰竭的能力仍然是未知的。本研究旨在观察灵芝增感汤含药血清对H2 O2诱导的H9 c2细胞氧化损伤的保护作用，并探讨其机制。 将18只大鼠随机分为空白对照组和LGZGD组。LGZGD组大鼠灌胃给予LGZGD 8.4 g/kg/d，连续7 d，空白对照组给予等体积生理盐水。提取所有大鼠的血清。为探讨LGZGD对H9 c2细胞的作用机制，我们将其分为正常对照组、模型组、正常血清对照组、LGZGD组、LGZGD +全反式维甲酸（ATRA）组、ATRA组。丙二醛（MDA）和超氧化物歧化酶（SOD）被用作氧化应激的标志物。采用二氯二氢荧光素二乙酸酯（DCFH-DA）染色法检测细胞内活性氧（ROS）水平。流式细胞仪检测细胞凋亡率。采用蛋白质印迹法检测caspase-3原、切割caspase-3、Bcl-2、Bax、Keap 1、Nrf 2和HO-1的表达水平。使用RT-qPCR测量Keap 1、Nrf 2和HO-1的mRNA水平。 LGZGD可减轻H9 c2细胞的损伤，降低细胞凋亡率。它还能上调SOD活性，抑制MDA和ROS的生成。与模型组相比，LGZGD组caspase-3原、Bcl-2表达水平明显升高，而caspase-3裂解、Bax表达水平明显降低。与模型组相比，LGZGD组Keap 1蛋白表达降低，Nrf 2核表达及Nrf 2介导的转录活性显著升高。ATRA可逆转LGZGD对H2 O2诱导的H9 c2细胞损伤的抗氧化和抗凋亡作用。 我们的研究结果表明，LGZGD减轻H2 O2诱导的H9 c2细胞损伤，通过抑制氧化应激和凋亡Nrf 2/Keap 1/HO-1途径。这些观察结果表明，LGZGD可能通过调节氧化应激反应来预防和治疗心力衰竭。
Ling-Gui-Zhu-Gan decoction (LGZGD) is a potentially effective treatment for heart failure, and it showed significant anti-inflammatory potential in our previous studies. However, its ability to ameliorate heart failure through regulation of oxidative stress response is still unknown. This study was aimed to investigate the protective effect of LGZGD-containing serum on H2O2-induced oxidative injury in H9c2 cells and explore the underlying mechanism. Eighteen rats were randomly divided into two groups: the blank control group and LGZGD group. The LGZGD group rats were administrated with 8.4 g/kg/d LGZGD for seven consecutive days through gavage, while the blank control group rats were given an equal volume of saline. The serum was extracted from all the rats. To investigate the efficacy and the underlying mechanism of LGZGD, we categorized the H9c2 cells into groups: the control group, model group, normal serum control (NSC) group, LGZGD group, LGZGD + all-trans-retinoic acid (ATRA) group, and ATRA group. Malonedialdehyde (MDA) and superoxide dismutase (SOD) were used as markers for oxidative stress. Dichlorodihydrofluorescin diacetate (DCFH-DA) staining was used to measure the levels of reactive oxygen species (ROS). The apoptosis rate was detected using flow cytometry. The expression levels of pro-caspase-3, cleaved-caspase-3, Bcl-2, Bax, Keap1, Nrf2, and HO-1 were measured using western blotting. The mRNA levels of Keap1, Nrf2, and HO-1 were measured using RT-qPCR. The LGZGD attenuated injury to H9c2 cells and reduced the apoptosis rate. It was also found to upregulate the SOD activity and suppress the formation of MDA and ROS. The expression levels of pro-caspase-3 and Bcl-2 were significantly increased, while those of cleaved-caspase-3 and Bax were decreased in the LGZGD group compared with the model group. As compared with the model group, the LGZGD group demonstrated decreased Keap1 protein expression and significantly increased Nrf2 nuclear expression and Nrf2-mediated transcriptional activity. ATRA was found to reverse the LGZGD-mediated antioxidative and antiapoptotic effect on injured H9c2 cells induced by H2O2. Our results demonstrated that LGZGD attenuated the H2O2-induced injury to H9c2 cells by inhibiting oxidative stress and apoptosis via the Nrf2/Keap1/HO-1 pathway. These observations suggest that LGZGD might prevent and treat heart failure through regulation of the oxidative stress response.
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影响因子：
4.5
作者：
Dludla PV;Joubert E;Muller CJF;Louw J;Johnson R
通讯作者：
Johnson R
影响因子：
5.8
作者：
Das, Amlan;Gopalakrishnan, Bhavani;Voss, Oliver H.;Doseff, Andrea I.;Villamena, Frederick A.
通讯作者：
Villamena, Frederick A.
影响因子：
7.4
作者：
Erkens, Ralf;Kramer, Christian M.;Cortese-Krott, Miriam M.
通讯作者：
Cortese-Krott, Miriam M.
影响因子：
7.4
作者：
Canning P;Sorrell FJ;Bullock AN
通讯作者：
Bullock AN
DOI：
10.1007/s00018-016-2223-0
发表时间：
2016-09
期刊：
Cellular and molecular life sciences : CMLS
影响因子：
--
作者：
Loboda A;Damulewicz M;Pyza E;Jozkowicz A;Dulak J
通讯作者：
Dulak J