葛根素通过调控PPARγ/NF-κB通路减轻缺氧复氧诱导的心肌细胞损伤

Puerarin mitigates hypoxia-reoxygenation-induced cardiomyocyte injury by modulating the PPARγ/NF-κB pathway

关键词:  葛根素  心肌缺血再灌注损伤  PPARγ/NF-κB  缺氧复氧  细胞凋亡  氧化应激  炎症因子

Key words:  puerarin  myocardial ischemia-reperfusion injury  PPARγ/NF-κB  hypoxia-reoxygenation  apoptosis  oxidative stress  inflammatory mediators

摘要:

[目的] 探讨葛根素（Pue）对缺氧/复氧（H/R）诱导的H9c2心肌细胞损伤的保护作用及机制。[方法] 通过建立H9c2心肌细胞H/R损伤模型（缺氧6 h/复氧2 h），采用不同浓度（25、50、100 μmol/L）的Pue进行干预。通过小干扰RNA（siRNA）技术敲低过氧化物酶体增殖物激活受体γ（PPARγ）表达以验证其机制。采用细胞计数试剂盒-8（CCK-8）法检测细胞活力，蛋白质免疫印迹法（Western blot）检测半胱氨酸-天冬氨酸蛋白酶-3（Caspase-3）、诱导型一氧化氮合酶（iNOS）、PPARγ及磷酸化IκBα（p-IκBα）蛋白表达，酶联免疫吸附法（ELISA）检测炎症因子白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）水平。[结果] H/R处理显著下调PPARγ表达，上调Caspase-3、iNOS和p-IκBα蛋白表达，并增加IL-6和TNF-α的释放。与H/R组相比，Pue组显著上调PPARγ蛋白、抑制核因子κB（NF-κB）通路，同时抑制凋亡、氧化应激相关蛋白及炎症因子的表达。机制研究表明，敲低PPARγ后，Pue的上述保护作用被显著逆转。[结论] Pue通过调控PPARγ/NF-κB信号通路，有效抑制H/R诱导的心肌细胞凋亡、氧化应激与炎症反应，发挥显著的心肌保护作用。

Abstract:

[Objective] To investigate the protective effects and mechanisms of puerarin（Pue） against hypoxia/reoxygenation（H/R）-induced injury in H9c2 cardiomyocytes. [Methods] An H/R injury model was established in H9c2 cardiomyocytes（6h hypoxia/2h reoxygenation），followed by intervention with different concentrations of Pue（25，50，100 μmol/L）. Knockdown the expression of peroxisome proliferator activated receptor gamma（PPARγ） through small interfering RNA（siRNA） technology to validate its mechanism. Cell Counting Kit 8（CCK-8 method） was used to detect cell viability，Western Blot was used to detect the expression of cysteine aspartic acid protease-3（caspase-3），inducible nitric oxide synthase（iNOS），PPARγ and phosphorylated IκBα（p-IκBα） proteins，and enzyme linked immunosorbent assay（ELISA） was used to detect the levels of inflammatory factors interleukin-6（IL-6） and tumor necrosis factor alpha（TNF-α）. [Results] H/R treatment significantly downregulated PPARγ expression，upregulated Caspase-3，iNOS，and p-IκBα protein expression，and increased IL-6 and TNF-α release. Compared with the H/R group，the Pue group significantly upregulated PPARγ protein，inhibited the nuclear factor κB（NF-κB） pathway，and simultaneously suppressed the expression of apoptosis，oxidative stress-related proteins and inflammatory factors. Mechanistic studies revealed that after knocking down PPARγ，the aforementioned protective effects of Pue were significantly reversed. [Conclusion] Pue effectively inhibits H/R-induced cardiomyocyte apoptosis，oxidative stress and inflammatory responses by regulating the PPARγ/NF-κB signaling pathway，exerting significant cardioprotective effects.