Hu Shiwen, Wang Pan, Ke Jianhong, Hui Junmin, Wang Cun, Luo Jing, Chen Shaocheng
Chongqing Field Scientific Observation and Research Station for Authentic Traditional Chinese Medicine in the Three Gorges Reservoir Area, Chongqing University of Education, Chongqing 400067, P.R. China.
College of Biological and Chemical Engineering, Chongqing University of Education, Chongqing 400067, P.R. China.
Exp Ther Med. 2023 Sep 21;26(5):517. doi: 10.3892/etm.2023.12216. eCollection 2023 Nov.
Dunn extract (PPDE) is a well-known treatment used in traditional Chinese medicines, where it is most commonly used to treat coughs and symptoms such as headaches and fever. In the present study, the antioxidant capacity of PPDE was determined by scavenging experiments using DPPH, ABTS·, ·OH, and ·O. The cell survival rate was determined by MTT assay. The MDA, SOD, CAT, GSH, and GSH-Px content were determined by colorimetry assays. The expression levels of antioxidant genes SOD, CAT, GSH, and GSH-Px were assessed by reverse transcription-quantitative PCR. HPLC was used to identify the PPDE components. The results suggested that PPDE had scavenging effects on DPPH, ABTS, hydroxyl, and superoxide anion radicals in a concentration-dependent manner; HO treatment resulted in oxidative stress in LLC-PK1 cells, and the degree of injury of LLC-PK1 cells following PPDE treatment was improved, which was positively correlated with its concentration. Dunn extract treatment reduced the content of MDA and increased the content of CAT, SOD1, GSH, and GSH-Px. The mRNA expression levels of antioxidant genes detected by quantitative PCR were consistent with changes in CAT, SOD, GSS, and GSH-Px. Additionally, the trend in CAT, SOD1, GSH, and GSS protein expression levels was also consistent at the mRNA level. PPDE was found to consist of isochlorogenic acid C, myricetin, baicalin, luteolin, and kaempferol. Therefore, PPDE, which was formed of products derived from natural substances, functioned in the inhibition of oxidative damage. The present study aimed to obtain a better understanding of the traditional Chinese medicine Dunn and preliminarily elucidate its antioxidant mechanism at the cellular level. Further animal or human experiments are required to verify the antioxidant effects of PPDE for further development and utilization.
邓恩提取物（PPDE）是一种在传统中药中常用的治疗方法，最常用于治疗咳嗽以及头痛、发烧等症状。在本研究中，通过使用DPPH、ABTS·、·OH和·O进行清除实验来测定PPDE的抗氧化能力。通过MTT法测定细胞存活率。通过比色法测定丙二醛（MDA）、超氧化物歧化酶（SOD）、过氧化氢酶（CAT）、谷胱甘肽（GSH）和谷胱甘肽过氧化物酶（GSH-Px）的含量。通过逆转录定量PCR评估抗氧化基因SOD、CAT、GSH和GSH-Px的表达水平。使用高效液相色谱法（HPLC）鉴定PPDE的成分。结果表明，PPDE对DPPH、ABTS、羟基和超氧阴离子自由基具有清除作用，且呈浓度依赖性；过氧化氢（HO）处理导致LLC-PK1细胞发生氧化应激，PPDE处理后LLC-PK1细胞的损伤程度得到改善，且与浓度呈正相关。邓恩提取物处理降低了MDA的含量，增加了CAT、SOD1、GSH和GSH-Px的含量。通过定量PCR检测的抗氧化基因的mRNA表达水平与CAT、SOD、GSS和GSH-Px的变化一致。此外，CAT、SOD1、GSH和GSS蛋白表达水平的趋势在mRNA水平上也一致。发现PPDE由异绿原酸C、杨梅素、黄芩苷、木犀草素和山奈酚组成。因此，由天然物质衍生的产物形成的PPDE具有抑制氧化损伤的作用。本研究旨在更好地了解传统中药邓恩，并在细胞水平上初步阐明其抗氧化机制。需要进一步的动物或人体实验来验证PPDE的抗氧化作用，以便进一步开发和利用。
