Kang Seong Hee, Bak Dong-Ho, Lee Seung Sik, Bai Hyoung-Woo, Chung Byung Yeoup, Kang Bo Sun
Research Division for Biotechnology, Advanced Radiation Technology Institute (ARTI), Korea Atomic Energy Research Institute (KAERI), Jeongeup, Jeollabuk 56212, Republic of Korea.
Department of Radiological Science, Konyang University, Daejeon 35365, Republic of Korea.
Exp Ther Med. 2021 May;21(5):419. doi: 10.3892/etm.2021.9863. Epub 2021 Feb 25.
Centipedegrass originates from China and South America, and has been reported to contain several C-glycosyl flavones and phenolic compounds, including maysin and luteolin. The present study aimed to investigate the radioprotective activity of centipedegrass extract (CGE) in radiation exposed-fibroblasts and to assess the affected molecular pathway. The radioprotective effects of CGE were determined in NIH-3T3 cells using Cell Counting Kit-8 and morphological changes were observed. Reactive oxygen species (ROS) levels and the apoptotic profile of NIH-3T3 cells were also measured. The expression levels of B-cell lymphoma-2 (Bcl-2) family proteins [Bcl-2, Bcl-2 like protein 4 (Bax), Bcl-2-associated death promoter (Bad), caspase-3, poly(ADP-ribose) polymerase (PARP)], AKT and MAPK family proteins (ERK, p38 and JNK) were measured . The results demonstrated that when 3T3 fibroblasts pretreated with CGE were subjected to HO-induced cell damage, their viability was significantly decreased. Additionally, CGE pretreatment decreased ROS levels and the protein expression levels of cleaved PARP upon HO treatment, indicating that CGE induced cytoprotective effects against HO-induced oxidative stress. Moreover, significant protective effects of CGE against intracellular ROS, induced upon exposure to ionizing radiation (IR), were observed. The protective effects of CGE pretreatment were also determined by morphological observation of NIH-3T3 cells following exposure to IR. CGE pretreatment increased the expression levels of anti-apoptotic signals (Bcl-2, p-BAD) and decreased the levels of pro-apoptotic signals (Bax, Bad), and led to cleavage of PARP and caspase-3 proteins. Additionally, in cells pretreated with CGE, the phosphorylation of AKT and ERK was increased and that of p38 and JNK was decreased compared with in cells subjected only to IR. These results indicated that CGE may act as a radioprotector due to its anti-oxidative activity, restoring cell homeostasis and redox balance in radiation-exposed fibroblast cells. Therefore, it could be suggested that CGE may be an effective candidate in the treatment of oxidative stress-related diseases and in radioprotection.
假俭草原产于中国和南美洲，据报道含有多种C-糖基黄酮和酚类化合物，包括玉米素和木犀草素。本研究旨在探讨假俭草提取物（CGE）对受辐射成纤维细胞的辐射防护活性，并评估其受影响的分子途径。使用细胞计数试剂盒-8在NIH-3T3细胞中测定CGE的辐射防护作用，并观察形态变化。还测量了NIH-3T3细胞的活性氧（ROS）水平和凋亡情况。检测了B细胞淋巴瘤-2（Bcl-2）家族蛋白[Bcl-2、Bcl-2样蛋白4（Bax）、Bcl-2相关死亡促进因子（Bad）、半胱天冬酶-3、聚（ADP-核糖）聚合酶（PARP）]、AKT和MAPK家族蛋白（ERK、p38和JNK）的表达水平。结果表明，用CGE预处理的3T3成纤维细胞受到HO诱导的细胞损伤时，其活力显著降低。此外，CGE预处理降低了HO处理后ROS水平和裂解PARP的蛋白表达水平，表明CGE对HO诱导的氧化应激具有细胞保护作用。此外，观察到CGE对电离辐射（IR）诱导的细胞内ROS具有显著的保护作用。通过对暴露于IR后的NIH-3T3细胞进行形态学观察，也确定了CGE预处理的保护作用。CGE预处理增加了抗凋亡信号（Bcl-2、p-BAD）的表达水平，降低了促凋亡信号（Bax、Bad）的水平，并导致PARP和半胱天冬酶-3蛋白的裂解。此外，与仅接受IR处理的细胞相比，用CGE预处理的细胞中AKT和ERK的磷酸化增加，p38和JNK的磷酸化降低。这些结果表明，CGE可能因其抗氧化活性而作为一种辐射防护剂，在受辐射的成纤维细胞中恢复细胞内稳态和氧化还原平衡。因此，可以认为CGE可能是治疗氧化应激相关疾病和辐射防护的有效候选物。
